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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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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