Promotion of markerless deletion of the actinorhodin biosynthetic gene cluster in Streptomyces coelicolor  

作　　者：Zhiqun Lu Pengfei Xie Zhongjun Qin 

机构地区：[1]Key Laboratory of Synthetic Biology, Shanghai Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, TheChinese Academy of Sciences, Shanghai 200032, China

出　　处：《Acta Biochimica et Biophysica Sinica》2010年第10期717-721,共5页生物化学与生物物理学报（英文版）

基　　金：This work was supported by grants from National Natural Science Foundation of China （30770045 and 30870067）, National ＇863＇ project （2007AA021503） and the Chinese Academy of Sciences project （KSCX2-YW-G-069） to Z.Q.

摘　　要：The standard gene disruption and replacement to delete the actinorhodin biosynthetic gene cluster （Act） in Streptomyces coelicolor was inefficient, and the polymerase chain reaction-targeting of the cosmid could efficiently delete the Act, but still was a time-consuming procedure for markerless gene replacement. By using optimal Streptomyces codons, we synthesized a sceS gene encoding identical amino acid sequence as the chromosome rarecutting meganuclease I-sce I of the Saccharomyces cerevisiae mitochondria. Expression of sceS gene in S. coelicolor resulted in promotion of homologous recombination and subsequently, successful achieved markerless deletion of the Act. The sceS system may be useful for the sequential markerless deletions of chromosomal segments in Streptomyces.在 Streptomyces coelicolor 的基因混乱和代替聚类删除 actinorhodin biosynthetic 基因的标准(行为) 是低效的，并且 cosmid 指向反应的聚合酶链能高效地删除行为，但是仍然是为无标记的基因代替的一个费时间的过程。由使用最佳的 Streptomyces codons，我们综合了作为染色体编码相同氨基酸顺序的 sceS 基因稀罕切的 meganuclease I-sce 我 Saccharomyces cerevisiae 线粒体。在 S 的 sceS 基因的表示。coelicolor 导致了相应再结合并且随后的提升，行为的成功的完成的无标记的删除。sceS 系统可能为在 Streptomyces 的 chromosomal 片断的顺序的无标记的删除是有用的。

关 键 词：STREPTOMYCES sceS gene antibioticbiosynthetic gene gene replacement 

分 类 号：Q939.117[生物学—微生物学] TQ920.1[轻工技术与工程—发酵工程]