| 摘要 |
PURPOSE: Provided are base sequence and amino acid sequence of enzyme used for synthesizing spectinomycin. In particular, provided is gene sequencing of methyltransferase and glycosyltransferase originated from Streptomyces spectabilis ATCC 27741 for synthesizing antibiotic, spectinomycin. CONSTITUTION: A base sequence and amino acid sequence of methyltransferase and glycosyltransferase are determined by following steps of; (a)performing PCR amplification of gene library using genome of S. spectabilis ATCC 27741 and isolating clone having two cosmids pHCG121 and pHCG171; (b)transforming Streptomyces lividans, which is sensitive to spectinomycin, with two cosmids and examining two cosmids having an effect on biosynthesis of spectinomycin by identifying resistance of transformant against spectinomycin; (c)subcloning all the BamHI fragments of two cosmids into pBluescript KS(+) and identifying both directions of base sequence, thereby methyltransferase gene is identified in recombinant plasmid pHCG1714 and also, area coding spcG and spcM is identified in BamHI inserting fragment of the recombinant plasmid pHCG1714; (e)sequencing both spcG and spcM; and (f)identifying amino acid homology between SpcG and glycosyltransferase and amino acid homology between SpcM and methyltransferase.
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