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  • 1990-1994  (279)
  • 1830-1839
  • 1990  (279)
  • Genetics  (143)
  • Biochemistry  (136)
  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Journal of insect behavior 3 (1990), S. 579-587 
    ISSN: 1572-8889
    Keywords: Genetics ; polymorphism ; reproductive isolation ; hovering behavior ; Tabanus nigrovittatus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The salt marsh horse fly, Tabanus nigrovittatusMacquart, exhibits two nonoverlapping daily periods of hovering and mating activity, which are correlated with different environmental temperatures. Allelic and genotypic frequencies of hovering males collected during the two periods were compared by electrophoresis of three polymorphic enzyme loci. Approximately 26% of early-hovering males possessed a Pgmallozyme that was absent in our sample of late-hovering males. However, based on other allozyme loci, we found no evidence for reproductive isolation between early and late hoverers. All the genetic data are consistent with the hypothesis that the Pgmpolymorphism is associated with behaviorally and physiologically distinct groups of males that, by all other criteria, form a single Mendelian population.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Pediatric surgery international 5 (1990), S. 359-360 
    ISSN: 1437-9813
    Keywords: Hirschsprung's disease ; Genetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Three male children with identical short-segment Hirschsprung's disease born to a young married couple are reported. There was no positive family history despite an extensive search. There were no associated abnormalities. Although sex-modified multifactorial inheritance, with males having a lower threshold of genes for expression of Hirschsprung's disease, is accepted, the identical expression of the disorder in the three siblings presented suggests a dominant, possibly X-linked gene with variable penetrance. Another possibility is that an identical micro-environmental factor was present prenatally resulting in all three boys having Hirschsprung's disease. This is the first report of three siblings with identical short-segment Hirschsprung's disease.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Pediatric nephrology 4 (1990), S. 533-541 
    ISSN: 1432-198X
    Keywords: Henoch-Schönlein purpura ; Pathogenesis ; Genetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Laboratory studies of the pathophysiology of Henoch-Schönlein purpura (HSP) have become more numerous in recent years with the recognition of the disease's links with the mucosal immune system in general and IgA nephropathy in particular. There are weak genetic associations with C4 null phenotypes and with HLA B35 and DR4. Studies of plasma proteins in HSP patients show an increased IgA concentration, activation of the alternative pathway of complement and consumption of factor XIII. High molecular weight (polymeric) IgA has been detected in affected individuals, which some investigators have called “immune complexes”. Many patients synthesise an IgA rheumatoid factor in the acute phase, but other autoantibodies are largely absent. In vitro studies of lymphocytes from HSP patients have demonstrated an increased number of IgA-bearing and secreting B-cells, with altered T-cell regulation of antibody synthesis. While these observations point to immune dysregulation — primarily of IgA production — as a consistent feature of acute HSP, there is as yet insufficient information available to allow a consistent theory of pathogenesis to be formulated.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Sexual plant reproduction 3 (1990), S. 31-34 
    ISSN: 1432-2145
    Keywords: Mucoraceae ; Zygomycetes ; Homothallic ; Genetics ; Nutritional complementation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Auxotrophic strains of Mucor genevensis and Zygorhynchus exponens were crossed and the resulting zygospores germinated. The presence of a true sexual cycle in both species was demonstrated by the recovery of recombinant genotypes. Expected Mendelian ratios were not realized, however. The presence of selfed zygospores among those isolated makes this observation understandable. It was possible to demonstrate nutritional complementation when young mating mycelium was transferred to minimal medium and forced heterokaryons were recovered.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-1076
    Keywords: Osteogenesis imperfecta ; Collagen type I ; Radiology, classification ; Genetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Congenital osteogenesis imperfecta (OI) was diagnosed by ultrasound in a 31-week-old fetus, and the diagnosis confirmed after delivery by caesarean section at week 36. The baby survived the neonatal period, but failed to thrive, had recurrent respiratory infections and ultimately died at 8 months. Cultured fibroblasts synthesized both normal type I collagen and unstable type I collagen harbouring a structural defect in the α1(I) cyanogen bromide-derived peptide number 8 (CB8) region of the molecule, indicating a heterozygous dominant mutation. A+ birth, the radiological picture was that of the “thin bone”-type of congenital OI (OI type IIB/III in the Sillence classification); at the age of 12 weeks ribs and long bones had undergone a marked expansion giving a very different picture, that of the “thick bone”-type congenital OI (OI type IIA). The mechanism responsible for this change in bone structure is not known, but fractures and callus formation are unlikely to be the only factors. Caution is needed in the interpretation of radiographs of newborns with OI for prognostic or genetic purposes.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Journal of comparative physiology 166 (1990), S. 545-552 
    ISSN: 1432-1351
    Keywords: Honeybees ; Learning ; Classical conditioning ; Selection ; Genetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Four strains of the honeybee (Apis mellifera capensis), which were selected for high (N=2) or low (N=2) performance levels in classic conditioning of olfactory and mechanosensory stimuli, were examined in two instrumental visual learning tasks. Bees were trained to coloured cardboards either at the hive entrance or at the feeding station. Positive correlations were detected between olfactory/mechanosensory conditioning and visual learning. Good and poor learners from strains selected for olfactory conditioning differed significantly in their visual learning values. These strain differences reflect genetic differences in a common learning system rather than task specific differences in sensory, motor or motivational components. Parameters that were influenced by activity of the colony (duration of stay at the feeding place, time between visits) also differed among selected strains. These effects were not due to selection. Instead, they reflect a specific genetic background produced in each strain independently of selection. The results indicate that associative learning has a genetic basis which is independent of the sensory stimuli associated with reward, the learning procedure (classical conditioning or instrumental learning) or the motor patterns used to execute the learned behavior (proboscis extension, control for flight behavior, open field orientation).
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    European archives of psychiatry and clinical neuroscience 239 (1990), S. 290-292 
    ISSN: 1433-8491
    Keywords: Schizophrenia ; Linkage ; Genetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary We analysed six multiplex pedigrees of schizophrenia for linkage to two DNA probes mapping to the chromosome 5q11–q13 region where linkage to a gene for schizophrenia was recently reported. Analyses were conducted using three penetrance models and considering the affected state to be schizophrenia, the schizophrenia spectrum, and all psychiatric diagnoses. All analyses gave consistently negative lod scores. Although the region was not formally excluded, no evidence for linkage was found.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1432-1459
    Keywords: Spinocerebellar degeneration ; Friedreich's disease ; Diagnostic criteria ; Genetics ; Natural history
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The clinical and genetic features of 80 patients with Friedreich's disease from 64 families are described. Diagnostic criteria were: no evidence of dominant inheritance, onset by the age of 20 years, progressive unremitting ataxia of limbs and gait, and absence of knee and ankle jerks. Furthermore, at least one of the following accessory signs was present: dysarthria, extensor plantar response and echocardiographic evidence of hypertrophic cardiomyopathy. Two peaks of onset age were evident at 6–9 and 12–15 years. Analysis of intrafamily variation of onset age and absence of clustering of cardiomyopathy and diabetes did not suggest genetic heterogeneity. Peripheral nerve impairment was an early finding and showed slight further progression, whereas involvement of the cerebellar and corticospinal pathways appeared later and mainly accounted for the progressive worsening of the disease.
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  • 9
    ISSN: 1432-1343
    Keywords: Analysis of variance ; Choropleth map ; Ecology ; Genetics ; Geography ; Permutation test ; Spatial autocorrelation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Mathematics
    Description / Table of Contents: Résumé Cet article présente une solution au problème de l'analyse de variance, pour certains cas où la variable à analyser est spatialement autocorr élée alors que le critère de classification représente des sous-régions connexes du territoire à l'étude. On sait que les méthodes classiques d'analyse de variance ne sont pas applicables dans ce type de situation puisque la condition d'indépendance des échantillons n'est pas respectée; l'autocorrélation positive réduit la variabilité intragroupe, si bien que la quantité relative de variabilité intergroupe s'en trouve artificiellement augmentée. Cette situation correspond en réalité à une vaste catégorie de problèmes en génétique des populations, en écologie et dans d'autres branches de la biologie, ainsi qu'en épidémiologie, en géographie, en géologie, en science économique, en science politique et en sociologie. Ce nouveau test appartient à la famille des tests par permutation. Nous calculons la somme des dispersions intragroupes et testons contre une distribution de référence obtenue en permutant les régions géographiques un grand nombre de fois sur la carte. La véritable difficulté de ce test est d'ordre algorithmique, puisqu'il n'est pas facile de permuter des régions sur une carte, de façon à ce que chaque groupe demeure connexe, et que la carte permutée occupe le même espace total que la carte d'origine. Cet article présente la théorie, les algorithmes, ainsi que des résultats obtenus par cette méthode. Un programme écrit en PASCAL est disponible.
    Notes: Abstract The classical method for analysis of variance of data divided in geographic regions is impaired if the data are spatially autocorrelated within regions, because the condition of independence of the observations is not met. Positive autocorrelation reduces within-group variability, thus artificially increasing the relative amount of among-group variance. Negative autocorrelation may produce the opposite effect. This difficulty can be viewed as a loss of an unknown number of degrees of freedom. Such problems can be found in population genetics, in ecology and in other branches of biology, as well as in economics, epidemiology, geography, geology, marketing, political science, and sociology. A computer-intensive method has been developed to overcome this problem in certain cases. It is based on the computation of pooled within-group sums of squares for sampled permutations of internally connected areas on a map. The paper presents the theory, the algorithms, and results obtained using this method. A computer program, written in PASCAL, is available.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1432-2242
    Keywords: Genetics ; Symbiosis ; Nitrogen fixation ; Coevolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary To determine the relationship between nodulation restriction by the Rj4 allele of soybean, rhizobitoxine-induced chlorosis, and taxonomic grouping of bradyrhizobia, 119 bradyrhizobial isolates were tested in Leonard jar culture for nodulation response and chlorosis induction. In addition to strain USDA 61, the strain originally reported as defining the Rj4 response, eight other isolates (i.e., USDA 62, 83, 94, 238, 252, 259, 260, and 340) were discovered to elicit the nodulation interdiction of the Rj4 allele. Only 16% of all the bradyrhizobial strains tested induced chlorosis, but seven of the nine strains (78%) interdicted by the Rj4 allele were chlorosis-inducing strains. Furthermore, in tests for antibiotic resistance profile, eight of the nine interdicted strains (89%) were classed in DNA homology group II. This evidence suggests that the Rj4 allele has a positive value to the host plant in shielding it from nodulation by certain chlorosis-inducing bradyrhizobia of a DNA homology group with impaired efficiency of nitrogen fixation with soybean.
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  • 11
    ISSN: 1432-2242
    Keywords: Beta vulgaris ; Sugar beet ; Isozymes ; Genetics ; Linkage ; Pollen fertility restorer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The NADP-specific malate dehydrogenase isozymes were controlled by multiple gene systems. Three genes coding for dimeric enzymes segregated in a dependent fashion (NADP-Mdh 1, NADP-Mdh 2, NADP-Mdh 3). A fourth gene (NADP-Mdh 4), also coded for dimers, but was not polymorphic in B. vulgaris. A fifth gene (NADP-Me 1) coded for enzymes active as monomers. Two genes were found to control the main zone of NAD-specific malate dehydrogenase: one coded for dimers (Mdh 1), while a second (Mdh 2) was not polymorphic in the assessions studied. 6-P-Gluconate dehydrogenase was not polymorphic in B. vulgaris; the two types detected on SGE1 electrophoresis were due to developmental expression of the different systems. No genetical segregations could be detected in progeny of crosses of the distinct phenotypes. A shikimate dehydrogenase gene (Skdh 1) that coded for monomers was identified. The diaphorase system was rather complex, but one gene (Dia 1) coding for monomeric enzymes could be identified. Aconitase was found to be controlled by two independent genes (Aco 1, Aco 2), both polymorphic and coding for proteins active as monomers. Tight linkage was found between the genes NADP-Mdh 1, NADP-Mdh 2 and NADP-Mdh 3. Linkage was also found between a pollen fertility restorer (Z) and the Mdh 1 gene. The identification of linkage with Aco 1 needs further investigation. R segregated independently from Mdh 1, Aco 1 and Dia 1. Independent segregations were scored for isozyme genes Pgm 2, Icd 1, Ak 1, Gpi 1, Aco 1 and Dia 1.
