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  • 1985-1989  (2)
  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 205 (1986), S. 146-152 
    ISSN: 1617-4623
    Keywords: Cellobiase ; Oligodeoxyribonucleotide ; Expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The β-glucosidase from ATCC 21400, anAgrobacterium species, was purified to homogeneity. The protein was cleaved with cyanogen bromide and the peptides were purified by reversed phase FPLC. The partial amino acid sequence for one peptide was determined by automated Edman degradation. The sequence was used to synthesize a mixture of oligodeoxyribonucleotides which was used as a hybridization probe to identify a recombinant DNA clone carrying the gene for β-glucosidase. A single clone was isolated which expressed an enzymatic activity that hydrolyzed several β-glucosides. The enzymatic activity produced by this clone could be adsorbed by rabbit antiserum raised against theAgrobacterium enzyme. The direction of transcription of the β-glucosidase gene was determined by verifying the DNA sequence 3′ to the oligodeoxyribonucleotide probe binding site. After subcloning the gene a high level of expression was obtained in the plasmid vector pUC18 using the lacZ gene promoter.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Archives of microbiology 152 (1989), S. 572-577 
    ISSN: 1432-072X
    Keywords: Chlamydomonas reinhardtii ; Gametic differentiation ; Light requirement ; Light-pulse-studies
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Gametes of Chlamydomonas reinhardtii synthesize numerous proteins not observed in vegetative cells and vice versa. Gametogenesis induced changes in gene expression were confirmed by SDS-PAGE of in vitro translation products using total RNA from gametes and vegetative cells. Vegetative cells and gametes thus represent two cell types with distinct patterns of gene expression. The generation of mature gametes from liquid cultures of asynchronously growing vegetative cells was dependent on light. This light requirement could not be substituted for by an organic source of energy and carbon, indicating that light serves as a signal in gametogenesis. The light signal was shown to become effective only after preincubation in nitrogen-free medium. This delayed competence for light indicates that the two external signals — nitrogen-starvation and light —may function in sequence. Execution of the light dependent step in gamete formation required cytoplasmic protein synthesis and RNA synthesis.
    Type of Medium: Electronic Resource
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