摘要
近年的细菌耐药性监测结果发现,肺炎克雷伯菌对庆大霉素等氨基糖苷类抗生素的耐药率已达40%左右。耐药的主要机制为细菌产生N-乙酰转移酶AAC(3)-V,使其抗菌活性不能发挥。不同菌株产生的钝化酶及其编码基因有一定的差异。作者从华山医院分离的肺炎克雷伯菌耐药株,用PCR方法从该菌株的一分子量约85×10^3的接合型质粒上扩增出全长基因,连接pUC18。
For the purpose of crystallography and kinetic study of N-acetyltranferase (3)-V , the encoding gene , aac (3)-Va was cloned from K.pneumonia and expressed in E. coli. The aac (3) -Va amplified from a conjugate plasmid of 85Md by PCR was ligated in pUC 18. From the transformants ,we isolated the clone encoding N-acetyltransferase (3)-V. It was proved by restriction fragments analysis and sequencing, more than 95% sequence of this gene was identical to that of S. marcescens, Most of the amino acids substitution located in C terminal. This gene ligated with pLY4 was successfully expressed in E. coli JF1125 and DH5a.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
1996年第6期409-412,共4页
Chinese Journal of Microbiology and Immunology
基金
国家自然科学基金