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  • 12
    Electronic Resource
    Electronic Resource
    Springer
    Biology and philosophy 5 (1990), S. 349-371 
    ISSN: 1572-8404
    Keywords: Genetics ; gene structure ; hereditary unit
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Philosophy
    Notes: Abstract Definitions of the term ‘gene’ typically superimpose molecular genetics onto Mendelism. What emerges are persistent attempts to regard the gene as a ‘unit’ of structure and/or function, language that creates multiple meanings for the term and fails to acknowledge the diversity of gene architecture. I argue that coherence at the molecular level requires abandonment of the classical unit concept and recognition that a gene is constructed from an assemblage of domains. Hence, a domain set (1) conforms more closely to empirical evidence for genetic organization of DNA regions capable of transcription and (2) has ontological properties lacking in the traditional unit definition.
    Type of Medium: Electronic Resource
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  • 13
    ISSN: 1573-5133
    Keywords: Demography ; Genetics ; Geographic variation ; Stochasticity ; Fluctuating environments ; Allele frequencies ; River ecology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Synopsis The purpose of this study was to determine the effects of unpredictable environmental fluctuations on the demographic and genetic structure of Fundulus zebrinus populations. Collections of F. zebrinus were taken from three rivers in the Arkansas River basin: the Arkansas, Chikaskia, and Ninnescah. Fish were sampled from three sites on each river on nine collection dates throughout 1984 and 1985. Totals of 2100 fish and 6000 fish were included in electrophoretic and demographic analyses, respectively. The results of the study indicate that within a limited geographic region (i.e. within rivers) spatial differences and temporal changes in both demographic and genetic population characteristics occur frequently and are primarily stochastic. However, on a larger spatial scale (i.e. across rivers), general trends emerge for demographic and especially for genetic population characteristics. These results illustrate the importance of sampling scale for conclusions of life-history evolution in fluctuating environments. In addition, it was found that regulation of Fundulus zebrinus populations includes an important density-independent component. Stochastic demographic differences across space and changes through time and spatially and temporally heterogeneous allele frequencies, are both indicative of density-independent regulation. Variation in population parameters, both demographic and genetic, was observed between populations sampled from each river. These population differences were attributed to differences between the rivers themselves.
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  • 14
    Electronic Resource
    Electronic Resource
    Springer
    Journal of chemical ecology 16 (1990), S. 2935-2946 
    ISSN: 1573-1561
    Keywords: Genetics ; sex pheromone ; Lepidoptera ; Noctuidae ; Trichoplusia ni ; cabbage looper moth ; reproductive isolation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The genetic basis of interpopulational differences in the pheromone blend emitted by the cabbage looper moth,Trichoplusia ni (Hübner), was examined by crossing individuals from a field-derived population (P1) with individuals from a long-maintained laboratory colony (P2). These colonies differed in the emission rate and relative proportions of four of the five known minor pheromone components, but not in the emission rate of the major component, (Z)-7-dodecenyl acetate (Z7-12∶Ac). These differences in pheromone blend were quantitatively small but biologically significant, because in the field, males responded preferentially to traps baited with a pheromone blend that is similar to that emitted by P1 females relative to a blend similar to that emitted by P2 females. In initial crosses, variation in the quantity and quality of pheromone blends among families of P1, P2, and F1 hybrid females was examined. In F1 females the relative proportions (quantity relative to the major component) of (Z)-5-dodecenyl acetate (Z5-12∶Ac) and (Z)-7-tetradecenyl acetate (Z7-14∶Ac) were intermediate to parental lines. In a second more extensive set of crosses, analyses included P1, P2, F1, F2, and selected backcrosses. The relative proportion of Z5-12∶Ac, Z7-14∶Ac, and Z9-14∶Ac emitted by F1 females were intermediate to parental lines. The frequency distributions of relative proportions of these components emitted by females were not consistent with those expected under a single autosomal or sex-linked gene hypothesis, suggesting that more than one gene is involved in the quantitative differences in the pheromone blend.
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  • 15
    Electronic Resource
    Electronic Resource
    Springer
    Journal of comparative physiology 160 (1990), S. 183-186 
    ISSN: 1432-136X
    Keywords: Histology ; Biochemistry ; Muscle physiology ; Hibernations ; Metabolism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Fiber composition, and glycolytic and oxidative capacities of the pectoralis, gastrocnemius, and cardiac muscles from active and hibernating little brown bats (Myotis lucifugus) was studied. The data were used to test two hypotheses: First, since hibernating bats maintain the capability of flight and make use of leg muscles to maintain a roosting position all winter, the fiber composition of the pectoralis and gastrocnemius muscles should not change with season. Second, we tested the hypothesis of Ianuzzo et al. (in press), who propose that the oxidative potential of mammalian cardiac muscle should increase with increasing heart rate while glycolytic potential should not. Our results indicate that the fiber composition of the pectoralis muscle was uniformly fast-twitch oxidative (FO)_ regardless of the time of year, as predicted. However, the gastrocnemius muscle exhibited a change in FO composition from 83% in active to 61% in hibernating animals. Contrary to the variable change in histochemical properties with metabolic state, a trend of reduced maximal oxidative (CS) and glycolytic (PFK) potential during hibernation in both flight and leg muscles was apparent. The oxidative potential of flight and leg muscles decreased by 15.2% and 56.5%, respectively, while the glycolytic potential of the same muscles decreased by 23.5% and 60.5%, respectively. As predicted, the glycolytic potential of cardiac muscle remained constant between active and hibernating bats, although there was a significant decrease (22.0%) in oxidative potential during hibernation.
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  • 16
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 27 (1990), S. 168-172 
    ISSN: 1040-452X
    Keywords: Motility ; Genetics ; Sex chromosome ratio ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In this study, we address the relationship between motility and genetic content of mouse sperm. The chromosome complements of highly motile mouse sperm, selected using the swim-up technique, were analyzed after in vitro fertilization, at the first cleavage state. They were compared to those of unselected sperm. Identification of male and female chromosome sets was possible because of their differential condensation at the first mitotic division. In vitro fertilization, swim-up separation, chromosome preparation, and staining were carried out using standard techniques. The results indicate that highly motile mouse sperm did not differ in types and frequencies of chromosomal abnormalities from those not selected for motility. Moreover, separation of motile sperm does not deviate the sex ratio from the theoretical 1:1.
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  • 17
    Electronic Resource
    Electronic Resource
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 88-96 
    ISSN: 0192-253X
    Keywords: Pattern formation ; segmentation ; gap genes ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: L(1)giant is a zygotic lethal mutation which affects the embryonic development of both the labial/thoracic segments and a subset of posterior abdominal segments. Using antibodies specific for proteins encoded by several Drosophila genes to identify the compartmental origin of the defects, we show that the requirement of giant activity is different in these two embryonic domains. Anteriorly, the posterior compartment of the labial segment is missing at the blastoderm stage. Posteriorly, cells are specifically deleted by cell death within the anterior compartments of abdominal segments 5-7 during germ band elongation. In mature embryos, posterior compartment structures of the peripheral nervous system of A5-7 are fused. In addition to a different pattern of defect in the two parts of the embryo, the kind of action appears different. Anteriorly, giant resembles a gap mutation in that a particular region is missing from the blastoderm fate map, whereas in the abdominal domain, giant affects the development of anterior compartment-specific structures.
    Additional Material: 4 Ill.
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  • 18
    Electronic Resource
    Electronic Resource
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 123-123 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 19
    Electronic Resource
    Electronic Resource
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990) 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 20
    ISSN: 0192-253X
    Keywords: Heat shock promoters ; HSP70-CAT ; microinjection ; linker-scanner mutations ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Microinjected human HSP70 promoter-chloramphenicol acetyl transferase (CAT) chimeric genes are constitutively expressed immediately after the midblastula transition of Xenopus embryogenesis. Analysis of a series of 5′-deletion mutants in the HSP70 promoter revealed that sequences within 74 bases of the transcriptional start site were sufficient for strong basal activity. We investigated the role of specific sequences in the basal promoter by injecting HSP70-CAT vectors containing linker-scanner mutations in the basal elements (CCAAT, purine-rich element, GC-element, ATF/AP1, and 1ATA). Our data reveal that deletion of any of these cis-acting elements in the basal promoter prevents expression after the midblastula stage of development. Furthermore, we have identified specific binding activities in embryonic nuclear extracts that complex with basal promoter elements (CCAAT, ATF, and GC) of the heterologous HSP70 promoter. These trans-acting factors are detectable in nuclear extracts of early blastula embryos, and their respective binding activity increases dramatically after the midblastula transition. The expression of the human HSP70 gene after the midblastula transition of Xenopus embryogenesis requires an array of cisacting elements, which interact with specific Xenopus transcription factors.
    Additional Material: 9 Ill.
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  • 21
    Electronic Resource
    Electronic Resource
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 110-122 
    ISSN: 0192-253X
    Keywords: Embryonic cell surface ; glycoconjugates ; monoclonal antibodies ; developmental expression of glycoconjugates ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Molecular markers for specific cell lineages would be useful in studies of cellular differentiation. To isolate such markers monoclonal antibodies (MoABs) were raised against plasma membranes isolated from gastrulating Xenopus embryos. Those antibodies that recognized subsets of cells within the embryo were selected by indirect immunofluorescence. The analysis of eight such MoAbs is presented. Western blot analysis showed that all but one MoAb recognized a complex pattern of glycoconjugates associated with glycoproteins. All the antigens recognized by the MoAbs were maternal in origin and displayed similar spatial patterns of pregastrular expression. This pattern of immunoreactivity at the apical surface was inherited passively during cleavage by the resulting superficial blastomeres suggesting that ectodermal specific markers of maternal origin are pre-localized to the cortical ooplasm in mature oocytes. We suggest that these maternal components may be specific glycosyl transferases. Three different patterns of expression were observed during gastrulation as exemplified by MoAbs 1F10C1, 3A4D1, and 6F10B6. MoAb 6F10B6 was specific for both neural and non-neural epithelium. MoAb 3A4D1 was specific for non-neural epidermis. MoAb 1F10C1 appeared to recognize a protein epitope on an extracellular component expressed by the superificial and involuting epithelial cells. The pattern of expression for the 1F10C1 antigen suggests that it may play a role in facilitating the movement of the involuting cells during gastrulation.
    Additional Material: 7 Ill.
