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  • Digitale Medien  (17)
  • 1990-1994
  • 1985-1989  (9)
  • 1960-1964
  • 1955-1959  (8)
  • 1945-1949
  • 1930-1934
  • 1920-1924
  • 1870-1879
  • 1986  (9)
  • 1958
  • 1955  (8)
  • 1876
  • 1872
  • gene expression
  • somatic hybridization
Materialart
  • Digitale Medien  (17)
Erscheinungszeitraum
  • 1990-1994
  • 1985-1989  (9)
  • 1960-1964
  • 1955-1959  (8)
  • 1945-1949
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Jahr
  • 1
    ISSN: 1573-4919
    Schlagwort(e): cell cycle ; gene expression ; oncogenes
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie , Medizin
    Notizen: Summary A number of genes and cDNA sequences (including at least four oncogenes) are known to be expressed in a cell cycle-dependent manner, i.e. the levels of specific mRNAs vary with the phases of the cell cycle. In order to explore the significance of some of these sequences in the mitogenic response, we have investigated the expression of 8 cell cycle-dependent sequences (plus two control sequences, not expressed in a cell cycle-dependent manner) under a variety of conditions. These conditions included cells of different types, from different species, stimulated to proliferate by different mitogens. The genes (or sequences) studied included five cDNA clones whose sequences are preferentially expressed in early G1, i.e. two cDNA clones inducible by platelet-derived growth factor (JE-3 and KC-1), and three cDNA clones inducible by serum (2A9, 2F1, 4F1); and three oncogenes (c-myc, c-rasHa and p53) whose expression is known to be cell cycle-dependent. All of the tested genes, except 2A9, c-rasHa and the control genes, are expressed in a cell cycle-dependent manner in human peripheral blood mononuclear cells stimulated by phytohemagglutinin and in serum-stimulated mouse and Syrian hamster fibroblasts. The inducibility of these genes by different mitogens in cells of different types and from different species strongly suggests that these genes play a role in cell cycle progression. This conclusion is further supported by the known structural and functional similarities between cell-cycle dependent genes, oncogenes and genes coding for cell-cycle related molecules.
    Materialart: Digitale Medien
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  • 2
    Digitale Medien
    Digitale Medien
    Springer
    Bioscience reports 6 (1986), S. 621-631 
    ISSN: 1573-4935
    Schlagwort(e): brown adipose tissue ; thermogenin ; uncoupling protein ; gene expression ; adrenergic effects
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie
    Notizen: Abstract By the use of an earlier characterised cDNA clone, CIN-1, corresponding to a sequence of the mRNA coding for the brown-fat specific “uncoupling” protein, thermogenin, the amount of thermogenin mRNA found in the brown adipose tissue of mice was quantitatively investigated under different physiological and pharmacological conditions. It was found that a 4 hr cold stress led to a 7-fold increase in the amount of thermogenin mRNA; injection of norepinephrine had a significant but smaller effect. Most notably, isoprenaline (β-agonist) and phenylephrine (α-agonist) had in themselves no effect, but when injected together were able to increase the mRNA level synergistically. In 4 hr cold-stressed mice, norepinephrine, isoprenaline and cholera toxin could all further potentiate the effect of the cold stress itself on the mRNA level. Insulin and the glucocorticoid dexamethasone both had weak stimulatory effects on the mRNA level. It is concluded that an increase in intracellular cAMP levels is a necessary and perhaps sufficient stimulus for the increase in thermogenin gene expression. However, at least underin vivo conditions, this increase requires stimulation of both α- andβ-adrenergic pathways.
