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目的:建立抗生素菌株选育的新方法。方法:变铅青链霉菌和天蓝色链霉菌A3(2)的relA突变株的合成微量放线紫红素(Act)和十一烷基灵菌红素(Red),通过诱导对卷曲霉素的抗性,筛选能够恢复或活化Act和Red合成的突变株,并应用蛋白质印迹分析考察活化或恢复Act或Red的合成与表达两个抗生素生物合成途径中的调节蛋白ActII-ORF4或RedD之间关系。结果:一些卷曲霉素抗性突变株(cap)的突变能够恢复relA突变株被损伤的抗生素合成和活化变铅青链霉菌的抗生素(Act和Red)合成,但没有恢复ppGpp合成,暗示着cap突变能够使天蓝色链霉菌不依赖ppGpp分子启动抗生素的合成;变铅青链霉菌66的cap突变株展现不同的表型,其基因突变的位点或类型可能互不相同。结论:建立了一个新的抗生素生产菌株选育方法,即诱导生产菌株对卷曲霉素的抗性。
Objective: To establish a new method of antibiotic strain selection. Methods: The synthetic actinomycetes (Act) and undecylmethionine (Red) of relA mutants of Streptomyces lividans and Streptomyces coelicolor A3 (2) Resistant and screening for mutants capable of restoring or activating Act and Red synthesis and applying Western blot analysis to examine the relationship between the activation or restoration of Act or Red synthesis and the expression of the regulatory proteins ActII-ORF4 or RedD in the two antibiotic biosynthetic pathways . Results: Some capreotenic cap mutations have been shown to restore the synthesis of the relA mutant impaired antibiotic and to activate the synthesis of the antibiotic Streptomyces lividans (Act and Red), but did not restore ppGpp synthesis, suggesting that cap The mutation enables Streptomyces coelicolor not to rely on the initiation of antibiotic synthesis by the ppGpp molecule. Cap mutants of Streptomyces lividans 66 exhibit different phenotypes, and their loci or types of mutations may differ from each other. Conclusion: A new breeding method of antibiotic producing strains was established, which is to induce the producing strains to be resistant to capreomycin.