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  • 22
    Electronic Resource
    Electronic Resource
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 125-132 
    ISSN: 0192-253X
    Keywords: Flow cytometry ; polytenization ; C value ; fluorescence ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Multiparametric flow cytometry was used to analyze the development of the endosperm in Zea mays L. during the period from 8 to 20 days after pollination (dap). Nuclear size, DNA content per nucleus, and frequencies of nuclei with varying properties were measured in preparations that included all of the endosperm nuclei of single kernels of the inbred strain Al88. Characteristics of nuclear populations from different kernels on the same ear showed minimal variation. The dynamic changes of non-mitotic cells involved in endosperm development consisted of alternating periods of DNA replication with non-replication. Seven rounds of DNA replication had occurred in some nuclei in the later developmental stages with the rate averaging approximately one round per 24-hour period. Analysis of the DNA levels in the nuclei showed an exact doubling pattern indicating an endoreduplication process, that is, replication of the entire genome during each round. The loosely organized polytenization of the chromatin occurred to varying extents among the nuclei within an endosperm. A weak positive correlation existed between DNA content and size of nuclei suggesting that DNA increases and nuclear growth may not be highly coordinated in this tissue. Increased proportions of the larger nuclei occurred in the later stages of endosperm development. Considering the entire endosperm, the average DNA content per nucleus at the 15-dap peak level was approximately 12.8 C constituting a 2.7-fold overall increase from 8 dap.
    Additional Material: 8 Ill.
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  • 23
    Electronic Resource
    Electronic Resource
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 176-181 
    ISSN: 0192-253X
    Keywords: Direct gene transfer ; transgenic plants ; expression of transgenes ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Direct gene transfer to protoplasts is one of several methods developed for the production of transgenic plants. This method utilizes the efficient uptake of DNA from the surrounding medium by protoplasts (cell wall-less plant cells). Where a suitable protoplast system exists large numbers of transformant clones can be efficiently produced and often regenerated to normal fertile plants. This review concentrates on the fate of the DNA which is taken up into the protoplasts. Particular emphasis is given to the factors which can influence the integration and form of the transferred DNA, the expression of transferred genes, and the inheritance in further generations of those genes. The information available suggests (1) that DNA is taken up by a large proportion of the cells in a transformation mixture, (2) that this DNA forms complexes sometimes involving carrier DNA, (3) that fewer cells actually take up DNA into the nucleus, and (4) that the complex may be rearranged and/or amplified and then integrated into the genome. If the DNA is arranged in such a way that a gene can be expressed it does so in a normal manner and is stably inherited both mitotically and meiotically.
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  • 24
    Electronic Resource
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 197-204 
    ISSN: 0192-253X
    Keywords: Light-regulated genes ; transgenic plants ; enhancer ; silencer ; regulatory elements ; trans-acting factors ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Transgenic plants have been particularly useful in studying nuclear genes encoding for photosynthetic functions. The expression of these genes and their chimeric constructs in transgenic plants faithfully mimics their natural counterparts. The use of sensitive chimeric reporter genes has enabled localizing the activity of genes encoding photosynthetic proteins to individual cells. Cab and rbcS transgenes have been shown to retain sensitivity to light quality, which is modulated by phytochrome. Conditional light activation under the influence of a circadian rhythm has been shown for Cab transgenes. Transgenic plants containing truncated promoters have helped delineate cis-regulatory positive and negative elements involved in light-mediated transcriptional induction and tissue specificity.
    Additional Material: 1 Ill.
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  • 25
    ISSN: 0192-253X
    Keywords: nodulins ; leghemoglobin ; glutamine synthetase ; Enod2 ; cis-acting elements ; transacting factors ; Agrobacterium turnefaciens ; A. rhizogenes ; binary vectors ; plant transformation ; chimeric genes ; chloramphenicol acetyltransferase ; glucuronidase ; cytokinin induction ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Nodulin genes are plant genes specifically activated during the formation of nitrogen-fixing nodules on leguminous plants. These genes are interesting to study since they are not only induced in a specific developmental fashion by signals coming directly or indirectly from the rhizobial symbiont, but are also expressed in a tissue-specific manner. By examining the expression of chimeric nodulin-reporter genes in transgenic legume plants it has been shown that nodule specific expression is mediated by DNA sequences present in the 5′upstream region of several nodulin genes. Here we summarize the available data on these cis-acting elements and the trans-acting factors interacting with them. We also review experiments designed to identify rhizobial “signals” which may play a role in nodule specific gene expression.
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  • 26
    Electronic Resource
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 205-213 
    ISSN: 0192-253X
    Keywords: cauliflower mosaic virus 35S RNA ; cell cycle gene expression ; cis-acting element ; wheat histone H3 gene ; trnas-acting factor ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A majority of histone genes are expressed in the S phase during the cell cycle. Using the gene expression system of transformed sunflower cells into which wheat histone H3 gene was introduced by the Ti-plasmid gene transfer technique, we determined three cis-acting control sequences (hexameric, octameric, and nonameric motifs) which seemed to confer the S-phase-specific transcription of wheat histone genes. Furthermore, as candidates for regulatory transcription factors, three nuclear DNA-binding proteins HBP-la, HBP-lb, and HBP-2 that interact with the hexameric and nonameric motifs were identified. The structural analysis of the cDNA of HBP-la revealed that a nuclear protein has the leucine-zipper structure and a DNA-binding motif. The hexameric motif in the H3 gene was also seen in cauliflower mosaic virus 35S (CaMV 35S) promoter and shown to function as a regulatory element of this promoter. The wheat HBP-1b can interact with the hexameric motif of the CaMV 35S promoter. Much attention has been paid to the significance of the hexameric sequences within the H3 and CaMV 35S promoters and the DNA-binding proteins HBP-la and HBP-lb.
    Additional Material: 4 Ill.
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  • 27
    Electronic Resource
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990) 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 28
    ISSN: 0192-253X
    Keywords: lux ; luc reporter genes ; light emission ; gene expression ; single photon imaging in vivo ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Genes encoding light-emitting luciferase were recently isolated from luminous marine bacteria and fireflies. Expression of luciferase genes in diverse organisms is a unique way for studying gene expression by simple and sensitive measurement of light. Recent advances in application of luciferase reporter genes are reviewed and documented by examples of in vivo visualization of their expression in transgenic plants.
    Additional Material: 2 Ill.
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  • 29
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 214-223 
    ISSN: 0192-253X
    Keywords: Gene interactions ; cytosine methylation ; epigenetic variation ; transgenic plants ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Unusual gene interactions were observed in several doubly transformed tobacco plants which were obtained following sequential transformation steps using two T-DNAs encoding different selection and screening markers. The expression of T-DNA-I, which encoded kanamycin resistance (Kanr) and nopaline synthase (NOS), was suppressed in some, but not all, of the double transformants after the introduction of T-DNA-II, which encoded hygromycin resistance (Hygr) and octopine synthase (OCS). Double transformants in which T-DNA-I had been inactivated could produce KanrNOS+ progeny, but these were shown to lack T-DNA-II, thus establishing the role of this T-DNA in the suppression of T-DNA-I. Reversible cytosine methylation of the promoters of T-DNA-I genes was shown to correlate with their activation/inactivation cycle. In this paper we pursue further the questions of the mechanism of suppression of T-DNA-I genes by T-DNA-II, and also the timing and extent of demethylation of T-DNA-I promoters in Kanr progeny following the loss of T-DNA-II. We propose that the suppression is due to the competition between homologous regions on each T-DNA for binding to nuclear sites with fixed locations. We further suggest that incomplete demethylation patterns of T-DNA-I promoters in Kanr progeny reflect the existence in the shoot apex meristem of two cell populations, which have either methylated or unmethylated T-DNA-I promoters, respectively. Thus, Kanr progeny are epigenetic chimeras with respect to the expression of T-DNA-I genes.
    Additional Material: 7 Ill.
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  • 30
    Electronic Resource
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 233-247 
    ISSN: 0192-253X
    Keywords: Chromatin structure ; foreign gene expression ; transgenic plants ; Nicotiana tabacum ; position effect ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The chromatin structure of foreign genes in transgenic tobacco plants was investigated by digestion of nuclei with DNase I and micrococcal nuclease, respectively, followed by restriction and Southern analysis of the digestion products. The results were compared to the differential expression of the different transgenes. Two model systems were used: plants harbouring vector DNA derived from the disarmed vector pGV 3850 and plants harbouring the light-regulated and organ-specifically expressed potato ST-LS1 gene and the cotransferred ncpaline synthase (nos) reporter gene. Our results show that transferred genes are located in DNase l-sensitive domains in all transformants. Slight variations of DNase l-sensitivity of the transferred ST-LS1 constructs in different transformants neither reflected the between-transformant variability of expression nor the organ-specific activity of the transgenes. A deletion event was found responsible for silencing the ST-LS1 gene but not the nos gene in one of the transformants. Whereas no DNase l-hypersensitive sites were found within the 3850-T-DNA and the ST-LS1 gene, one prominent site was mapped to the nos promoter within the ST-LS1 construct in all transformants. Digestion of chromatin harbouring 3850-T-DNA with micrococcal nuclease resulted in a blurred nucleosomal pattern as compared to nucleolar and bulk chromatin, the extent of blurring being independent of the expression of transferred genes. The present results favour the “permissive domain” hypothesis which capitalizes on the chromatin surrounding the integration site as the determining factor for the chromatin structure of incoming alien genes. However, between-transformant variability of expression is not reflected by differential sensitivity to DNase I. Hence, other factors than chromatin structure must be involved in creating “position effects”.
    Additional Material: 9 Ill.
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  • 31
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 249-253 
    ISSN: 0192-253X
    Keywords: Drosophila ; embryonic development ; sex differentiation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We report the first practical application of a genetic scheme devised for the purpose of obtaining large quantities of embryos of a specific sex. The scheme, which is based on the meiotic drive system Segregation Distorter, results in the production of populations of zygotes that are almost exclusively of one sex. We have used this scheme to determine that the steady-state levels of transcripts of X-linked genes are the same in early male and female embryos, establishing that these genes are dosage compensated.
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  • 32
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 263-269 
    ISSN: 0192-253X
    Keywords: Penetrance ; reciprocal effects ; suppressor genes ; developmental cycle in mammals ; initialization ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The phenomenon of gametic imprinting in mammals has raised developmentally relevant questions concerning the manifestation and inheritance of genes with variable penetrance. The dominant fused (Fu) gene located on chromosome 17 is one of the few good cases demonstrating the phenomenon in mice. The Fu mutation has a maternal effect.We have previously shown that the † 12 haplotype significantly lowers the penetrance of Fuin ♀ ♀ Fu/†12 offspring. Results of recipiocal matings of the heterozygotes for Fu indicated that the Fu of maternal origin has a lowered level of penetrance. The dominant suppressors locotad outside chromosome 17, in contrast to †12 residing in it, had stronger effects on the manifestation of Fu, decreasing its penetrance to 8-17%. Experimental evidence is presented that the pathway via which Fu passes to the zygote nucleus during gametogenesis through successive generations has a marked effect on its penetrance. Based on this evidence, patterns of genetic imprinting are described. A survey of genetic imprinting allowed us to distinguish two developmental phases, gametic and zygotic. The hypothesis for the gametic phase of the development of multicellular organisms suggests that it proceeds from initialization, a process thought to ensure the freeing of chromosomes from redundant epigenetic information and their preparation for the consecutive developmental cycle.