    Materialart: Digitale Medien
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  • 3
    Digitale Medien
    Digitale Medien
    Springer
    Plant molecular biology 7 (1986), S. 51-61 
    ISSN: 1573-5028
    Schlagwort(e): Soybean ; Rhizobium ; nitrogen fixation ; gene expression
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary Nodulins are organ-specific plant proteins induced during symbiotic nitrogen fixation. Nodulins play both metabolic and structural roles within infected and uninfected nodule cells. In soybean, several nodulin genes, coding for abundant nodulins, have been identified and isolated. Structural analysis of some of these genes has revealed their possible mode of regulation and the subcellar location of the protein product. Studies of ineffective symbiosis based on cultivar-strain genotype differences suggested that both partners influence the expression of nodulin genes. Concomitant with nodule organogenesis, the Rhizobium undergoes substantial differentiation leading to the accumulation of nodule-specific bacterial proteins, bacteroidins. The major structural alteration occuring in the infected cell is the formation of a membrane enclosing the bacteroid (peribacteroid membrane). A number of nodulins are specifically targetted to this membrane during endosymbiosis. The induction of nodulins and bacteroidins leads to the formation of an effective nodule. Nodulin genes can be induced in vitro by factors derived from nodules suggesting that trans-activators may be involved in derepression of the host genes necessary for Rhizobium-legume symbiosis.
    Materialart: Digitale Medien
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  • 4
    ISSN: 1573-5028
    Schlagwort(e): Agrobacterium rhizogenes ; Solanum tuberosum ; T-DNA ; gene expression ; tuberisation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary Expression of TL-DNA from Agrobacterium rhizogenes plasmid pRi 1855 was examined in a transformed derivative of Solanum tuberosum cv. Desiree, D9X8a. Northern blot analysis identified at least nine TL-DNA coded transcripts in roots, shoots and tubers but their relative abundance differed within and between organs. This revealed a distinctive pattern of organ specified differential expression. Grafting experiments showed that the abnormal shape of tubers of transformed potato was probably determined by TL-DNA products synthesised within the tuber and not by diffusable products synthesised in other parts of the plant. The abundance of at least one transcript, tr5, was probably determined by culture conditions. Implications for functions and control of expression of Ri TL-DNA genes are discussed. It is suggested that Ri TL-DNA provides a convenient and extensive set of model genes to study variation and stability of expression of linked foreign genes introduced into plants.
    Materialart: Digitale Medien
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  • 5
    Digitale Medien
    Digitale Medien
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 116-124 
    ISSN: 0887-3585
    Schlagwort(e): translational repressor ; in vitro transcription-translation ; gene expression ; protein purification ; Chemistry ; Biochemistry and Biotechnology
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Medizin
    Notizen: The bacteriophage T4 translational represor regA protein has been purified from an overproducing strain, and its activity has been studied in simple in vitro protein synthesis reactions. RegA protein was found to inhibit the translation of T4 genes 44, 45, and ORF45-1 in a concentration-dependent fashion. Expression of two other T4 genes which are insensitive to regA protein in vivo, genes 32 and 43, was unaffected by the presence of regA protein. Specific inhibition of synthesis of genes 44, 45, and ORF 45-1 proteins was achieved with 5-20 μM concentrations of regA protein, without the addition of any other T4 encoded proteins or cofactors. When in vitro protein synthesis was performed in two steps, uncoupling translation from transcription, regA protein had an inhibitory effect regardless of whether it was added at the initiation of transcription or only at the translation step. This indicates that regA protein functions during the translation step of protein synthesis in vitro in agreement with previous in vivo studies of regA protein.
    Zusätzliches Material: 7 Ill.
    Materialart: Digitale Medien
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  • 6
    ISSN: 0730-2312
    Schlagwort(e): tumor metastasis ; gene expression ; oncogenes ; virus antigens ; glycoproteins ; Life and Medical Sciences ; Cell & Developmental Biology
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Biologie , Chemie und Pharmazie , Medizin
    Notizen: A metastatic variant cell subline of the Abelson virus-transformed murine large lymphoma/lymphosarcoma RAW 117 has been selected in vivo ten times for liver colonization. Highly metastatic subline RAW117-H10 forms greater than 200 times as many gross surface liver tumor nodules as the parental line RAW117-P. Analysis of cellular proteins and glycoproteins indicates reduced expression of murine Moloney leukemia virus-associated p15, p30, and gp70, and increased expression of a sialoglycoprotein, gp150, in the highly metastatic H10 cells. Northern analyses of oncogene expression suggested that mRNA of various oncogenes was expressed equally or not expressed in the RAW117 cells of differing metastatic potential. Differential gene expression was examined using a cDNA library of 17,600 clones established from poly A+ mRNA isolated from H10 cells. The cDNA library was screened by the colony hybridization technique using probes made from both RAW117-P and -H10 cells. Approximately 99.5% of these cDNA clones were expressed identically in P and H10 cells. Of the few differentially expressed cDNA clones (approx. 150/17,600), one-half of these were identified as Moloney leukemia virus sequences in a separate probing with a radiolabeled Moloney leukemia virus probe. The remainder of the differentially expressed mRNA detected by colony hybridization of the cDNA library were expressed at higher levels (approx. 1/6) or lower levels (approx. 1/3) in the highly metastatic H10 cells.
