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  • Life and Medical Sciences  (29)
  • Mitochondria  (5)
  • Corynebacterium glutamicum  (4)
  • 1
    ISSN: 1432-0983
    Keywords: Senescence ; Plasmid ; Neurospora ; Mitochondria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Several field-collected strains of Neurospora crassa from the vicinity or Aarey, Bombay, India, are prone to precocious senescence and death. Analysis of one strain, Aarely-1e, demonstrated that the genetic determinants for the predisposition to senescence are maternally inherited. The senescence-prone strains contain a 7-kb, linear, mitochondrial DNA plasmid, maranhar, which is not present in long-lived isolates from the same geographical location. The maranhar plasmid has inverted terminal repeats with protein covalently bound at the 5′ termini. Molecular hybridization experiments have demonstrated no substantial DNA sequence homology between the plasmid and the normal mitochondrial (mtDNA) and nuclear genomes of long-lived strains of N. crassa. Integrated maranhar sequences were detected in the mtDNAs of two cultures derived from Aarey-1e, and mtDNAs with the insertion sequences accumulated during subculturing. Nucleotide sequence analysis of cloned fragments of the two insertion sequences demonstrates that that they are flanked by long inverted repeats of mtDNA. The senescence syndrome of the maranhar strains, and the mode of integration of the plasmid, are reminiscent of those seen in the kalilo strains of N. intermedia. Nonetheless, there is no detectable nucleotide sequence homology between the maranhar and kalilo plasmids.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0983
    Keywords: Gelasinospora ; Neurospora ; Plasmid ; Mitochondria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have determined the complete nucleotide sequence of a linear mitochondrial plasmid from a natural isolate of a homothallic species ofGelasinospora. The plasmid genome is 8231 by long. It carries terminal inverted repeats of 1137 bp. Extending inwards from the terminal repeats are two long open reading frames coding for putative proteins with similarity to DNA and RNA polymerases. These are separated by a short intergenic region. The plasmid sequence shows remarkable similarity to that of theNeurospora intermedia senescence-plasmid kalilo. Overall the two plasmids have a similar genetic organization and are clearly homologous at the sequence level. The main differences are in the intergenic region and in the terminal repeats.
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  • 3
    ISSN: 1617-4623
    Keywords: Corynebacterium glutamicum ; Diaminopimelate-lysine anabolic pathway ; Heterologous complementation ; Homologous expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We utilized diaminopimelate-lysine mutants of Escherichia coli K12 to clone the genes specifically involved in the Corynebacterium glutamicum diaminopimelate-lysine anabolic pathway. From a cosmid genomic bank of C. glutamicum strain AS019, we isolated cosmids pSM71, pSM61 and pSM531, that are respectively able to complement dapA/dapB, dapD, and lysA mutants of E. coli. DNA hybridization analysis indicates that these complementing genes are located on the chromosome of C. glutamicum in at least three separate transcription units. Subcloning of parental cosmids in dapA, dapD, and lysA mutants of E. coli localized these genes, respectively, within 1.4, 3.4, and 1.8 kb fragments, cloned in an E. coli/C. glutamicum shuttle vector. Enzymatic analysis in C. glutamicum identified the dapA-complementing gene as l-2,3-dihydrodipicolinate synthetase (dapA), and the lysA-complementing gene as meso-diaminopimelate decarboxylase (lysA). In contrast, complementation of E. coli dapD8, presumably lacking L-Δ1-tetrahydrodipicolinate synthetase (dapD), led us to clone a diaminopimelate-lysine anabolic gene of C. glutamicum which does not exist in E. coli: meso-diaminopimelate dehydrogenase. Although meso-diaminopimelate is crucial in lysine formation and in cell wall biosynthesis, expression of the genomic copies of the cloned genes, which encode activities involved at key branching points of the diaminopimelate-lysine pathway of C. glutamicum, appears constitutive with regard to the addition of diaminopimelate and/or lysine during cell growth.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1617-4623
    Keywords: Corynebacterium glutamicum ; lysA promoter(s) ; Weak expression ; Full expression ; Bacterial evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Sequence analysis localized the lysA gene of Corynebacterium glutamicum strain AS019 within a 1.35 kb open reading frame, potentially encoding a 445 amino acid product. Immediately downstream from this gene we found a potential ρ-independent transcription terminator, while the 5′ flanking region (300 bp) harbors unusual topological and structural features, located in the vicinity of a potential ribosome binding site. Within this upstream region, enzymatic and genetic analyses indicated the occurrence of a promoter responsible for significant, although weak, expression of the encoded enzymatic activity. The same significant expression level was observed with a plasmid harboring an additional 0.5 kb of genomic information upstream from lysA, while its full expression apparently requires 2 kb of additional genomic information located immediately upstream from the cloned gene. The upstream sequence requirement apparently associated with the full expression of the lysA gene of C. glutamicum shows some similarity with the Escherichia coli system.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1617-4623
    Keywords: Corynebacterium glutamicum ; Phosphoenolpyruvate carboxylase ; ppc gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The ppc gene of Corynebacterium glutamicum encoding phosphoenolpyruvate (PEP) carboxylase was isolated by complementation of a ppc mutant of Escherichia coli using a cosmid gene bank of chromosomal c. glutamicum DNA. By subsequent subcloning into the plasmid pUC8 and deletion analysis, the ppc gene could be located on a 3.3 kb SalI fragment. This fragment was able to complement the E. coli ppc mutant and conferred PEP carboxylase activity to the mutant. The complete nucleotide sequence of the ppc gene including 5′ and 3′ flanking regions has been determined and the primary structure of PEP carboxylase was deduced. The sequence predicts a 919 residue protein product (molecular weight of 103154) which shows 34% similarity with the respective E. coli enzyme.