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  • 33
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 318-325 
    ISSN: 0192-253X
    Keywords: Mice ; housekeeping genes ; liver ; tissue specificity ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In situ hybridization using nucleic acid probes was used to detect cell- and tissue-specific transcript(s) of embryonic genes during development and differentiation. This highly sensitive technique has the potential to provide valuable information on the regulation of low-abundance housekeeping genes during development. We have determined the experimental conditions required to detect the catalase message in adult mouse liver. Catalase effects the breakdown of H2O2 to O2 and H2O and offers protection against the toxic effects of oxygen radicals. We used a cloned 550 bp BamHI-Pstl fragment from a mouse catalase cDNA (pMCT-1) to generate 35S-labeled sense and antisense riboprobes. The experimental conditions used were sensitive enough to quantitate the abundance of silver grains generated by the antisense riboprobe on the adult liver, a tissue known to be positive for this message. The hybridization protocol was applied to serial sections of 13- and 18-day-old mouse embryos. The results suggest that the catalase expression in the liver and brain begins with somite formation and increases with development and differentiation. On the other hand, this message appears to be absent in mesenchyme, particularly in day 13 embryos. The message in positive tissues appears evenly distributed throughout the cell. The observed expression of the catalase message in the adult liver is approximately six times that in the embryonic liver. It is compatible with the enzyme activity results and emphasizes the sensitivity of the in situ hybridization method (over northern blot, etc.) used in this study.
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  • 34
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 328-332 
    ISSN: 0192-253X
    Keywords: Cellular slime molds ; cytoskeleton ; actin-binding proteins ; review ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The cellular slime mold Dictyostelium discoideum is becoming the premier system for the explication of the biochemical and cellular events that occur during motile processes. Proteins associated with the actin cytoskeleton, in particular, appear to play key roles in cellular responses to many external stimuli. This review summarizes our present understanding of the actin-associated proteins in Dictyostelium, including their in vitro activities and their structural and/or functional analogues in mammalian cells.
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  • 35
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 333-340 
    ISSN: 0192-253X
    Keywords: Dictyostelium discoideum ; actin ; ABP-120 ; elongation factor 1 alphc ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In this work we evaluate the cartical expansion model for amoeboid chemo-taxis with regard to new information about molecular events in the cytoskeleton following chemo-tactic stimulation of Dictyostelium amoebae. A rapid upshift in the concentration of chemoattrac-tant can be used to synchronize the motile behavior of a large population of cells. This synchrony presents an opportunity to study the biochemical basis of morphological changes such as pseudopod extension that are required for amoeboid chemotaxis. Changes in the composition and activity of the cytoskeleton following stimulation can be measured with precision and correlated with important morphological changes. Such studies demonstrate that activation of actin nucleation is one of the first and most crucial events in the actin cytoskeleton following stimulation. This activation is followed by incorporation of specific actin cross-linking proteins into the cytoskeleton, which are implicated in the extension of pseudopods and filopods. These results, as well as those from studies with mutants deficient in myosin, indicate that cortical expansion, driven by focal actin polymerization, cross-linking and gel osmotic swelling, is an important force for pseudopod extension.It is concluded that whereas three forces, frontal sliding, tail contraction, and cortical expansion may cooperate to produce amoeboid movement, the cortical expansion model offers the simplest explanation of how focal stimulation with a chemoat-tractant causes polarized pseudopod extension.
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  • 36
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 362-368 
    ISSN: 0192-253X
    Keywords: Dictyostelium ; filopodia ; actin binding proteins ; calcium ; cell motility ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The Dictyostelium discoideum 30,000 dalton actin-binding protein is an actin cross-linking protein that organizes formation of parallel bundles of actin filaments in vitro, and is present in filopodia in living cells. This protein binds calcium directly and exhibits a decreased affinity for actin filaments in the presence of micromolar calcium. In this work, the existence of antigenic homologs of the 30,000 dalton protein in Physarum polycephalum, Schistosoma mansoni, Chara carolina, and Drosophila melanogaster is detected by use of affinity purified antibody and electrophoretic blotting methods. The expression of this protein during development of Dictyostelium is also analyzed, revealing a progressive 3-fold decrease in the level of this protein in amoebae between the vegetative and slug stages. A highly ordered structure of bundles of actin and the 30,000 dalton protein formed in vitro is inferred from the presence of transverse striations on the bundles with a minor periodicity at 11.4 nm and a major periodicity at 33.9 nm. Finally, we propose a working model of the interaction of this actin cross-linking protein with actin filaments to form bundles.
    Additional Material: 5 Ill.
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  • 37
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 369-376 
    ISSN: 0192-253X
    Keywords: Cytoskeleton ; capping proteins ; genamic structure ; Dictyostelium ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The two subunits of the het-erodimeric protein cop32/34, an actin-binding protein, are encoded by separate single-copy genes. We have established the genomic structure of both genes. A sequence comparison of cap32/34 with capZ from chicken skeletal muscle and two partially known sequences from Saccharomyces cerevisiae and Xenopus laevis show that heterodimeric capping proteins belong to a highly conserved group of actin-binding proteins. This conclusion is supported by the cross-reaction of polyclonal antibodies against cap32 and cap34 with proteins from lower and higher eukaryotes. In addition, a system is presented that allows the expression of truncated cap34 polypeptides under the control of the cap34 promoter.
    Additional Material: 5 Ill.
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  • 38
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 388-390 
    ISSN: 0192-253X
    Keywords: Transformation ; electroporotion ; gene targetting ; antisense RNA ; complementation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: DNA mediated transformation is a critical technique for uniting genetic, molecular genetic and reverse genetic approaches to a wide range of problems in cell and molecular biology. In Dictyostelium, there is now the capability not only to manipulate DNA sequences in vitro and put them back into the cell, but also to alter the sequences of endogenous chromosomal genes through high frequency homologous recombination. This means that the range of gene manipulation techniques that have made yeast such a useful system should now be applicable in Dictyostelium. For studying problems such as cellular motility, morphogenesis, and gene regulation, few organisms have the combination of features offered by Dictyostelium.
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  • 39
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 391-395 
    ISSN: 0192-253X
    Keywords: Lipofectin ; slime mould ; transformation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have studied the transient expression, in Dictyostelium cells growing on a bacterial food source, of a construct containing the coding region of the firefly luciferase gene inserted downstream of a Dictyosteliumactin promoter. The fusion gene is not detectably expressed when DNA is introduced by calcium phosphate precipitation or by electroporation, but it is expressed when introduced using cationic liposomes (lipofectin). Using this latter procedure, we are able to transform cells with a G418 resistance vector and select stable, drug-resistant transformants at a relatively low, but workable, efficiency. This technique will allow molecular genetics to be applied to the many important nonaxenic Dictyostelium strains.
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  • 40
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 377-387 
    ISSN: 0192-253X
    Keywords: Cell-cell ; adhesion molecule ; cell binding site ; cell aggregation ; development ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: During development of Dictyostelium discoideum, cells acquire EDTA-resistant cell-cell adhesion at the aggregation stage. The EDTA-resistant cell binding activity is associated with a cell surface glycoprotein of Mr 80,000 (gp80), which mediates cell-cell binding via ho-mophilic interaction. Analysis of the structure of gp80 deduced from cDNA sequence reveals the presence of three internally homologous segments in the NH2-terminal domain, which also contains regions with homology to the neural cell adhesion molecule. Secondary structure predictions show an abundance of β-structures and very few α-helices. This is confirmed by circular dichroism measurements. It is likely that the homologous segments are organized into globular structures, extended from the cell surface by a Pro-rich stalk domain. The cell binding activity of gp80 resides within the first globular repeat of the NH2-terminal domain and has been mapped to a 51 amino acid region between Val123 and Leu 173. Synthetic oligopeptides corresponding to sequences within this region have been prepared and assayed for their ability to bind to cell surface gp80. Results lead to identification of the homophilic binding site to an octapeptide sequence within this region. Synthetic peptides containing this octapeptide sequence and univalent antibodies directed against this site block the formation of organized cell streams during aggregation. Although cell aggregates are eventually formed, most fail to undergo further development to give rise to slugs and fruiting bodies, indicating that cell-cell adhesion involving gp80 is an important step in normal morphogenesis.
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  • 41
    ISSN: 0192-253X
    Keywords: Transformation ; Dictyostelium discoideum ; UMP-synthase/ura-complementation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Whereas transformation ofDictyostelium discoideum now can be done routinely and reliably, there is increasing demand for a system that allows selection of cells that have gone through repeated transformation cycles. Such a system is presented here. Selection is based on resistance to 5-fluoro-orotic-acid (5-FOA). D. discoi-deum is highly sensitive to this drug, and cells can survive only in the presence of 5-FOA if they carry a defective UMP-synthase gene. After transformation with a plasmid carrying a cloned version of the UMP-synthase gene, 5-FOA resistant cells are obtained at high frequency. Because 5-FOA-resistant cells depend on exogenously added uracil, these cells can be taken through a second round of transformation. A plasmid-borne UMP-synthase gene renders 5-FOA-resistant cells phenotypically ura +. Finally, cells can be further transformed using the standard G418 selection system.
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  • 42
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 425-426 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 43
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 403-409 
    ISSN: 0192-253X
    Keywords: Electroporation ; soft-agarose ; G418 ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A new method for clonal growth of Dictyostlium axenic amoebae has been developed. Cells are plated in growth medium containing 1% ultra-low gelling temperature agarose. Cells grow normally in the agarose and form colonies up to several millimeters in diameter. When the colonies have grown to a sufficient size, they begin multicellular development. Pseudoplasmodia are formed, migrate to the surface of the agar, and then undergo fruiting body formation. Cells can be removed from the soft agarose colonies with a toothpick or by picking spores from the fruiting bodies. This method should be useful for drug, auxotrophic, and temperature selections where clonal maintenance of axenic colonies is important.This method has been used in combination with a selection for resistance to G418 to isolate independent colonies following DNA-mediated transformation. Several parameters in the calcium phosphate and electroporation transformation protocols have been optimized and the transformation frequency qualified. Independent transformed colonies are obtained at a frequency of 1 in 104 to 1 in 105 cells when integrating plasmids are introduced using calcium phosphate coprecipitation. The frequency is about tenfold higher when extrachromosomal shuttle vectors are introduced into cells.
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  • 44
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 410-417 
    ISSN: 0192-253X
    Keywords: Suppressor †RNA genes ; yeast ; lacZ expression in D. discoideum ; †RNATrp genes ; †RNAGlu genes ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We describe the generation of Dictyostelium discoideum cell lines that carry different suppressor †RNA genes. These genes were constructed by primer-directed mutagenesis changing a †RNATrp(CCA) gene from D. discoideum to a †RNATrp(amber) gene and changing a †RNAGlu(UUC) gene from D. discoideum to a †RNAGlu(ochre) as well as a †RNAGlu(amber) gene. These genes were stably integrated into the D. discoideum genome together with a reporter gene. An actin 6::lacZ gene fusion carrying corresponding translational stop signals served as a reported. Active β-galactosidase is expressed only in D. discoideum strains that contain, in addition to the reporter, a functional suppressor †RNA. Both amber suppressors are active in D. discoideum without interfering significantly with cell growth and development. We failed, however, to establish cell lines containing a functional †RNAGlu(ochre) suppressor. This may be due to the fact that nearly every message from D. discoideum known so far terminates with UAA. Therefore a †RNA capable of reading this termination codon may not be compatible with cell growth.
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  • 45
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 418-424 
    ISSN: 0192-253X
    Keywords: Transcription ; development ; cAMP ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Genetic studies have revealed a network of three unlinked regulatory genes that control the developmental expression of the family of endogenous lectins in Dictyostelium discoideum. Mutations in the disA and disB loci have a null phenotype and do not express the discoidin I or II lectins. The third mutation, drsA, is a second-site suppressor of the disB mutation, which restores expression of all lectin species. Cells carrying this mutation express the discoidin lectins during growth, which is in contrast to wild-type cells in which lectin synthesis is developmentally regulated. In addition to this basic level of genetic control, the conditions of growth dramatically influence the patterns of discoidin expression. Growth-phase wild-type cells do not express lectin if the cells are grown on plates in association with bacteria. However, wild-type cells growing in bacterial suspensions express high levels of lectin during growth. Synthesis of the discoidin lectins in growing cells is sensitive to the levels of extracellular cyclic adenosine monophosphate (cAMP) and folic acid. These results suggest that the drsA mutation renders cells insensitive to cAMP and/or folate and thus allows expression of lectin during growth.
