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  • 11
    Electronic Resource
    Electronic Resource
    Regulatory elements and interactions in the Drosophila 68C glue gene cluster (1989)
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 10 (1989), S. 189-197 
    Details
    ISSN: 0192-253X
    Keywords: Drosophila ; transformation ; Glue gene ; Gene cluster ; Gene regulation ; Sgs-3 ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We reviewed studies on the developmental regulation of the 68C glue gene cluster of Drosophila melanogaster. Extensive transformation analyses of Sgs-3 have shown that four regions necessary for normal expression can be distinguished. The first ( + 10 to -50) contains the transcription start site and TATA motif. This region can be replaced functionally by corresponding sequences from the hsp70 gene, but it is sensitive to point mutations in the TATA sequence. The second region (-50 to -98) contains more than one upstream sequence that, in combination with the other elements, leads to stage and tissue-specific expression. The third region (centered at -600) contains an element that enhances transcript levels some 20-fold. The final region (between -1.65 and -2.35 kb) contains elements having modest (twofold to threefold) effects on expression, one of which is contained in the coding sequences of Sgs-7, a second member of the cluster.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/dvg.1020100308
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  • 12
    Electronic Resource
    Electronic Resource
    A haploid and a diploid cell cycle coexist in an in vitro immortalized spermatogenic cell line (1995)
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 16 (1995), S. 119-127 
    Details
    ISSN: 0192-253X
    Keywords: Germ cell line ; FACS ; mitosis ; meiosis ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have recently established a conditionally immortalized germ cell line [GC-2spd(ts)] that, at the permissive temperatures of 37°C and 32°C, is able to undergo meiosis in vitro and form round spermatids [Hofmann et al., (1994): Proc Natl Acad Sci USA 91:5533-5537]. In this report, we provide data that indicate that the GC-2spd(ts) cell line consists of two cell populations undergoing a haploid (n/2n) cell cycle and a diploid (2n/4n) cell cycle, respectively. The cells containing 2n DNA, when sorted by fluorescence-activated cell sorting, are able to reconstitute the full population of n/2n/4n DNA cells, indicating that they are able to commit to the reductive meiotic division and form haploid spermatids or to continue self-replication through a diploid cell cycle. This GC-2spd(ts) cell line provides a valuable tool to study the molecular mechanisms involved in the cellular decision between self-renewal by mitosis and commitment to meiosis. © 1995 Wiley-Liss, Inc.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/dvg.1020160205
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  • 13
    Electronic Resource
    Electronic Resource
    Development of a transformation system for the yeast Yamadazyma (Pichia) ohmeri (1994)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 10 (1994), S. 1601-1612 
    Details
    ISSN: 0749-503X
    Keywords: Pichia ; β-isopropylmalate dehydrogenase ; orotidine 5′-monophosphate decarboxylase ; genetic transformation ; gene disruption ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: This communication describes the development of genetic tools for the yeast Yamadazyma ohmeri. Nystatin enrichment proved highly effective for isolating various auxotrophic strains, which were classified by complementation analysis. Biosynthetic genes encoding known biochemical functions were isolated by polymerase chain reaction, including YoLEU2 and YoURA3 that were sequenced. Using these homologous genes as selective markers, DNA transformation was accomplished by electroporation. Transformation with pBR322-based plasmids, cut within the coding region of the homologous marker gene, yielded 20 to 50 stable transformants per μg of DNA. In about 80% of the cases, integration of plasmid DNA sequence occurred by homologous recombination of a single plasmid into the chromosome. Excision of the plasmid permitted gene replacement, as illustrated by the substitution of a wild-type URA3 gene by an in vitro generated deletion.Sequences conferring extrachromosomal replication were isolated from Y. ohmeri DNA. Plasmids based on pBR322 carrying such an ARS and either selective markers transformed at 104/μg and were shown to replicate freely in Y. ohmeri at an approximate copy number of 40. Unexpectedly, we observed that BS-SKR derivatives carrying either YoLEU2 or YoURA3 but no Y. ohmeri ARS also replicated extrachromosomally. Linearization of transforming plasmids within regions homologous or not to chromosomal sequences stimulated transformation frequencies up to four-fold. The sequences are available for consultation under EMBL accession number Z35101 for YoLEU2 and Z35100 for YOURA3.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320101209
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