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 220 (1989), S. 113-120 
    ISSN: 1617-4623
    Keywords: Neurospora ; Senescence ; Mitochondria ; Plasmid ; Kalilo
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The linear autonomous form of kalilo DNA (previously called AR-kalDNA) is shown to be resident within mitochondria rather than nuclei, as had been suggested by previous experiments. This form has been renamed mtAR-kalDNA, to signify its mitochondrial location. Experiments are described that illustrate the inheritance and somatic transmission patterns of the mitochondrial kalilo plasmid and the mitochondrial inserted form of kalilo DNA (mtlS-kalDNA). Progeny of a cross with a pre-senescent subculture as the female parent inherited mtAR-ka1DNA only; mtIS-kalDNA was not transmitted sexually. During somatic propagation of the ascospore cultures, novel kalilo DNA inserts appeared and most of them persisted until death. We propose that these inserts originated from de novo integration of mtAR-kalDNA into the mitochondrial DNA. In two of the ascopore-derived series analyzed, the first inserts detected were seen only transiently and inserts appearing subsequent to the transient inserts were retained until death. We propose that these enduring inserts originated either from rearrangements of the transient inserts or from novel integration events, either from mtAR-kalDNA or from transposition of the transient inserts.
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 237 (1993), S. 177-186 
    ISSN: 1617-4623
    Keywords: Senescence ; Linear plasmids ; Circular plasmids ; Neurospora ; Mitochondria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A sample of 171 natural isolates of Neurospora crassa and Neurospora intermedia was tested for senescence. Of these, 28 strains senesced within the duration of the experiment. These senescent strains, together with a selection of nonsenescent strains, were examined for the presence of mitochondrial plasmids. This was done by digesting mitochondrial DNA preparations with proteinase K, and running these samples on agarose gels. Most of the strains examined, both senescent and nonsenescent, contained plasmids, many of them new. Some new plasmids were linear, as inferred from their resistance to 5′ exonuclease and sensitivity to 3′ exonuclease. New circular plasmids were also found. Some strains carry several plasmids, and mixtures of circular and linear elements were common. A cross-homology study was performed on a sample of plasmid-bearing strains, and several cases of apparent relatedness were found, some between strains from distant geographical locations. Linear plasmids homologous to the maranhar linear senescence plasmid were quite common. A new member of the LaBelle circular plasmid homology group was found. In the sample tested for homology, no strains contained elements related to the kalilo linear senescence plasmid. The relationship of the new plasmids to senescence is not known. In addition to plasmid monomers, several different types of derivatives were found. The kalilo linear plasmid was found to occur in linear and circular forms of low mobility, presumed to be giant concatamers, and, in some strains, variant sibling structures and ladders of short derivatives were found. Circular plasmids also gave rise to extensive ladders on electrophoresis, probably representing different relaxation states and head-to-tail concatameric series. Some such forms migrated more slowly than mitochondrial DNA. One unique type of plasmid modification observed was a pair of linear elements that had apparently arisen de novo which showed homology to a circular plasmid.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 249 (1995), S. 25-36 
    ISSN: 1617-4623
    Keywords: Neurospora ; Plasmids ; Mitochondria ; Recombination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A strain of Neurospora intermedia from China contains five prominent extragenomic mitochondrial plasmids: three linear elements called zhisi plasmids, and two circular plasmids, Harbin-1 and -2. In one subculture, levels of four plasmids (all three zhisis and Harbin-1) fell to undetectable values and two novel linear plasmids appeared, Harbin-L and -L2, as well as a new small circular plasmid, Harbin-0.9. Cross-hybridization of restriction fragments and DNA sequencing showed that the Harbin-L plasmid was composed of parts of the circular Harbin-1 plasmid and of one of the linear zhisi plasmids. A model is presented in which the Harbin-1 and zhisi plasmids are present within the same mitochondrion, and crossovers at two separate 7 by sites of sequence identity effectively insert part of the circular Harbin-1 DNA into a zhisi linear plasmid, simultaneously deleting part of the zhisi element. The small plasmid Harbin-0.9 is a fragment of the Har-1 plasmid, and seems to be another product of the recombination process that created Har-L. Recombination of this type could have contributed to the wide array of mitochondrial plasmids found in natural populations of Neurospora.
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Antonie van Leeuwenhoek 67 (1995), S. 221-227 
    ISSN: 1572-9699
    Keywords: lysine biosynthesis ; aspartate-derived amino acids ; oxaloacetate decarboxylase ; pyruvate kinase ; Corynebacterium glutamicum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Oxaloacetate (OAA) decarboxylase (E.C. 4.1.1.3) was isolated fromCorynebacterium glutamicum. In five steps the enzyme was purified 300-fold to apparent homogeneity. The molecular mass estimated by gel filtration was 118 ± 6 kDa. SDS-PAGE showed a single subunit of 31.7 KDa, indicating an α4 subunit structure for the native enzyme. The enzyme catalyzed the decarboxylation of OAA to pyruvate and CO2, but no other α-ketoacids were used as substrate. The cation Mn2+ was required for full activity, but could be substituted by Mg2+, Co2+, Ni2+ and Ca2+. Monovalent ions like Na+, K+ or NH 4 + were not required for activity. The enzyme was inhibited by Cu2+, Zn2+, ADP, coenzyme A and succinate. Avidin did not inhibit the enzyme activity, indicating that biotin is not involved in decarboxylation of OAA. Analysis of the kinetic properties revealed a K m for OAA of 2.1 mM and a K m of 1.2 mM for Mn2+. The V max was 158 µmol of OAA converted per min per mg of protein, which corresponds to an apparent k cat of 311 s−1.
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  • 10
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
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