摘要
目的:建立工业生产菌株阿维链霉菌QL0827的接合转移体系。方法:优化接合转移条件,建立该菌株的接合转移体系;应用优化后的接合转移体系,将基因中断质粒导入该菌,定向敲除基因组上的序列;再将携带基因组上特定序列的整合型质粒和复制型质粒导入该菌体,建立该菌株的基因过表达体系。结果:相比标准的接合转移方法,优化后接合转移效率提高了近150倍,达到2.45×10-6;并成功地将pks3基因簇的部分序列删除。结论:建立了阿维链霉菌QL0827的接合转移体系。
Objective： To develop a conjugal transfer system for the industrial strain Streptomyces avermitilis QL0827. Methods： Firstly, optimize conjugal transfer conditions for the establishment of the strain conjugal trans-fer system. Then, apply the optimized conjugal transfer system to intoduce the gene-disruption plasmids into thespores for knocking out the target sequence in the genome and the integrative or replicative plasmids carrying spe-cific genes into the spores to establish the gene-overexpressing system in the strain. Results： By using the opti-mized method, the conjugal transfer efficiency has been improved about 150 times compared with the standardmethod and up to 2.45×10^（-6）. And the partial sequence of pks3 cluster has been deleted. Conclusion： Conjugaltransfer system between E.coli and the industrial strain S.avermitilis QL0827 has been achieved.
出处
《生物技术通讯》
CAS
2016年第2期188-191,共4页
Letters in Biotechnology
关键词
阿维链霉菌
接合转移
基因敲除
Streptomyces avermitilis
conjugal transfer
gene knock-out