    Zusätzliches Material: 3 Ill.
    Materialart: Digitale Medien
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  • 7
    Digitale Medien
    Digitale Medien
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 7 (1986), S. 149-158 
    ISSN: 0192-253X
    Schlagwort(e): macronucleus ; gene expression ; cell union ; Life and Medical Sciences ; Genetics
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Biologie
    Notizen: We report here the presence of N6-methyladenine (MeAde) in the macronuclear DNA (maDNA) of Blepharisma japonicum vegetative cells. We have further investigated the relationship between DNA methylation and cell union in cells activated for conjugation. Such activation was induced by treating cells of mating type I with complementary gamone 2. We found a reduction of about 24% of MeAde content in gamone-treated cells ready for cell union. First indications of the presence and reduction of MeAde content came from electrophoresis of maDNA digested by appropriate restriction endonucleases. Chromatographic determination of the amount of methylated base by HPLC substantiated these observations. In vegetative cells, 1.576 ± 0.02% of total adenine was found to be methylated as opposed to 1.193 ± 0.04% in activated cells. The HPLC analysis of maDNA also revealed a peak with a retention time corresponding to that of 5-hydroxymethyluracil, already found in some species of dinoflagellates. In that gamone treatment is correlated with a differential gene expression (indicated by a differential RNA and protein synthesis), our results suggest that there is a relationship between macronuclear genome activation and demethylation of maDNA. This is the first report of a correlation between gene activation and adenine demethylation in a eukaryotic organism.
    Zusätzliches Material: 4 Ill.
    Materialart: Digitale Medien
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  • 8
    Digitale Medien
    Digitale Medien
    New York, NY [u.a.] : Wiley-Blackwell
    Archives of Insect Biochemistry and Physiology 3 (1986), S. 59-73 
    ISSN: 0739-4462
    Schlagwort(e): gene expression ; chorion ; homology ; cockroach ; oothecin ; juvenile hormone ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Biologie
    Notizen: Control of specific protein synthesis by juvenile hormone during sexual maturation and reproduction is being studied in the left colleterial gland of the cockroach Periplaneta americana. Rates of specific oothecin synthesis and oothecin mRNA concentrations have been measured during sexual maturation and the effects of allatectomy and administration of juvenile hormone determined. The heterogeneity of the oothecins has been characterized and found to be due probably to a multiplicity of structural genes. The complete primary structure of the major oothecin has been deduced from the sequences of cloned cDNAS. It can be divided into a central region and two flanking arms that share sequence features. The structures of the oothecins show strong homologies with the silkmoth chorion proteins.
    Zusätzliches Material: 6 Ill.
