李宏志,刘静,李青连,等.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[J].中国海洋药物,2016,35(1):10-18.
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
Fuctional investigation of the regulatory and transportation genes in the biosynthetic pathway of marine-derived anti-infective marformycins
投稿时间:2015-04-10  修订日期:2015-04-26
DOI:
中文关键词:  Streptomyces drozdowiczii SCSIO 10141  marformycin  调控基因  抗性基因  ABC转运蛋白
English Keywords:Streptomyces drozdowiczii SCSIO 10141  marformycin  regulatory gene  resistant gene  ABC-transporter
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作者单位E-mail
李宏志 中国科学院南海海洋研究所 lnbxlhz@163.com 
刘静 中国科学院热带海洋生物资源与生态重点实验室  
李青连 中国科学院热带海洋生物资源与生态重点实验室  
鞠建华* 中国科学院热带海洋生物资源与生态重点实验室
中国科学院大学 
jju@scsio.ac.cn 
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中文摘要:
      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编码一个正调控因子,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。
English Summary:
      Objective To identify and characterize the regulatory and transportation genes in anti-infective marformycins biosynthetic pathway 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 △mfnM and △mfnR mutants. Conclusion mfnM encodes a positive regulator involved in the regulation of marformycin biosynthesis, and mfnR encodes an ABC-transporter involved in transportation of maformycins.
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