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  • 46
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 439-441 
    ISSN: 0192-253X
    Keywords: Cellular slime molds ; timing of differentiation ; differentiation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Vital dyes have been extensively used in many laboratories to distinguish the prestalk and prespore zones of a migrating slug in Dictyostelium discoideum. Here we present evidence that the two zones do not form immediately following aggregation, as is sometimes assumed, but only after 1-4.5 hr of migration. When the transition to the two-toned state occurs, it changes very quickly, in a matter of approximately 10 min. Furthermore, it can be rapidly induced by submerging the slug in mineral oil. The time of appearance of the zones is correlated with previously reported changes in the distribution of cyclic AMP secretion and with changes in regulation properties along the axis of the slug.
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  • 47
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 442-446 
    ISSN: 0192-253X
    Keywords: Prestalk cells ; prespore cells ; slugs ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Ammonia appears to be an important regulatory signal for several aspects of the Dictyostelium life cycle. The postulated role of ammonia in the determination of the prespore pathway in cells of the slug stage has led us to examine the effect of ammonia on the prestalk/prespore ratio of migrating slugs. In the presence of 10-3 M ammonium chloride, the volume of the prestalk region decreases by 40.8%. The kinetics of the process make it unlikely that this is due to a shift in the differentiation pathway. A test of the hypothesis that the decrease in volume of the prestalk region is due to the conversion of prestalk cells to anterior-like cells shows that the percent of anterior-like cells in the posterior region increases by the amount predicted by the hypothesis. This suggests that ammonia may be the molecular signal, produced by the tip, that prevents anterior-like cells from chemotactically migrating to the tip and thereby becoming anterior cells. The effect of enzymatic removal of ammonia from vitally stained migrating slugs is the appearance of a series of dark stripes beginning at the posterior end and progressing forward. We interpret this as a result of progressive removal of anterior-like cells from tip dominance and essentially as the formation of new potential tips. Indeed, in a few cases one or even two of the stripes separate from the posterior of the cell mass and form small fruiting bodies. We consider the phenomenon of stripe formation further evidence that the tip acts on anterior-like cells through ammonia.
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  • 48
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 427-438 
    ISSN: 0192-253X
    Keywords: Cellular slime molds ; patterning ; development ; immunoblotting ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A series of monoclonal antibodies were previously raised against developing Polysphondylium pallidum cells. In this work, six of these antibodies have been used as probes to identify and characterize antigens regulated during development. Soluble and membrane fractions of P. pallidum cells at six stages of development or three stages of cyclic adenosine monophosphate (cAMP)-induced development were run in sodium dodecyl sulfate (SDS)-polyacrylamide gels and subjected to Western blot analysis. Three of the monoclonals, anti-Tp200, anti-Tp423, and antiPg 101, stain sorogen tips. Tp423 and Tp200 are membrane-associated antigens; both are stable to urea extraction, and Tp200 remains in the membrane after NaOH extraction. Tp423 is present in starved cells but is more prominent in sorogens and particularly in cAMP-developed cells. In contrast, Tp200 is first detected in early to mid-aggregation and is more abundant late in development. Pg101, which is expressed as a gradient with its highest concentration in tips, first appears in tight aggregates but is much more abundant in sorogens; unlike the Tp antigens, Pg101 is not greatly induced in cAMP-developed cells. All three of these antigens undergo changes in apparent molecular weight at the tight aggregate or sorogen stage: The gel mobilities of Tp200 and Pg101 increase, whereas that of Tp423 decreases. In addition to the tip-specific monoclonals, two monoclonals that stain all but the tips of sorogens have been used for analysis. One of these, anti-3D 10Pnk stains most cells within secondary tips, whereas anti-3D 10Dif does not. 3D 10Dif is membrane associated; it is present very early in development, increasing two- to threefold through the sorogen stage and diminishing in late cAMP-developed cells. 3D 10Pnk is a mostly soluble species first detected in late streaming. Anti-1c3, a sixth monoclonal, which stains nuclei uniformly throughout sorogens, is also developmentally expressed. 1c3 is mainly membrane associated and is expressed from late streaming through the sorogen stage.
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  • 49
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 453-453 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 50
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 447-452 
    ISSN: 0192-253X
    Keywords: Pattern regulation ; cell sorting ; transplantation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In classical transplantation experiments, we have attempted to distinguish between cell sorting and position-dependent mechanisms of pattern formation during the regulation of amputated prespore regions of Dictyostelium slugs. Host and transplanted tissue were distinguished by prior labelling with 14C and 3H, and their distributions in the manipulated slugs were determined by double-label scintillation counting of serial sections of the slugs. First, we showed that the regenerated prestalk region is formed predominantly from the anterior of a prespore isolate. This disproves extreme cell sorting models, which hold that the new prestalk cells arise at random in the prespore mass, before sorting to the prestalk zone. Second, we showed that a proportion of posterior cells, which are not normally recruited into the new prestalk region, can be recruited into it if they are transplanted into the anterior of the prespore isolate. Recruitment of these cells occurs only during the first hour of regulation. We believe that these experiments prove a position-dependent mechanism of pattern formation during slug regulation. This in turn implies the existence of localized inductive signals in the regulating fragments. Although cell sorting occurs efficiently when cells are misplaced in the slug, it does not appear to play a major role in reforming the prestalk/prespore pattern.
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  • 51
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    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Computational Chemistry 11 (1990) 
    ISSN: 0192-8651
    Keywords: Computational Chemistry and Molecular Modeling ; Biochemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Computer Science
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  • 52
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    Journal of Computational Chemistry 11 (1990), S. 1-18 
    ISSN: 0192-8651
    Keywords: Computational Chemistry and Molecular Modeling ; Biochemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Computer Science
    Notes: Counterpoise corrected ab initio calculations are reported for (H2O)2 and H2O-H2CO. Geometry searches were done in the moment-optimized basis DZP' at the SCF, MP2, and CEPA-1 levels of theory, followed by more accurate single-point calculations in basis ESPB, which includes bondfunctions to saturate the dispersion energy. The final equilibrium binding energies obtained are -4.7 ±0.3 kcal/mol for a near-linear (H2O)2 structure and -4.6 ±0.3 kcal/mol for a strongly bent HOH ‥ OCH2 structure. The energy difference between these systems is much smaller than in all previous ab initio work. Cyclic (C2h) and bifurcated (C2v) transition structures for (H2O)2 are located at 1.0 ±0.1 kcal/mol and 1.9 ±0.3 kcal/mol above the global minimum, respectively. A new partitioning scheme is presented that rigorously partitions the MP2 correlation interaction energy in intra and intermolecular (dispersion) contributions. These terms are large (up to 2 kcal/mol) but of opposite sign for most geometries studied and hence their overall effect upon the final structures is relatively small. The relative merits of the MP2 and CEPA-1 approaches are discussed are discussed and it is concluded that for economical reasons MP2 is to be preferred, especially for larger systems.
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  • 53
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    Journal of Computational Chemistry 11 (1990), S. 45-57 
    ISSN: 0192-8651
    Keywords: Computational Chemistry and Molecular Modeling ; Biochemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Computer Science
    Notes: A data compression method is presented that is generally applicable to first-order convergent iterative procedures that employ subspace expansions or extrapolations based on successive correction vectors. This method is based on the truncation of insignificant information in successive correction vectors. Although the correction vectors themselves may be severely truncated with the proposed approach, the final solution vector may be represented to arbitrary accuracy. A feature of the proposed method is that more slowly convergent iterative procedures allow the correction vectors to be more severely truncated without affecting the overall convergence rate. The method is implemented and applied to the iterative Davidson diagonalization method. If the compressed representation of the expansion vectors can be held in main computer memory, then a significant reduction in the I/O requirements is achieved.
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  • 54
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    Journal of Computational Chemistry 11 (1990), S. 67-75 
    ISSN: 0192-8651
    Keywords: Computational Chemistry and Molecular Modeling ; Biochemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Computer Science
    Notes: By examining the displacement coordinate metric three modes of constrained optimization for large molecules and clusters are suggested. The first method corresponds to a conventional optimization using internal coordinates. The second mode has applications with respect to both internal and cartesian coordinates. The final mode is particularly interesting because it can result in computational savings. A mixture of both internal and cartesian coordinates is specified where these coordinates are usually a subset of the molecules or clusters total coordinate set. In the optimization only a subset of the energy derivatives need be evaluated reducing the computational effort associated with the gradient calculation.
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  • 55
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    Journal of Computational Chemistry 11 (1990), S. 82-87 
    ISSN: 0192-8651
    Keywords: Computational Chemistry and Molecular Modeling ; Biochemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Computer Science
    Notes: A finite-field method for the calculation of polarizabilities and hyperpolarizabilities is developed based on both an energy expansion and a dipole moment expansion. This procedure is implemented in the MOPAC semiempirical program. Values and components of the dipole moment (μ), polarizability (α), first hyperpolarizability (β), and second hyperpolarizability (γ) are calculated as an extension of the usual MOPAC run. Applications to benzene and substituted benzenes are shown as test cases utilizing both MNDO and AM1 Hamiltonians.
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  • 56
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    Journal of Computational Chemistry 11 (1990), S. 121-151 
    ISSN: 0192-8651
    Keywords: Computational Chemistry and Molecular Modeling ; Biochemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Computer Science
    Notes: A procedure has been developed for global energy minimization of surface loops of proteins in the presence of a fixed core. The ECEPP potential function has been modified to allow more accurate representations of hydrogen bond interactions and intrinsic torsional energies. A computationally efficient representation of hydration free energy has been introduced. A local minimization procedure has been developed that uses a cutoff distance, minimization with respect to subsets of degrees of freedom, analytical second derivatives, and distance constraints between rigid segments to achieve efficiency in applications to surface loops. Efficient procedures have been developed for deforming segments of the initial backbone structure and for removing overlaps. Global energy minimization of a surface loop is accomplished by generating a sequence (or a trajectory) of local minima, the component steps of which are generated by searching collections of local minima obtained by deforming seven-residue segments of the surface loop. The search at each component step consists of the following calculations: (1) A large collection of backbone structures is generated by deforming a seven-residue segment of the initial backbone structure. (2) A collection of low-energy backbone structures is generated by applying local energy minimization to the resulting collection of backbone structures (interactions involving side chains that will be searched in this component step are not included in the energy). (3) One low-energy side-chain structure is generated for each of the resulting low-energy backbone structures. (4) A collection of low-energy local minima is generated by applying local energy minimization to the resulting collection of structures. (5) The local minimum with the lowest energy is retained as the next point of the trajectory. Applications of our global search procedure to surface segments of bovine pancreatic trypsin inhibitor (BPTI) and bovine trypsin suggest that component-step searches are reasonably complete. The computational efficiency of component-step searches is such that trajectories consisting of about 10 component steps are feasible using an FPS-5200 array processor. Our procedure for global energy minimization of surface loops is being used to identify and correct problems with the potential function and to calculate protein structure using a combination of sequence homology and global energy minimization.
    Additional Material: 7 Ill.
