orfX基因整合表达在阿维链霉菌工业育种中的应用  

Application of Integrative Expression of orfX Gene in the Industrial Breeding of Streptomyces avermitilis

作　　者：范与庆[1] 闵勇[1] 熊伟[2] 陈芬[1] 梁运祥[1] 吕和平[3] 邢仁昌[3] 郑应华[1] 

机构地区：[1]华中农业大学农业微生物国家重点实验室,武汉430070 [2]清华大学生物科学与技术系,北京100084 [3]北京生物医药研究所,北京100091

出　　处：《农业生物技术学报》2007年第6期1048-1052,共5页Journal of Agricultural Biotechnology

摘　　要：阿维链霉菌(Streptomyces avermitilis)基因组中含有orfX基因,通过增加orfX基因剂量可以提高阿维菌素的产量。研究以pIJ8600为出发质粒,构建了含红霉素抗性基因启动子(ermEp*)和orfX结构基因DNA片段的重组质粒pBIB827。通过接合转移将pBIB827导入阿维菌素的工业菌株BIB9903中,得到稳定高产的重组菌株BIB0827。高效液相(HPLC)分析结果表明,与出发菌株BIB9903相比,BIB0827发酵产物阿维菌素产量提高了1.5倍,达到3350μg/mL。传代稳定性实验证明,BIB0827传代10代后依然稳定,在工业生产上有较大的应用价值。The level of avermectin produced by Streptomyces avermitilis could be stimulated by increasing the dosage of orfX gene. Recombinant plasmid pBIB827,which contained ermEp＊ and structural gene of orfX, was derived from pU8600. The recombinant strain BIB0827 was obtained by introducing pBIB827 into the industrial strain BIB9903 through intergeneric conjugal transfer. HPLC assays showed the level of avermectin produced by BIB0827 reached 3 350 μg/mL, 1.5-fold increasing when compared with BIB9903. The characteristic of high-level producing strain BIB0827 was stable after ten rounds of non-selective growth, which can be applied in the industrial production of avermectin.

关 键 词：orfX基因 接合转移 整合表达 阿维链霉菌 阿维菌素 

分 类 号：S188[农业科学—农业基础科学]