Site-directed Mutagenesis of Streptomyces avermitilis aveD Gene

作     者：汤晖 张利平 

作者机构：河北大学生命科学学院河北省微生物多样性研究与应用实验室河北保定071002 

出 版 物：《Agricultural Science & Technology》 (农业科学与技术（英文版）)

年 卷 期：2011年第12卷第10期

页      面：1424-1426页

学科分类：090403[农学-农药学(可授农学、理学学位）] 09[农学] 0904[农学-植物保护] 

基　　金：Supported by Special Fund for Doctors in Universities in Hebei Province(B2003201) 

主　　题：PCR-driven overlap extension aveD gene Site-directed mutagenesis 

摘      要：[Objective] The aim of this study was to produce Streptomyces avermitilis strain with site-directed mutagenesis in aveD gene,and so as to provide theoretical basis for genetic breeding of ***.[Method] PCR-driven overlap extension was conducted for the site-directed mutagenesis in aveD gene;the mutated aveD gene then was used to construct vector pDC3（pKC1139∷aveD） via molecular manipulations like in vitro enzyme digestion and ligation;the vector pDC3（pKC1139∷aveD） was then introduced to aveD deletion mutant 489 of avermectin-producing strain *** 76-9.[Result] Mutant strain 536 of site-directed mutagenesis of *** 76-9 was obtained by homologous *** sequencing results show that the sixty-ninth base C in aveD-coding region of mutant 536 was substituted by T,and the corresponding amino acid Thr was mutated to be Ile.[Conclusion] This study laid basis for the development of strains specifically producing avermectin B.