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  • 57
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    Journal of Computational Chemistry 11 (1990), S. 416-430 
    ISSN: 0192-8651
    Keywords: Computational Chemistry and Molecular Modeling ; Biochemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Computer Science
    Notes: A systematic analysis of the molecular electrostatic potential (MEP) is presented. This study has been performed with a twofold purpose: first, to study the MEP dependence with regard to the quality of the basis set used to compute the ab initio SCF wavefunction and second, to develop and to assess a new strategy for computing isoelectrostatic potential maps using the semiempirical MNDO wavefunction. The only differences between this procedure and the ab initio SCF MEP computation lie in the freezing of the inner electrons and in the origin of the first-order density matrix. The statistical analysis of MEPs computed for a large number of molecules from MNDO wavefunction and ab initio SCF wavefunctions obtained using STO-3G, 4-31G, 6-31G, 4-31G*, 6-31G*, and 6-31G** basis sets points out the ability of any wavefunction to reproduce the general topological characteristics of the MEP surfaces. Nevertheless, split-valence basis sets including polarization functions are necessary to obtain accurate MEP minimum energy values. MNDO wavefunction tends to overestimate the MEP minima depth by a constant factor and shows an excellent ability to reflect the relative variation of MEP minima energies derived from a rather sophisticated (6-31G*) basis set, lacking of the shortcomings detected in the semiempirical CNDO approximation.
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  • 58
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    Journal of Computational Chemistry 11 (1990), S. 487-492 
    ISSN: 0192-8651
    Keywords: Computational Chemistry and Molecular Modeling ; Biochemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Computer Science
    Notes: Expressions are derived for computing the kinetic energy of a peptide with fixed geometry, in terms of dihedral angles as generalized coordinates; other expressions required for the solution of Lagrange's equations are also presented. The peptide is regarded as held stationary at one end. We also outline the computations that are needed in calculating the components of the third derivative of a potential energy function that consists of a sum of pairwise interatomic interactions.
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  • 59
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    Journal of Computational Chemistry 11 (1990), S. 819-828 
    ISSN: 0192-8651
    Keywords: Computational Chemistry and Molecular Modeling ; Biochemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Computer Science
    Notes: Complexes of alkali and alkaline earth cations with organic compounds are modeled by describing ionligand interaction energies with pair potentials and intraligand as well as interligand energies with the MM2 potential. New pair potentials for the interaction of Li+, Na+, K+, Mg2+, and Ca2+ ions are derived on the basis of 30,000 ab initio interaction energy values with 70 selected model ligand molecules. Various problems of the combination of these two basically different potentials are discussed. An application for the K+ complex of 18-crown-6 is presented. For more flexible ligands the introduction of three-body correction terms of the pair potentials seems necessary.
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  • 60
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    Journal of Computational Chemistry 11 (1990), S. 924-929 
    ISSN: 0192-8651
    Keywords: Computational Chemistry and Molecular Modeling ; Biochemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Computer Science
    Notes: The geometry, electronic structure, and photoelectron spectra of Sb2 and Sb4 have been investigated employing the LCGTO-MP-LSD method. For both molecules the geometrical and spectroscopic constants computed using the VWN potential are in very good agreement with the available experimental data. The HeI photoelectron spectra were computed with different potentials (Xα, GL, HL, BH, and VWN). The calculated spectra are in excellent agreement with the experimental ones and the use of the VWN potential gives the most accurate results. The reliable results for molecules complement those for the Sb atom and show that the proposed LSD Model Potential is able to give an accurate description of the structural and spectroscopic properties of the title molecules.
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  • 61
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    Journal of Computational Chemistry 11 (1990), S. 958-971 
    ISSN: 0192-8651
    Keywords: Computational Chemistry and Molecular Modeling ; Biochemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Computer Science
    Notes: Intermolecular potential functions have been developed for use in computer simulations of alkyl ethers. The simple OPLS model was adopted and parameterized to yield good descriptions of bimolecular and ion-molecule complexes as well as to reproduce experimental thermodynamic properties of liquid ethers. The principal testing featured Monte Carlo statistical mechanics simulations for liquid dimethyl ether (DME), ethyl methyl ether (EME), diethyl ether (DEE), and tetrahydrofuran (THF). Average errors of 1-3% are obtained for the computed densities and heats of vaporization including results for THF at pressures up to 5000 atm. The torsional motion about the central C—O bonds in EME and DEE was included in the simulations using rotational potential functions fit to results of molecular mechanics (MM2) calculations. The liquid-state environment is found to have negligible effect on the conformational equilibria.
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  • 62
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    Journal of Computational Chemistry 11 (1990), S. 972-977 
    ISSN: 0192-8651
    Keywords: Computational Chemistry and Molecular Modeling ; Biochemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Computer Science
    Notes: A quadrature-point-driven implementation of the standard Rys polynomial method for computing two-electron repulsion integrals of gaussian basis functions has been found to be both concise and openended with respect to the angular momentum of the gaussian functions (i.e., s,p,d,f,g,…). These are important features in certain applications, such as molecular properties and property gradients.
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  • 63
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    Journal of Computational Chemistry 11 (1990), S. 994-1002 
    ISSN: 0192-8651
    Keywords: Computational Chemistry and Molecular Modeling ; Biochemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Computer Science
    Notes: Free energy perturbation calculations were performed to determine the free energy of binding associated with the presence of perhaps an unusual hydroxyl group in the transition state analog of nebularine, an inhibitor of the enzyme adenosine deaminase. The presence of a single hydroxyl group in this inhibitor has been found to contribute -9.8 kcal/mol to the free energy of binding, with a 108-fold increase in the binding affinity by the enzyme. In this work, we calculate the difference in solvation free energy for the 1,6-dihydropurine complex versus that of the 6-hydroxyl-1,6-dihydropurine complex to determine if this marked increase in binding affinity is attributed to an unusually hydrophobic hydroxyl group. The calculated ΔG associated for the solvation free energy is -11.8 kcal/mol. This large change in the solvation free energy suggests that this hydroxyl is instead unusually hydrophilic and that the difference in free energy of interaction for the two inhibitors to the enzyme must be at least ca. 20 kcal/mol. Although the crystal structure for adenosine deaminase is currently not known, we attempt to mimic the nature of the active site by constructing models which simulate the enzyme-inhibitor complex. We present a first attempt at determining the change in free energy of binding for a system in which structural data for the enzyme is incomplete. To do this, we construct what we believe is a minimal model of the binding between adenosine deaminase and an inhibitor. The active site is simulated as a single charged carboxyl group which can form a hydrogen bond with the hydroxyl group of the analog. Two different carboxyl anion models are used. In the first model, the association is modeled between an acetic acid anion and the modified inhibitor. The second model consists of a hydrophobic amino acid pocket with an interior Glu residue in the active site. From these models we calculate the change in free energy of association and the overall change in free energy of binding. We calculate the free energies of interaction both in the absence and presence of water. We conclude from this that the presence of a single suitably placed-CO-2 group probably cannot explain the binding effect of the-OH group and that additional interactions will be found in the adenosine deaminase active site.
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  • 64
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    Journal of Computational Chemistry 11 (1990), S. 1029-1037 
    ISSN: 0192-8651
    Keywords: Computational Chemistry and Molecular Modeling ; Biochemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Computer Science
    Notes: Pseudopotentials and valence basis sets to be used in calculations for organometallic compounds of zinc and magnesium have been tested in calculations for the M(CH3)n (M = Zn, Mg; n = 1,2) molecules. Valence correlation effects are treated at the SDCI and CEPA levels. The capability of a polarization potential on zinc to account for the valence shell contracting effect of core valence correlation is studied. Properties considered are geometries, force constants, Mulliken populations, ionization potentials, atomization, and binding energies. Differences in bonding between the two dimethyl compounds are discussed.
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  • 65
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    Journal of Computational Chemistry 11 (1990), S. 1094-1100 
    ISSN: 0192-8651
    Keywords: Computational Chemistry and Molecular Modeling ; Biochemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Computer Science
    Notes: The acid-catalyzed hydrolysis mechanisms of maleamic (4-amino-4-oxo-2-butenoic) acids were studied using AM1 method. The reaction proceeded mainly in two steps: (1) nucleophilic attack of the undissociated carboxyl group on the adjacent aminocarbonyl carbon via a zwitterionic intermediate; and (2) the rate-determining proton transfer to form the zwitterionic tetrahedral intermediate. In each step, the hydration of water and hydronium ion molecules was important in stabilizing the polarized intermediates. The substituent effects at the amide moiety and the 2,3-positions of the maleamic acids were qualitatively estimated for each step.
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  • 66
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    Journal of Computational Chemistry 11 (1990) 
    ISSN: 0192-8651
    Keywords: Computational Chemistry and Molecular Modeling ; Biochemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Computer Science
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  • 67
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    Journal of Computational Chemistry 11 (1990), S. 1139-1146 
    ISSN: 0192-8651
    Keywords: Computational Chemistry and Molecular Modeling ; Biochemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Computer Science
    Notes: A prescription for applying the method of molecular similarity calculations based on electrostatic potentials and fields is developed by consideration of a typical structure-activity series. Firm conclusions are drawn about the nature of the grid of points surrounding the molecules and about the choice of geometry, but options for point charges are less clearcut.
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  • 68
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    Journal of Computational Chemistry 11 (1990), S. 899-908 
    ISSN: 0192-8651
    Keywords: Computational Chemistry and Molecular Modeling ; Biochemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Computer Science
    Notes: An expanded form of the Wiener number is suggested for characterization of molecular graphs and structure-property correlations. The simple, computer-oriented method for counting of the novel index is briefly discussed.
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  • 69
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    Journal of Computational Chemistry 11 (1990), S. 1193-1205 
    ISSN: 0192-8651
    Keywords: Computational Chemistry and Molecular Modeling ; Biochemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Computer Science
    Notes: A common requirement in conformational analysis is the identification of a molecule's lowest energy conformations. The application of the A* algorithm to this problem is examined. The algorithm uses heuristic information about the problem domain to direct the search and has been implemented in a system for performing automated conformational analysis. The method is detailed and sample results presented. Some limitations of the approach are identified.
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  • 70
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    Journal of Computational Chemistry 11 (1990), S. 1009-1016 
    ISSN: 0192-8651
    Keywords: Computational Chemistry and Molecular Modeling ; Biochemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Computer Science
    Notes: Heats of formation and net atomic charges of some 120 structures involving substituted phenols, anilines, and benzoic acids and the corresponding anions were calculated by MNDO, AM1, and PM3 semiempirical methods. The gas phase acidities of substituted phenols and anilines and the net atomic charges on the anionic heteroatoms of the corresponding anions have been successfully correlated with σ- constants. Moreover, good correlations with σ were found for the charges on the acidic hydrogens of substituted phenols and anilines. In contrast, the gas phase acidities of substituted benzoic acids and the charges on the anionic oxygens of the corresponding anions are better correlated with Taft σ° constants. Comparisons of these results with experimental data and ab initio theoretical calculations indicate that AM1 and PM3 methods are much better than MNDO in predicting the acidity of aromatic compounds.
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  • 71
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    Journal of Computational Chemistry 11 (1990), S. 1038-1046 
    ISSN: 0192-8651
    Keywords: Computational Chemistry and Molecular Modeling ; Biochemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Computer Science
    Notes: We studied geometries and energies of complexes between water and neutral or protonated imidazole by ab initio molecular orbital calculations using the 4-31G basis set with and without the counterpoise correction. Positions of hydration sites and relative binding energies could be also estimated by using the electrostatic field map of imidazole as calculated by our bond increment method. The reliability of the calculations is confirmed by comparing the geometries of the imidazole-water complex to the experimental ones from the Cambridge Structural Database. These were obtained by X-ray diffraction studies on crystals with water bound to a molecule containing the imidazole fragment.
