抗感染海洋天然产物marformycin生物合成基因簇中调控基因及转运基因的功能研究  

Functional investigation of the regulatory and transportation genes in the biosynthetic gene cluster of marine-derived anti-infection marformycins

作　　者：李宏志[1,2] 刘静[1] 李青连[1] 鞠建华[1] 

机构地区：[1]中国科学院热带海洋生物资源与生态重点实验室,广东省海洋药物重点实验室,中国科学院南海海洋研究所,广东广州510301 [2]中国科学院大学,北京100049

出　　处：《中国海洋药物》2016年第1期10-18,共9页Chinese Journal of Marine Drugs

基　　金：国家自然科学基金面上项目(41306146,31290233,81425022)资助

摘　　要：目的对海洋放线菌Streptomyces drozdowiczii SCSIO 10141抗感染次级代谢产物marformycins生物合成基因簇中的调控基因mfnM与转运基因mfnR进行生物信息学分析和功能鉴定。方法利用生物信息学对mfnM与mfnR的功能进行预测,利用PCR-targeting技术对mfnM与mfnR进行体内阻断,构建双交换突变株△mfnM和△mfnR,对突变株进行发酵并利用HPLC对发酵液进行分析。结果突变株△mfnM和△mfnR丧失了生产marformycins代谢产物的能力。结论初步阐明了mfnM与mfnR的功能,mfnM编码1个正调控因子,对marformycins的生物合成起正调控作用;而mfnR编码的ABC转运蛋白,负责marformycins的胞外转运。Objective To identify and characterize the regulatory and transportation genes in anti-infection marformycins biosynthetic gene cluster in Streptomyces drozdowiczii SCSIO 10141.Methods The functions of mfnM and mfnR were first predicted by bioinformatics analysis.Then mfnM and mfnR were inactivated using PCR-targeting mutagenesis methodology to generate the double cross mutant strains△mfnM and △mfnR,respectively.The mutant strains were fermented and the fermentation broths were analyzed by high performance liquid chromatography（HPLC）.Results The production of marformycins was abolished in the△mfnMand△mfnR mutants.Conclusion mfnMencoded a positive regulator involved in the regulation of marformycin biosynthesis,and mfnRencoded an ABC-transporter involved in transportation of maformycins.

关 键 词：STREPTOMYCES drozdowiczii SCSIO 10141 marformycin 调控基因 抗性基因 ABC转运蛋白 

分 类 号：R915[医药卫生—微生物与生化药学]