林可霉素转运蛋白基因lmrC的功能分析

主管：中华人民共和国教育部
主办：上海交通大学
ISSN 1006-2467 CN 31-1466/U

上海交通大学学报（自然版） ›› 2014, Vol. 48 ›› Issue (02): 159-163.

• 生物科学 • 下一篇

林可霉素转运蛋白基因lmrC的功能分析

徐晶晶1，吴杭1，孟思童2，刘瑞华3，黄训端1，张部昌1，白林泉2

(1.安徽大学生命科学学院，合肥 230601； 2.上海交通大学微生物代谢国家重点实验室，上海 200240；
3.安徽皖北药业股份有限公司，安徽宿州 234000)

收稿日期:2013-01-12 出版日期:2014-02-28 发布日期:2014-02-28
基金资助:
国家自然科学基金（31300081），安徽大学博士科研启动经费，微生物代谢国家重点实验室开放课题（MMLKF1305）资助项目

Functional Analysis of Lincomycin Transporter Gene lmrC in Streptomyces Lincolnensis LC-G

XU Jingjing1,WU Hang1,MENG Sitong2,LIU Ruihua3 HUANG Xunduan1,ZHANG Buchang1,BAI Linquan2

(1.School of Life Science, Anhui University, Hefei 230601, China; 2.State Key Laboratory of Microbial Metabolism, Shanghai Jiaotong University, Shanghai 200240, China; 3.Anhui Wanbei Pharmaceutical Co. Ltd., Suzhou 234000, Anhui, China)

Received:2013-01-12 Online:2014-02-28 Published:2014-02-28

摘要/Abstract

摘要：

为了研究腺苷三磷酸结合盒转运蛋白LmrC是否参与林可霉素的生物合成，通过同源重组在林可霉素高产菌株LC-G中构建了lmrC的缺失突变株XJJ7.高效液相色谱(HPLC)分析和抗性检测显示，相比出发菌株LC-G，XJJ7的林可霉素产量下降了55%，而且耐受林可霉素的最高质量浓度降低了50%.进一步实时荧光定量聚合酶链反应(PCR)分析发现，突变株XJJ7中林可霉素生物合成基因lmbA和lmbR的转录明显低于LC-G，但调控基因lmbU的转录不变.在LC-G中lmrC的过表达不能进一步提高产量，也不影响lmbA、lmbR和lmbU基因的转录，但过表达菌株对林可霉素的抗性显著增强.结果表明，lmrC可以通过影响生物合成基因的转录来调节林可霉素的产生，并赋予产生菌一定的抗性.

关键词: 林可链霉菌, 林可霉素, 腺苷三磷酸结合盒转运蛋白, 抗性

Abstract:

To investigate the role of lincomycin transporter LmrC in lincomycin biosynthesis, lmrC was deleted in a lincomycin high-yielding strain LC-G by homologous recombination. HPLC analysis and resistance assay indicated that lincomycin production and self-resistance in XJJ7, and the lmrC mutant were decreased by 55% and 50%, respectively. Quantitative real-time polymerase chain reaction(PCR) analysis showed that the transcriptional levels of structure genes lmbA and lmbR were drastically reduced, whereas the expression of a regulatory gene lmbU did not change in XJJ7. Moreover, overexpression of lmrC in LCG could not increase the lincomycin yield and the expression of lmbA, lmbR and lmbU as well. However, the resistance to lincomycin was improved in the lmrCoverexpressed strain. Therefore, lmrC was proved to be involved in lincomycin biosynthesis by affecting the transcription of structure genes and confer lincomycin resistance to Streptomyces lincolnensis.

Key words: Streptomyces lincolnensis, lincomycin, ATP-binding cassette (ABC) transporter, resistance

中图分类号:

Q 93

徐晶晶1，吴杭1，孟思童2，刘瑞华3，黄训端1，张部昌1，白林泉2. 林可霉素转运蛋白基因lmrC的功能分析[J]. 上海交通大学学报（自然版）, 2014, 48(02): 159-163.

XU Jingjing1,WU Hang1,MENG Sitong2,LIU Ruihua3 HUANG Xunduan1,ZHANG Buchang1,BAI Linquan2. Functional Analysis of Lincomycin Transporter Gene lmrC in Streptomyces Lincolnensis LC-G[J]. Journal of Shanghai Jiaotong University, 2014, 48(02): 159-163.

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