庆大霉素生物合成基因簇中抗性基因gmrB及gmrA的功能研究.PDF

. 688 . 中国抗生素杂志2018年6月第43卷第6期 文章编号：1001-8689(2018)06-0688-08 遗传育种与生物合成 庆大霉素生物合成基因簇中抗性基因gmrB 及gmrA 的功能研究 * 万云凤 连榕 洪文荣 石贤爱 (福州大学，生物科学与工程学院，福州350116) 摘要：目的 研究绛红色小单孢菌(Micromonospora purpurea)G1008中抗性基因gmrB 与gmrA功能，为阐明庆大霉素生物合 成机制奠定基础。方法 以红霉素抗性基因ermE分别置换gmrB和gmrA基因，构建突变株GerB 102和GerA 102 。借助TLC和MS分 析突变株代谢产物是否变化，同时检测突变株耐受自身代谢产物的能力是否发生变化。结果 突变株GerB 102代谢产物没有发 生明显变化，仍主要积累庆大霉素C化合物，而GerA 102代谢产物发生变化。庆大霉素耐受性试验结果显示，GerB 102和G 1008 耐庆大霉素浓度达到6000U/mL 以上，而GerA 102不足50U/mL 。结论 gmrB 既非庆大霉素生物合成的关键基因，也非关键的抗 性基因，而gmrA是庆大霉素抗性的关键基因。 关键词：gmrB基因；gmrA基因；庆大霉素；基因表达；基因工程 中图分类号：Q933 文献标志码：A Study on the function of resistant genes gmrB and gmrA in gentamicin biosynthetic gene cluster Wan Yun-feng, Lian Rong, Hong Wen-rong and Shi Xian-ai (College of Biological Science and Technology, Fuzhou University, Fuzhou 350116) Abstract Objective Characterizing the functions of gmrB and gmrA of Micromonospora purpurea G1008 to elucidate the gentamicin biosynthetic mechanisms. Methods The mutant strains GerB102 and GerA102 were constructed by replacing of gmrB and gmrA with the erythromycin resistance gene ermE, respectively. Then the possible alterations of the metabolites in these mutants were analyzed by TLC and MS. The resistance of the mutants was also detected against their own metabolites (gentamicin), respectively. Results The metabolite profile of GerB102 was not significantly changed where gentamicin C components still accumulated, while the metabolite profile of GerA102 was changed. Similar to the native G1008 strain, the tolerance to gentam