产黄青霉转化载体pPIPKA的构建及青霉素工业生产菌株的转化研究  

CONSTRUCTION OF VECTOR pPIPKA AND TRANSFORMATION OF PENICILLIUM CHRYSOGENUM INDUSTRIAL STRAIN

作　　者：王富强[1] 任志红[1] 徐平[1] 赵颖[1] 修建新[1] 朱研研 贺秉坤[1] 

机构地区：[1]华北制药集团新药研究开发有限责任公司

出　　处：《菌物学报》2004年第1期66-72,共7页Mycosystema

基　　金：国家科技攻关计划(2002BA711A16);国家技术创新项目计划资金资助

摘　　要：以腐草霉素(phleomycin)抗性为选择标记,构建了用于青霉素生产菌—产黄青霉Penicillium chrysogenum的转化载体pPIPKA。 以此转化当前的青霉素生产菌株NCPC-10086,建立了高效的工业生产菌株的基因转化体系,转化效率最高达18个转化子/g DNA。对转化子进行了PCR及Southern杂交分析,结果表明,外源基因整合到了宿主的染色体上。Filamentous fungus vector, pPIPKA, was constructed and used to establish a high efficiency gene transfer system for present penicillin industrial strain NCPC-10086. In pPIPKA, phleomycin resistance gene (Sh ble) fused to Pipns, a fungal promoter derived from Penicillium chrysogenum isopenicillin N synthetase gene, was used as a dominant selection marker. The frequency of transformation with pPIPKA was up to 18 transformants per g DNA. Analysis of the transformants by PCR and Southern blot showed the foreign DNA had been integrated into the host抯 chromosome.

关 键 词：基因转化 显性选择标记 腐草霉素 青霉素 

分 类 号：TQ465[化学工程—制药化工]