阿扎霉素F产生菌链霉菌211726基因转移系统的建立

摘要

The objective of this study is to establish the gene transfer system of strain Streptomyces sp. 211726 producing azalomycin F,which can be used for genetic manipulations such as gene knock-out and expression of foreign genes. Intergeneric genetic transfer system of Streptomyces sp. 211726 producing azalomycin F was constructed by conjugating integrative plasmid pSET152 with pIB139. Results showed that 25 mg/mL apramycin may be used to efficiently screen conjugants. PCR verification revealed that exogenous plasmid was successfully integrated in the chromosomal DNA of Streptomyces sp. 211726. The continuous passage culture experiment demonstrated that transformed pSET152 and pIB139 of conjugants were stably inherited.%旨在建立阿扎霉素F产生菌链霉菌211726的基因转移系统，以便基因敲除和外源基因表达等遗传操作。以整合型质粒pSET152和pIB139为出发质粒，通过接合转移构建了阿扎霉素F产生菌链霉菌211726的基因转移系统。结果显示25μg/mL阿泊拉霉素可有效筛选接合子。经PCR验证，质粒成功整合到菌株链霉菌211726基因组中，接合子经多次传代后，导入的质粒pSET152和pIB139仍稳定整合于接合子基因组上。