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目的:通过考察庆大霉素抗性突变株,发现庆大霉素抗性与加强抗生素合成能力之间关系。方法:通过诱导对庆大霉素的低水平抗性,筛选能够加强抗生素合成的突变株;通过在没加抗生素的琼脂平板系列传代,优化突变株的稳定性,并应用蛋白质印迹分析考察加强抗生素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:大约3%-20%的庆大霉素低水平抗性突变株(gen)能够加强抗生素的合成水平。在变铅青链霉菌的庆大抗性突变株中,活化Act和Red的合成与过量表达两个抗生素生物合成途径中的调节蛋白ActⅡ-ORF4或RedD之间密切相关。结论:本研究建立一个新的抗生素生产菌株选育方法,即诱导链霉菌和细菌对庆大霉素的低水平抗性。
OBJECTIVE: To investigate the relationship between gentamycin resistance and antibiotic biosynthesis through investigating gentamicin resistant mutants. Methods: By inducing the low level resistance to gentamicin, we screened the mutants that could enhance the antibiotic synthesis. The stability of the mutants was optimized by passage on the agar plate without antibiotics, and the antibiotic The relationship between the synthesis and expression of Act II-ORF4 or RedD in the two antibiotic biosynthetic pathways was demonstrated. Results: Approximately 3% -20% of gentamicin low-level resistant mutants (gen) potentiate antibiotic synthesis. Among the mutagenized mutants of Streptomyces lividans, the activation of Act and Red is closely related to the overexpression of the regulatory protein ActII-ORF4 or RedD in the two antibiotic biosynthetic pathways. Conclusion: This study established a new breeding method of antibiotic production strain, that is, induced the low level of gentamicin resistance of Streptomyces and bacteria.