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  • Replicative transformation  (2)
  • Salt tolerance  (1)
  • osmotically regulated gene expression  (1)
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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Current genetics 24 (1993), S. 114-121 
    ISSN: 1432-0983
    Keywords: Pleurotus astreatus ; Origin of replication ; Nucleotide sequence ; Replicative transformation ; Recombinant plasmid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Replicative plasmids generated through in-vivo recombination have been identified among transformants of the fungus Pleurotus ostreatus. In addition to sequences from a standard selection vector (pAN7-1), these recombinant plasmids contain recombined sequences of chromosomal origin conferring replicative potential upon the vector. One such recombined sequence, an 1148-bp insert into plasmid pP01, has been characterized. This sequence has been analyzed for secondary structural features as well as for consensus sites affiliated with origins of replication (ori) in other eukaryotic systems. The 1148-bp insert lacks an ORF and does not contain an acceptable match to the commonly identified 11-bp ars consensus sequence (A/TTTTATA/GTTTA/T) for autonomous replication in the yeast Saccharomyces cerevisiae. The analysis, however, revealed a cluster of three hairpin-loop-forming subsequences with individual ΔG25°C free energy values of-7.6,-6.4 and-5.2 kcal mol-1. Also found were two 7-bp analogues to centromere-affiliated sequences recognized in other fungi, as well as several putative gyrase recognition sites comparable to the 9-bp S. cerevisiae/E. coli gyrase-binding consensus sequence. Sequences comparable to the ori of the yeast 2-μm plasmid or to various sequences associated with ori of yeast/fungal mitochondrial DNAs (mtDNA) were not present in the 1148-bp insert. Replication of pP01 appears rather to involve a replication of chromosomal derivation devoid of an ars-type consensus.
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Current genetics 29 (1996), S. 130-135 
    ISSN: 1432-0983
    Keywords: Salt stress ; Differential gene expression ; Salt tolerance ; Aspergillus nidulans
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Differentially expressed cDNA clones were isolated from salt-adaptedAspergillus nidulans (FGSC #359). Poly (A)+ RNA from adapted mycelia was used to construct a λ Uni-ZAP cDNA library. The library was screened with mixed subtracted cDNA probes. Three-hundred and fifty-seven positive plaques were isolated in the primary screening. Sixty-two randomly selected plaques were purified and placed into eight different cross-hybridization groups. A representative cDNA from each group was used to study expression under unadapted, salt-adapted and salt-shock conditions. These clones, representing eight different genes, displayed enhanced expression under salt stress. Exploratory nucleotide sequencing was performed, and the predicted amino-acid sequence was compared with known gene sequences in the data-bank. Five of the cDNA clones were identified as a mitochondrial (mt) ATPase β subunit, a mt ATPase subunit 9, a mt transport protein, a ubiquitin-extension protein and a ribosomal protein. Three cDNA clones could not be identified due to lack of adequate homology with known sequences. These results suggest that at least five genes with known function in cellular processes like ATP generation and protein synthesis, and three other genes of unknown identity, are greatly induced in salt-adapted conditions.
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Current genetics 22 (1992), S. 53-59 
    ISSN: 1432-0983
    Keywords: Pleurotus ; Replicative transformation ; Recombinant plasmids ; DNA methylation ; Plasmid recovery
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A transformation system employing selectable resistance to hygromycin B has been developed for the mushroom-forming fungus, Pleurotus ostreatus. Vector pAN7-1, a commonly used non-replicative vector for integrative transformation in fungi, yielded 5–46 resistant colonies per μg of DNA per 107 viable protoplasts. Southern blot analysis of certain transformants revealed unexpected replicative plasmids containing pAN7-1 sequences, but modified for size, methylation and restriction enzyme pattern when compared to the initial transforming vector. Two such replicative derivatives of pAN7-1 have been rescued from P. ostreatus by cloning into Escherichia coli. Rescued plasmids have been used to probe DNA from untransformed P. ostreatus in an effort to identify fungal sequences that recombined in vivo with pAN7-1 to form replicative plasmids. Such replicative sequences have been localized in high molecular weight (chromosomal) DNA of wild-type P. ostreatus. Transformation has been obtained for P. ostreatus using a rescued plasmid, thereby confirming the role of this recombinant plasmid as a shuttle vector.
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  • 4
    ISSN: 1573-5028
    Keywords: Nicotiana tabacum L. ; tobacco ; ABA ; ethylene ; salt ; osmotically regulated gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A Nicotiana tabacum gene encoding the basic PR-like protein osmotin was isolated and characterized. The gene is derived from the N. sylvestris parent of N. tabacum. In cell suspension cultures of tobacco, the osmotin gene was shown to be transcriptionally activated by treatment with ABA. Transcriptional activation of the osmotin promoter was further investigated in transformed plants carrying copies of a fusion of the cloned promoter to the β-glucuronidase reporter gene. In these plants, the osmotin promoter is transcriptionally activated by the hormones ABA and ethylene. The sensitivity of the osmotin promoter to ABA applied exogenously decreased with age in both roots and shoots of young seedlings. NaCl shock also activated the promoter in plant tissues. The osmotin promoter is much more active in root tissues than in shoot tissues.
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