摘要
利用DNA同源重组原理敲除S.lincolnensisNRRL 2936中lmbU基因,筛选获得该基因缺陷株S.lincolnensisJLUa2,其不具备林可霉素生物合成能力。利用构建的不同重组质粒转化缺陷株,发现含有质粒pHBUYX335的缺陷株重新获得了林可霉素生物合成能力。单一回补下游基因lmbY和lmbX不能使缺陷株重新恢复林可霉素合成能力;而同时回补lmbU基因和下游基因lmbY和lmbX却能回补这种缺陷,缺陷株林可霉素合成能力的丧失不仅是由下游基因的破坏而引起,证实了lmbU基因是林可霉素生物合成的关键基因。根据不同重组质粒上装载的基因片段性质推测了lmbU基因可能的启动子。
Lincomycin nonproducing mutant of S. lincolnensis NRRL 2936, named as S. lincolnensis JLUa2, was generated by gene knockout of lrnbU with homologous recombination. Different recombinant plasmids were transformed into the mutant. The mutant harboring the recombinant plasmid pHBUYX335 showed a refreshing ability of producing lincomycin. The recovery of biosynthetic ability can be achieved by retro-complementation of not only two downstream genes lmbY and lmbX, but also lmbU and these downstream genes, which demonstrates that lmbU plays a key role in the procedure of lincomycin biosynthesis. Analysis of gene fragments cloned to different recombinant plasmids, and its probable promoter is deduced.
出处
《华东理工大学学报（自然科学版）》
EI
CAS
CSCD
北大核心
2008年第1期60-65,共6页
Journal of East China University of Science and Technology
关键词
同源重组
敲除
遗传回补
林可霉素生物合成
homologous
knockout
retro-complementation
lincomycin biosynthesis