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  • 72
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    Journal of Computational Chemistry 11 (1990), S. 1061-1071 
    ISSN: 0192-8651
    Keywords: Computational Chemistry and Molecular Modeling ; Biochemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Computer Science
    Notes: A new method for the evaluation of metal complexes with molecular mechanics calculations is described. The method has been employed to determine the global minimum energy conformations of three seven-coordinate lanthanide complexes. The method searches the potential energy surface of the metal complexes for the global minimum structure using the recently reported Random Incremental Pulse Search (RIPS). The molecular mechanics calculations were performed using the MM2 metal-extended force field (MM2MX). This force field excludes 1-3 bending terms about the metal center and, instead, explicitly includes 1-3 nonbonded van der Waals interactions. This affords a model based mainly on steric interactions about the ligating atoms, and removes the necessity of specifying ligand-metal-ligand bond angles for seven-coordinate and higher metal complexes in which no preferred bond angle can be proficiently determined. The calculated minimum-energy structures were well known seven-coordinate conformations in all cases, and a comparison with X-ray crystallographic is presented.
    Additional Material: 9 Ill.
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  • 73
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    Journal of Computational Chemistry 11 (1990), S. 1076-1079 
    ISSN: 0192-8651
    Keywords: Computational Chemistry and Molecular Modeling ; Biochemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Computer Science
    Notes: The method of approximation of the frozen molecular fragment (FMF) we derived has been applied to calculations of proton affinities. Results are in good agreement with experimental data and extended basis set calculations.
    Additional Material: 3 Tab.
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  • 74
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    Journal of Computational Chemistry 11 (1990), S. 1087-1093 
    ISSN: 0192-8651
    Keywords: Computational Chemistry and Molecular Modeling ; Biochemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Computer Science
    Notes: Myelin basic protein (MBP) is the major extrinsic protein of the myelin sheath in the central nervous system. It is this protein that is destroyed in such demyelinating diseases as multiple sclerosis. We have examined the predicted structures of one segment of MBP using the molecular mechanics program ECEPP83 developed by Scheraga and co-workers as modified by Chuman, Momany, and Schafer. We have focused upon a segment, 87-118, containing the Pro-Pro-Pro sequence (residues 100-102), which has been predicted from standard algorithms to exist in a hairpin loop connecting anti-parallel beta-strands. Several local energy minima have been found and reported. Tripoline sequences are not rare in proteins, but their structure and function is still uncertain.
    Additional Material: 4 Ill.
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  • 75
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    Journal of Computational Chemistry 11 (1990), S. 1101-1110 
    ISSN: 0192-8651
    Keywords: Computational Chemistry and Molecular Modeling ; Biochemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Computer Science
    Notes: Two new computational methods for the evaluation and partitioning of projected, planar, and averaged slab electron densities, implemented in the program PRODEN, are presented. The new algorithms for the projection, demarcation, and integration of electron densities are described and evaluated in terms of speed and numerical accuracy. Integrated Projected Populations are analyzed and some of the advantages and limitations of the methods are discussed.
    Additional Material: 4 Ill.
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  • 76
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    Journal of Computational Chemistry 11 (1990), S. 930-942 
    ISSN: 0192-8651
    Keywords: Computational Chemistry and Molecular Modeling ; Biochemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Computer Science
    Notes: The effect of the addition of diffuse functions of sp type on the first row atoms (and/or of d type on phosphorus) to a MINI-1 minimal basis set is evaluated by comparing the SCF description of the interaction energy and its decomposition, counterpoise (CP) corrected and uncorrected for basis set superposition errors (BSSEs), with that produced by extended basis sets (6-31G** and 3-21G+) as well as by basis sets (6-31G** + VPS and (2d)S), previously successfully tested, designed to minimize the BSSE. The systems considered are a few anions (F-, HCOO-, H2PO-4) H-bonded to water; the basis set performance on a different geometry (bifurcated instead of linear) of the phosphate-water adduct was also considered. An additional comparison with the interaction energy in a neutral dimer, (HF)2, was carried out. The addition of diffuse functions to the MINI-1 basis set, instead of resorting to extended basis sets, seems to be a sensible choice for anions. The equilibrium distance is shifted towards the reference basis set value at the CP corrected level as well as at the uncorrected one. The spread of the energy due to the basis set is hardly distinguishable when the CP corrected values are plotted. The description of the interaction energy in dimers involving phosphorus is improved using d type diffuse functions.
    Additional Material: 11 Ill.
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  • 77
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    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Computational Chemistry 11 (1990) 
    ISSN: 0192-8651
    Keywords: Computational Chemistry and Molecular Modeling ; Biochemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Computer Science
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  • 78
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    Journal of Computational Chemistry 11 (1990), S. 1017-1020 
    ISSN: 0192-8651
    Keywords: Computational Chemistry and Molecular Modeling ; Biochemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Computer Science
    Notes: Ab initio CI calculations using pseudopotentials to describe germanium inner electrons are carried out on the low-lying excited singlet states (T2) of GeH4. A theoretical analysis of these states in terms of Mulliken population of Rydberg orbitals for each state and oscillator strengths allow us to reinterpret its experimental Vacuum UV electronic spectrum.
    Additional Material: 3 Tab.
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  • 79
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    Journal of Computational Chemistry 11 (1990), S. 1047-1060 
    ISSN: 0192-8651
    Keywords: Computational Chemistry and Molecular Modeling ; Biochemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Computer Science
    Notes: The algorithm used by the program GEPOL to compute the Molecular Surface (MS), as defined by Richards, is presented in detail. GEPOL starts like other algorithms from a set of spheres with van der Waals radii, centered on the atoms or group of atoms of the molecule. GEPOL computes the MS by first searching the spaces inaccessible to the solvent and consequently filling them with a new set of spheres. Here we study the behavior of the method with its parameters, presenting several examples of application.
    Additional Material: 13 Ill.
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  • 80
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    Journal of Computational Chemistry 11 (1990), S. 1080-1086 
    ISSN: 0192-8651
    Keywords: Computational Chemistry and Molecular Modeling ; Biochemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Computer Science
    Notes: A boundary element method is developed to compute the electrostatic potential inside and around molecules in an electrolyte solution. A set of boundary integral equations are derived based on the integral formulations of the Poisson equation and the linearized Poisson-Boltzmann equation. The boundary integral equations are then solved numerically after discretizing the molecular surface into a number of flat triangular elements. The method is applied to a spherical molecule for which analytical solutions are available. Use is made of both constant and linearly varying unknowns over the boundary elements, and the method is tested for various values of parameters such as the dielectric constant of the molecule, ionic strength, and the location of the interior point charge. The use of the boundary integral method incorporating the nonlinear Poisson-Boltzmann equation is also briefly discussed.
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  • 81
    ISSN: 0749-503X
    Keywords: Hansenula polymorpha ; methanol ; glycerol ; dihydroxyacetone ; xylose ; alcohol oxidase ; continuous culture ; regulation ; mutants ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The physiological responses of Hansenula polymorpha wild-type and mutant strains 17B (dihydroxyacetone kinase-negative) and 17BG51 (dihydroxyacetone kinase- and glycerol kinase-negative) to growth on mixtures of xylose and methanol in chemostats were investigated. Increasing methanol concentrations (0-110 mM) in the feed of the wild-type culture resulted in increasing cell densities and a gradual switch towards methanol metabolism. At the lower methanol feed concentrations the mutant cultures used methanol and xylose to completion and changes in enzyme patterns comparable to the wild type were observed. This was not reflected in significant changes in cell densities. Instead, formaldehyde assimilation resulted in dihydroxyacetone (DHA) production, which was proportional to the amount of methanol added. At intermediate methanol concentration the cultures showed a strong variation in DHA levels and cell densities. Further increased in the methanol feed concentrations resulted in a drop in DHA accumulation rates, repression of alcohol oxidase synthesis and accumulation of residual methanol. The phenomena were studied in more detail in transition experiments and with gradients of methanol. The results indicate that xylulose-5-phosphate (Xu5P) generated in xylose metabolism served as acceptor molecule for formaldehyde assimilation by the peroxisomal enzyme DHA synthase. Accumulation of DHA in the mutant cultures, however, further diminished the availability of carbon for growth. The data suggest that with increasing methanol concentrations Xu5P eventually became growth rate limiting. This resulted in an unstable situation but wash-out of the culture did not occur to a significant extent. Instead, DHA accumulation ceased and cell densities, and enzymes specifically involved in xylose metabolism increase, indicating that the organism resumed its xylose metabolism. The molecular mechanisms controlling the partitioning of Xu5P over xylose (pentose phosphate pathway) and methanol (peroxisome) metabolism under these conditions remain to be elucidated.
    Additional Material: 4 Ill.
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  • 82
    ISSN: 0749-503X
    Keywords: Hansenula polymorpha ; methanol ; dihydroxyacetone ; xylose ; mutants ; transketolase ; formaldehyde ; continuous culture ; peroxisome ; regulation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Contrary to expectation, a mutant of Hansenula polymorpha blocked in dihydroxyacetone (DHA) synthase was able to assimilate methanol-carbon when grown in chemostat culture on mixtures of xylose and methanol. Incubation of a DHA synthase- and DHA kinase-negative double mutant resulted in DHA accumulation, indicating that a DHA synthase-type of reaction was involved. Low residual DHA synthase activity subsequently was shown to be present when using an assay with improved sensitivity. This activity was not associated with the (mutated) DHA synthase protein, which was still present in the peroxisomes, but with the enzyme transketolase. Transketolase from methanol grown cells was purified (525-fold) to homogeneity in 9% yield. The native enzyme was dimeric, as has been reported fro other transketolases, with a subunit molecular weight of 74000. The affinity of the purified enzyme for formaldehyde was low (Km = 5 mM), but high for xylulose-5-phosphate (ca. 10 μM). The in vivo functioning of transketolase in formaldehyde assimilation, and the influence of the hydration state of formaldehyde is discussed.
    Additional Material: 4 Ill.
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  • 83
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 84
    ISSN: 0749-503X
    Keywords: linear plasmids ; killer ; stability ; mating type locus control ; a1-α2 repressor ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The linear killer plasmids, pGKL1 and pGKL2, from Kluyveromyces lactis stably replicated in mitochondria1 DNA-deficient (ρ0) MATa or MATα haploids of Saccharomyces cerevisiae, but were unstable and frequently lost in ρ° MATa/MATα diploids, suggesting that the replication of pGKL plasmids was under the control of the MAT locus. In MATa/MATα cells of S. cerevisiae, the MATα gene product (α2) is combined with the MATa gene product (a1) and the resultant protein, a1-α2, acts to repress the expression of haploid specific genes. Experiments showed that the K. lactis linear plasmids were stably maintained in ρ0 mata1/MATα diploids, indicating that the a1-α2 repressor interfered with the stability of pGKL2. It was revealed by computer analysis that the consensus sequence homologous to the a1-α2 repressor binding site occurred within the coding regions of pGKL2 genes which were presumed to be essential for the plasmid replication. Since the plasmids were stably maintained in diploids of K. lactis, the mating type control must not be working there.
    Additional Material: 2 Ill.
