Objective To construct the mutant of Saccharopolyspora erythraea in which SACE_0065, SACE_2559 or SACE_2565 gene is knocked out respectively, and to make preliminary function analysis of those genes. Methods Linear DNA containing thiostrepton resistance gene ( tsr ) with upper and down homologous fragments of the target inactivated gene around tsr was transformed into Sac. erythraea A226 under PEG mediation. By chromosomic homologous recombination, the mutant in which the target gene was disrupted could be screened out with thiostrepton choice. The mutant strains were com-pared with the original strain A226 and the △bldD mutant in differentiation of aerial hyphae into spores. Results and Con-clusion The growth of aerial hyphae and the differentiation of spores of △SACE_0065 and ASACE_2559 mutants were delayed, while△SACE_2565 mutant did not change obviously compared with the original strain A226. SACE_0065 and SACE_2559 in Sac. erythraea may be involved in regulating the morphological differentiation, and could be regulatory genes in the development of spores.%目的 构建红色糖多孢菌ΔSACE_0065、ΔSACE_2559和ΔSACE_2565三个突变体并研究其功能.方法 将待失活目的 基因的上下游同源片段依次连接到pUCTSR质粒Thio抗性基因(tsr)两侧,利用PEG介导的原生质体转化法将线性DNA同源片段转入红色糖多孢菌A226内,通过同源重组使目的 基因失活,最后将构建出的三个突变株与出发菌株A226及ΔbldD突变株进行比较,观察孢子生长有无差异.结果 与结论 成功构建ΔSACE_0065、ΔSACE_2559和ΔSACE_2565三个突变株;与A226相比,ΔSACE_0065和ΔSACE_2559气生菌丝和孢子形成推迟,而ΔSACE_2565突变株的生长情况则无明显变化,提示红色糖多孢菌SACE_0065、SACE_2559基因参与其形态分化的调控.
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