    Materialart: Digitale Medien
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  • 9
    Digitale Medien
    Digitale Medien
    New York, NY [u.a.] : Wiley-Blackwell
    Archives of Insect Biochemistry and Physiology 3 (1986), S. 119-132 
    ISSN: 0739-4462
    Schlagwort(e): Ecdysone ; cuticle proteins ; gene expression ; Drosophila ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Biologie
    Notizen: The deposition and apolysis of insect cuticles have long been known to be regulated by ecdysone. Unsclerotized, chitin-containing procuticles contain evolutionarily conserved, hydrophobic proteins that are soluble in solutions of denaturing agents. The pupal procuticle of Drosophila is deposited by larval and imaginal epidermis starting 9 h after puparium formation when the ecdysone titer is low. Initially, a set of low molecular weight proteins (less than 25,000 daltons; low molecular weight pupal cuticle proteins = S-PCPs) is synthesized. However, about the time of pupation, synthesis of S-PCPs ceases, and high molecular weight proteins (greater than 50,000 daltons; H-PCPs) are synthesized. In vitro experiments indicate that the initial formation of the procuticle with synthesis of the S-PCPs requires a pulse of hormone followed by withdrawal (6 h with 20-hydroxyecdysone, 1 μg/ml). The switch from synthesis of S-PCPs to H-PCPs is facilitated by a second, short pulse of 20-hydroxyecdysone (0.1 μg/ml, 3 h). Ultrastructural localization demonstrates that the S-PCPs are located only in the external lamellae of the procuticle, while the H-PCPs are present only in internal lamellae. Developmental analyses with cloned genes indicate that cuticle protein genes are expressed during only one stage of Drosophila development. Some of the genes encoding S-PCPs are limited in their expression to larval (posterior) or imaginal (anterior) epidermis. Preliminary molecular analyses of the larval and pupal cuticle protein genes indicate that they are organized in different ways. For example, four larval genes exist in a cluster with divergent transcription, and one PCP gene, PCP-GART, is located within an intron of a “housekeeping” gene.
    Zusätzliches Material: 5 Ill.
    Materialart: Digitale Medien
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  • 10
    ISSN: 1573-5060
    Schlagwort(e): protoplasts ; protoclonal variation ; somatic hybridization ; top-fruit trees ; woody ornamentals
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Summary Until recently, temperate fruit trees and woody ornamentals have been regarded as recalcitrant to biotechnological breeding approaches based on protoplasts. This however should no longer be the case, as procedures are now available, not only for the regeneration of complete plants from protoplasts of various tissues of such species, but also for the exploitation of protoplast technology for their genetic manipulation. This paper will examine the recent advances and state of the art in this domain, with particular attention to the use of protoplast technology as a novel tool in the breeding of rosaceous top-fruit tree species and woody ornamentals. Problems and their solutions within the context of regenerating plants from isolated protoplasts of stone (Prunus spp.), pome (Pyrus spp., Malus spp.) and small (Rubus spp.) fruits, and of several shrubby ornamental genotypes (Lonicera spp., Weigela spp., Forsythia spp., Cotoneaster spp.) will be addressed. Interspecific (Prunus spinosa + Prunus cerasifera) and intergeneric (Forsythia spp. + Syringa spp.) somatic hybridization within this group of species, as well as the use of protoplasts for host/pathogen interaction studies (Pyrus/Erwinia amylovora) will also be discussed.
    Materialart: Digitale Medien
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  • 11
    Digitale Medien
    Digitale Medien
    Springer
    Euphytica 85 (1955), S. 217-233 
    ISSN: 1573-5060
    Schlagwort(e): crop improvement ; alien gene transfer ; progeny analysis ; somatic hybridization
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Summary In recent years, the rapid development of somatic cell genetics has made possible the transfer of alien genes over wide taxonomic distances by somatic hybridization. In this review, the potential of somatic hybridization in the breeding of crops within the Brassicaceae, Fabaceae, Poaceae and Solanaceae is discussed. It is evident from these studies that many hybrids, either symmetric or asymmetric, which are fertile have the potential to be used as a bridge between the alien species and the crop. Progeny analysis of some hybrid combinations also reveals intergenomic translocations which may lead to the introgression of the alien genes. Furthermore, fusion techniques enable the resynthesis of allopolyploid crops to increase their genetic variability and to restore ploidy level and heterozygosity after breeding at reduced ploidy level in polyploid crops.
