圈卷产色链霉菌尼可霉素生物合成基因──sanJ的克隆及功能研究  

Cloning and Function of sanJ──a Gene Involved in Nikkomycins Biosynthesis of Streptomyces ansochromogenes

作　　者：聂丽平[1] 田宇清[1] 谭华荣[1] 

机构地区：[1]中国科学院微生物研究所

出　　处：《Acta Genetica Sinica》2001年第2期188-195,共8页

基　　金：国家自然科学基金!(No>39830010);国家杰出青年基金项目(No.39925002)&&

摘　　要：以尼可霉素生物合成相关的基因片段为探针，从圈卷产色链霉菌cosmid基因文库中筛选到1个大约7.5kb的DNA片 段，将此DNA片段克隆到载体pBluescripM13-的Kpn I位点，得到了重组质粒pNL2200。对pNL2200中外源DNA片段进行了一系列的亚克隆及部分核苷酸序列分析，结果表明，2.3kb的Sal I-BamH DNA片段中含有1个完整的开放阅读框，起始密码子为271位的GTG，终止密码子为1954位的TGA，该基因的大小为1 686bp，编码1个大小为561个氨基酸的蛋白质产物。利用blastx程序在蛋白质数据库中进行同源比较，结果揭示此基因产物与腺苷酸形成酶超家族的连接酶有44%的一致性。此外，该基因的破坏导致圈卷产色链霉菌尼可霉素生物合成能力的丧失，证明它是尼可霉素生物合成所必需的，命名其为sanJ。A 7.5kb DNA fragment was identified from the cosmid library of Streptomyces ansochromogenesus using a partial DNA fragment involved in the nikkomycin biosynthesis as a probe. It was cloned into pBluescript M13-and the resulting recombinant plasmid was designated as pNL2200. The nucleotide sequence of 2.3kb Sal I-BamH DNA fragment was determined and analysed. The result indicated that the fragment contains one complete open reading frame (ORF); The start codon is GTG at 271 position and the stop codon is TGA at 1 954 position. It contains 1686 base pairs and encodes a protein with 561amino acids. The deduced product has 44% identity with the ligase belong to the superfamily of adenylate-formingenzyme. The experiment of gene disruption showed that the sanJ gene was closely relatedtonikkomycins biosynthesis of Streptomyces ansochromogenes. It was designated as sanJ.

关 键 词：圈卷产色链霉菌 尼可霉素 生命合成 腺苷酸形成酶 基因 克隆 

分 类 号：Q785[生物学—分子生物学]