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  • 85
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    Yeast 6 (1990), S. 455-460 
    ISSN: 0749-503X
    Keywords: Ornithine decarboxylase ; putrescine ; SPE1 gene ; spe10 mutation ; chromosome XI ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The gene for ornithine decarboxylase in Saccharomyces cerevisiae, SPE1, has been assigned to chromosome XI by the technique of transverse alternating pulsed field electrophoresis and DNA-DNA hybridization. Genetic mapping by tetrad analysis shows that the SPE1 gene is located on the left arm of chromosome XI, 6 cM from the LAP1 gene and 43 cM from the TRP3 gene. The spe10 mutation previously isolated in this laboratory is mapped to the N-terminal region of the SPE1 gene, and therefore should be designated as a spe1 allele.
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  • 86
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    Yeast 6 (1990) 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 87
    ISSN: 0749-503X
    Keywords: Omnipotent suppressor ; gene structure ; evolutionary conservation ; codon bias analysis ; Pichia pinus ; yeast ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: SUP2(SUP35) is an omnipotent suppressor gene, coding for an EF-1α-like protein factor, intimately involved in the control of translational accuracy in yeast Saccharomyces cerevisiae.In the present study a SUP2 gene analogue from yeast Pichia pinus was isolated by complementation of the temperature-sensitive sup2 mutation of S. cerevisiae.The nucleotide sequence of the SUP2 gene of P. pinus codes for a protein of 82·4 kDa, exceeding the Sup2 protein of S. cerevisiae by 6 kDa. Like the SUP2 gene product of S. cerevisiae, the Sup2 protein of P. pinus represents a fusion of a unique N-terminal part of a region homologous to EF-1α. The comparison of amino acid sequences of the Sup2 proteins reveals high conservations (76%) of the C-terminal region and low conservation (36%) of the N-terminal part where, in addition, the homologous correspondence is ambiguous.Proteins related to the Sup2 of S. cerevisiae where found in P. pinus and some other yeast species by the immunoblotting technique.The relation between the evolutionary conservation of different regions of the Sup2 protein and their functional significance is discussed.
    Additional Material: 7 Ill.
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  • 88
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    Yeast 6 (1990), S. 535-536 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 89
    ISSN: 0749-503X
    Keywords: Cell wall porosity ; permeability ; mannan ; cell wall composition ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The cell porosity of batch-grown Saccharomyces cerevisiae was maximal in the early exponential phase and fell off rapidly to lower levels in later growth phases.Treatment of stationary-phase cells with alpha-mannosidase restored wall porosity to the level of cells in early exponential phase. When cells in the early exponential phase were treated with alpha-mannosidase, or tunicamycin, an inhibitor of N-glycosylation, even higher porosities were obtained. Mutants with truncated mannan side-chains in their wall proteins also had very porous walls. The importance of the mannan side-chains for wall porosity was also seen during sexual induction. Treatment with alpha pheromone, which leads to the formation of wall proteins with shorter mannan side-chains, enhanced wall porosity.Disulphide bridges also affect cell wall porosity. They were predominantly found in the glucanase-soluble wall proteins. Because the main part of the mannan side-chains is also found in this family of wall proteins, our results demonstrate that the glucanase-soluble mannoproteins limit cell wall porosity in yeast.
    Additional Material: 1 Ill.
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  • 90
    ISSN: 0749-503X
    Keywords: Hansenula polymorpha ; peroxisomes ; peroxisome-deficient mutants ; amine oxidase ; D-amino acid oxidase ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have studied growth of two peroxisome-deficient mutant strains of Hansenula polymorpha on glucose in the presence of different organic nitrogen sources (methylamine, ethylamine and D-alanine), the metabolism of which is mediated by peroxisome-borne oxidases in wild-type (WT) cells. Both strains grew well on each of these substrates with growth rates comparable to WT cells. Growth on both methylamine and ethylamine was associated with enhanced levels of catalase and amine oxidase in the cells; in D-alanine-grown cells D-amino acid oxidase activity and increased. In WT cells of H-polymorpha the activities of these enzymes were confined to the peroxisomal matrix; however, in both peroxisome-deficient strains their activities were localized in the cytosol. Electron microscopy indicated that, dependent on the stage of growth, the enzymes may form large protein aggregates.The molecular masses of both amine oxidase and D-amino acid oxidase in the mutant strains were identical to their respective counterparts in WT cells, indicating that both proteins were correctly assembled and active in the cytosol.
    Additional Material: 9 Ill.
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  • 91
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    Yeast 6 (1990), S. 1-29 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Additional Material: 10 Ill.
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  • 92
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    Yeast 6 (1990), S. 35-43 
    ISSN: 0749-503X
    Keywords: Hanseula polymorpha ; Candida boidinii ; peroxisomes ; peroxisomal membrane proteins ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have the substructure and polypeptide composition of the peroxisomal membranes in two methylotrophic yeasts in relation to different growth conditions. The results obtained that no significant ultrastructural differences existed between the membranes of variously grown cells.The presence of specific peroxisomal membrane proteins (PMPs) was studied biochemically. On sodium dodecyl sulphate-polyacrylamide gels of purified microbody membranes isolated from methanol-grown Hansenula polymorpha, prominent proteins bands were observed at 22, 31, 35, 42, 49 and 51 kD. These proteins were also present when the cells were grown in media containing ethanol and/or ethylamine. Apart from these, several other PMPs were specifically induced under these conditions, namely 24, 29, 37 and 62 kD proteins. The polypeptide composition of peroxisomal membranes from H. polymorpha was compared with that of another methylotroph. Candida biodinii. In the latter organism a specific PMP with a molecular weight of 23 kD was induced during growth on D-alanine instead of ammonium sulphate as the nitrogen source.
    Additional Material: 8 Ill.
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  • 93
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    Yeast 6 (1990) 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 94
    ISSN: 0749-503X
    Keywords: ARS elements ; shuttle vectors ; DNA replication ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have undertaken a search for autonomously replicating (ARSs) from Kluvermyces lactis chromosomal DNA able to sustain plasmid replication in K. lactis and in Saccharomyces cerevisiae. The discovery of such sequences might be interesting for the comparison of ARSs from different sources and possibly useful for the construction of multivalent vectors.HindIII fragments from K. lactis chromosomal DNA were inserted in the YIp5 plasmid (lacking an origin of replication) and the resulting chimaeric plasmids were selected for the ability to transform S. cerevisiae. Four plasmids were identified and further analysed. Two contained the same 1.8 kb K. lactis fragment and transformed both K. lactis and S. cerevisiae with the same efficiency and stability, whereas the third transformed only S. cerevisiae and the fourth transformed K. lactis with a higher efficiency than S. cerevisiae. A detailed study was performed on the 1.8kb fragment which exhibited ARS function in both yeasts. The fragment was subcloned using different restriction enzymes and Bal31 exonuclease. Subclones were tested for ARS function. ARS activities in the two yeasts were localized in the same 100 bp region. Sequencing demonstrated the presence in this region of the dodecanucleotide 5′ATTTATTGTTTT 3′ differing from the ARS core consensus of S. cerevisiae only by a T insertion.A similar nucleotide sequence of S. cerevisiae only in the putative replication origin of the 2 μ-like plasmid pkD1 which stably replicates in K. lactis. Homologies with ARSs from S. cerevisiae were also found in the regions flanking the above-mentioned dodecanucleotide.
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  • 95
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    Yeast 6 (1990), S. 139-139 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Additional Material: 1 Ill.
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  • 96
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    Yeast 6 (1990), S. 141-148 
    ISSN: 0749-503X
    Keywords: Pachysolen ; ornithine carbamoyltransferase ; xylose ; yeast ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A fragment of DNA from a yeast Pachysolen tannophilus, bearing the ornithine carbamoyltransferase gene (OCTase, EC 2.1.3.3) has been cloned from a genomic library by functional complementation of the Escherichia coli OCT-negative mutant. The gene was located within the cloned segment of DNA and its coding sequence identified by DNA sequencing. This has indicated that P. tannophilus OCT gene encodes a 347 amino acid polypeptide, which shows 60% identity to the homologous Saccharomyces cerevisiae protein. The amino acid composition of its N-terminus indicates that this protein is translocated across the mitochondrial membrane. The gene can be expressed in E. coli as well as in S. cerevisiae. Comparison with other OCTases confirms a high degree of conservation among these proteins.
    Additional Material: 3 Ill.
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  • 97
    ISSN: 0749-503X
    Keywords: Maltose transport ; α-glucosidase ; yeast ; chemostat ; cell death ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: When Saccharomyces cerevisiae CBS 8066 was grown under maltose limitation, two enzymes specific for maltose utilization were present: a maltose carrier, and the maltose-hydrolysing α-glucosidase. The role of these two enzymes in the physiology of S. cerevisiae was investigated in a comparative study in which Candida utilis CBS 621 was used as a reference organism.Maltose pulses to a maltose-limited chemostat culture of S. cerevisiae resulted in ‘substrate-accelerated death’. This was evident from: (1) enhanced protein release from cells: (2) excretion of glucose into the medium; (3) decreased viability. These effects were specific with respect to both substrate and organism: pulses of glucose to maltose-limited cultures of S. cerevisiae did not result in cell death, neither did maltose pulses to maltose-limited cultures of C. utilis. The maltose-accelerated death of S. cerevisiae is most likely explained in terms of an uncontrolled uptake of maltose into the cell, resulting in an osmotic burst. Our results also provide evidence that the aerobic alcoholic fermentation that occurs after pulsing sugars to sugar-limited cultures of S. cerevisiae (short-term Crabtree effect) cannot solely be explained in terms of the mechanism of sugar transport. Both glucose and maltose pulses to maltose-limited cultures triggered aerobic alcohol formation. However, glucose transport by S. cerevisiae occurs via facilitated diffusion, whereas maltose entry into this yeast is mediated by a maltose/proton symport system.
    Additional Material: 8 Ill.
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  • 98
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    Yeast 6 (1990) 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 99
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    Yeast 6 (1990), S. 193-204 
    ISSN: 0749-503X
    Keywords: Kluyveromyces lactis ; alcohol dehydrogenase ; gene regulation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have studied the alcohol dehydrogenase (ADH) system in the yeast Kluyvefromyces lactis. Southern hybridization to the Saccharomyces cerevisiae ADH2 gene indicates four probable structural ADH genes in K. lactis. Two of these genes have been isolated from a genomic bank by hybridization to ADH2. The nucleotide sequence of one of these genes shows 80% and 50% sequence identity to the ADH genes of S. cerevisiae and Schizosaccharomyces pombe respectively. One K. lactis ADH gene is preferentially expressed in glucose-grown cells and, in analogy to S. cerevisiae, was named K1ADH1. The other gene, homologous to K1ADH1 in sequence, shows an amino-terminal extension which displays all of the characteristics of a mitochondrial targeting presequence. We named this gene K1ADH3. The two genes have been localized on different chromosomes by Southern hybridization to an orthogonal-field-alternation gel electrophoresis-resolved K. lactis genome. ADH activities resolved by gel electrophoresis revealed several ADH isozymes which are differently expressed in K. lactis cells depending on the carbon source.
    Additional Material: 7 Ill.
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  • 100
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; cell cycle regulation ; CDC28 gene ; rho- mutation ; chromosome loss ; plasmid maintenance ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The cdc28-srm mutation in Saccheromyces cerevisiae decreases spontaneous and induced mitochondrial rhomutability and the mitotic stability of native chromosomes and recombinant circular minichromosomes. The effects of cdc28-srm on the genetic stability of cells support the hypothesis that links cell cycle regulation in yeast to changes in chromatin organization dependent on the start gene CDC28 (Hayles and Nurse, 1986).
    Additional Material: 8 Ill.
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