    Materialart: Digitale Medien
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  • 12
    ISSN: 1573-5060
    Schlagwort(e): microprotoplast fusion ; partial genome transfer ; monosomic additions ; kanamycin resistance ; β-glucuronidase ; gene expression ; potato
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Summary Various aspects of a microprotoplast fusion technique and the strategies followed for intergeneric partial genome transfer (one or a few chromosomes) and alien genes from sexually-incongruent donor species to recipient species are described. The essential requirements of the microprotoplast fusion technique are the induction of micronuclei at high frequencies, as well as the isolation and enrichment of sub-diploid microprotoplasts in donor species, efficient fusion of the donor microprotoplasts with normal recipient protoplasts and stable regeneration of plants from fusion products. The results on the production of microprotoplast hybrid plants between the transformed donor lines of Solanum tuberosum and Nicotiana Plumbaginifolia carrying various genetic markers, and a recipient line of Lycopersicon peruvianum or Nicotiana tabacum, and on the transfer and expression of alien genes (kanamycin resistance, β-glucuronidase) are presented. The data obtained on microprotoplast hybrid plants between S. tuberosum and L. peruvianum showed that many of the hybrids contained one potato chromosome carrying nptII and GUS, and 24 or 48 L. peruvianum chromosomes (monosomic additions), and that they were male-and female-fertile. Various applications of chromosome transfer by this technique, especially for economically-important traits (e.g. disease or stress resistance) from sexually-incompatible wild species, for construction of chromosome-specific DNA libraries through microdissection and microcloning of chromosomes, or by flow-sorting of chromosomes for genome analysis, are discussed.
    Materialart: Digitale Medien
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  • 13
    ISSN: 1573-5060
    Schlagwort(e): Agrobacterium ; Brassica napus ; CaMV 35S promoter ; mas promoter ; gene expression ; risk assessment
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Summary Gene fusions between the β-glucuronidase (GUS) reporter gene and the promoters of the cauliflower mosaic virus 35S RNA transcript (CaMV 35S) and the mannopine synthase (mas) genes were introduced into rapeseed varieties via Agrobacterium-mediated transformation. Fluorometric assay of β-glucuronidase activity indicated different expression patterns for the two promoters. In seedlings, the CaMV 35S promoter had maximum activity in the primary roots, while the mas promoter was most active in the cotyledons. Etiolated seedlings cultured in the dark showed reduced activity of the mas promoter. Before vernalization at the rosette stage, both promoters were more active in older plant parts than in younger ones. At this stage the highest activity was recorded in cotyledons. After the plants had bolted reduced promoter function was detected in the upper parts of the transformed plants. Both promoters were found to be functional in the majority of the studied organs of transgenic rapeseed plants, but the promoter activity varied considerably between the organs at different developmental stages. The ability of pollen to transfer the introduced genes to other varieties and related species (e.g. Brassica napus and Diplotaxus muralis) by cross-pollination was studied in greenhouse experiments, and field trials were carried out to estimate the distance for biologically-relevant gene dispersal. In artificial crossing, the introduced marker gene was transferable into other varieties of Brassica napus. In field trials, at a distance of 1 metre from the source of transgenic plants, the frequency of an outcrossing event was relatively high (10-3). Resistant individuals were found at 16 and 32 metres from the transgenic pollen donors, but the frequency of an outcrossing event dropped to 10-5.
    Materialart: Digitale Medien
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  • 14
    ISSN: 1573-5060
    Schlagwort(e): Solanum tuberosum ; somatic hybridization ; regeneration ; asymmetric fusion
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Summary This paper reviews investigations into the application of protoplast fusion to the genetic and agronomic improvement of potato. Fusion studies involving Solanum tuberosum are reviewed under the categories of: fusion with wild relatives, dihaploid fusion and asymmetric strategies. The selection and characterisation of putative somatic hybrid material is identified as a critical stage in the process and certain specific aspects of this technology are identified. Future prospects for the wider uptake and integration of these techniques into breeding programmes are also discussed.
    Materialart: Digitale Medien
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  • 15
    Digitale Medien
    Digitale Medien
    Springer
    Euphytica 85 (1955), S. 281-285 
    ISSN: 1573-5060
    Schlagwort(e): Lactuca sativa ; Lactuca virosa ; Lactuca tatarica ; Lactuca perennis ; Iettuce ; sexual hybridization ; embryo rescue ; somatic hybridization ; protoplast fusion
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Summary Various genes for disease resistance identified in wild Lactuca are difficult, even impossible to exploit in lettuce breeding, due to sexual incompatibility between L. sativa and wild Lactuca sp. We adapted two cellular biology techniques to overcome these interspecific barriers: in vitro embryo rescue and protoplast fusion. In vitro rescue of immature embryos was used successfully for sexual hybridization between L. sativa and L. virosa. Vigorous hybrid plants were produced between L. sativa and seven accessions of L. virosa. Protoplast fusion permitted the regeneration of somatic hybrids between L. sativa and either L. tatarica or L. perennis. Hybrids between L. sativa and L. tatarica were backcrossed to L. sativa.
