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  • 1
    Electronic Resource
    Electronic Resource
    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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