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目的将糖多孢红霉菌bldD基因转入活跃链霉菌(Streptomyces actuosus)N-H,探讨其对诺西肽产量的影响。方法将bldD基因连接到链霉菌整合型表达载体pZMW上构建pZMW-bldD质粒,利用PEG介导的原生质体转化方法将重组质粒pZMW-bldD及对照质粒pZMW-vgb分别导入活跃链霉菌,通过对枯草芽孢杆菌抑菌试验和分光光度计法测定发酵液中诺西肽产量。结果成功构建了活跃链霉菌N-H/pZMW-bldD工程菌株及对照菌株N-H/pZMW-vgb。与出发菌株N-H相比,N-H/pZMW-bldD菌株诺西肽产量提高了68%;与对照菌株N-H/pZMW-vgb相比,N-H/pZMW-bldD菌株诺西肽产量提高了19%。结论在活跃链霉菌中导入糖多孢红霉菌bldD基因可以提高诺西肽产量,其作用效果较vgb基因更为明显,说明bldD基因对抗生素产量的提高具有广谱性。
Objective To transfer the bldD gene from Streptomyces actuosus to N-H and investigate its effect on the yield of nosiheptide. Methods The bldD gene was ligated into Streptomyces integrative expression vector pZMW to construct pZMW-bldD plasmid. The recombinant plasmid pZMW-bldD and the control plasmid pZMW-vgb were respectively introduced into Streptomyces fulgidus by PEG-mediated protoplast transformation. Bacillus subtilis bacteriostatic test and spectrophotometer method for determination of nosiheptide in fermentation broth. Results The constructed Streptomyces N-H / pZMW-bldD engineering strain and the control strain N-H / pZMW-vgb were successfully constructed. Compared with the starting strain N-H, the yield of nosin produced by N-H / pZMW-bldD strain was increased by 68%. Compared with the control strain N-H / pZMW-vgb, the yield of nosiheptide by N-H / pZMW-bldD strain increased by 19%. Conclusion The introduction of the bldD gene into Streptomyces can increase the production of nosiheptide and its effect is more obvious than that of vgb gene, indicating that bldD gene has a broad spectrum of antibiotic production.