    Materialart: Digitale Medien
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  • 16
    ISSN: 1573-5060
    Schlagwort(e): carotenoids ; ethylene ; gene expression ; Lycopersicon esculentum Mill. ; polygalacturonase ; pectinesterase ; phytoene synthase ; ACC oxidase
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Summary The common cultivated tomato (Lycopersicon esculentum Mill.) provides a major focus for improvement of crop quality through genetic engineering. Identification of ripening-related cDNAs has enabled the modification of specific aspects of ripening by manipulating gene expression in transgenic plants. By utilizing ‘antisense RNA’ to modify expression of ripening genes, we have inhibited the production of the cell wall-metabolising enzymes polygalacturonase and pectinesterase and created transgenic plants that contain, effectively, single, targeted mutations affecting these genes. Furthermore, this approach has been used with previously unidentified cDNA clones to enable both functional identification and manipulation of genes involved in ethylene production (ACC oxidase) and carotenoid biosynthesis (phytoene synthase). The use of antisense RNA targeted to specific genes to alter ripening phenotypes and improve commercial utility of fruit by affecting shelf-life, processing characteristics and nutritional content is discussed. We have used the extreme ripening-impaired mutant, ripening inhibitor (rin) to identify additional genes implicated in the ripening process. This approach has resulted in the cloning of several novel ripening-related mRNAs which are now being studied by antisense experiments. This may enable identification and manipulation of additional genes involved in processes such as softening, flavour and aroma generation and susceptibility to pathogens.
    Materialart: Digitale Medien
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  • 17
    ISSN: 1573-5060
    Schlagwort(e): Vicia narbonensis ; gene transfer ; gene expression ; seeds ; 2S albumin ; methionine
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Summary Epicotyl explants were co-cultivated with Agrobacterium tumefaciens EHA101 to transfer a chimeric 2S albumin gene construct carried in the binary Ti plasmid vectors pGSGLUC1 or pGA472 into the grain legume Vicia narbonensis. This gene encoding the sulphur-rich Brazil nut albumin was under the control of either the CaMV 35S promoter which permits gene expression in all organs, or the Vicia faba legumin B4 promoter which elicits seed-specific gene expression. After callus formation and selection for kanamycin resistance, somatic embryos were induced which, in the case of transformation with the vector pGSGLUC1, were screened for GUS activity. Embryos that produced GUS were in addition analysed for 2S albumin formation. Selected transgenic embryos were cloned by multiple shoot regeneration. Rooted and fertile plants were obtained by grafting transgenic shoots on the appropriate seedlings. R1 and R2 generations were raised and analysed for GUS as well as 2S albumin gene expression. Expression of the 35S promoter/2S albumin gene fusion took place in all organs of the transgenic plants including the cotyledons of seeds, whereas seed-specific gene expression was found in transformants with the legumin promoter/2S albumin gene fusion. The 2S albumin accumulated in the 2S protein fraction of transgenic seeds and its primary translation product was processed into the 9 and 3 kDa polypeptide chains. The foreign protein was localised in the protein bodies of the grain legume. Analysis of the R2 plants indicated Mendelian inheritance of the 2S albumin gene. In homozygous V. narbonensis plants the amounts of 2S albumin were twice that present in the corresponding heterozygous plants. Whereas only low level formation of the foreign protein was achieved if the gene was under the control of the 35S promoter, approximately 3.0% of the soluble seed protein was 2S albumin if seed-specific gene expression was directed by the legumin B4 promoter. Some of these transformants exhibited a three-fold increase in the methionine content of the salt-soluble protein fraction extracted from seeds.
    Materialart: Digitale Medien
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