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orfX基因整合表达在阿维链霉菌工业育种中的应用
引用本文:范与庆,闵勇,熊伟,陈芬,梁运祥,吕和平,邢仁昌,郑应华.orfX基因整合表达在阿维链霉菌工业育种中的应用[J].农业生物技术学报,2007,15(6):1048-1052.
作者姓名:范与庆  闵勇  熊伟  陈芬  梁运祥  吕和平  邢仁昌  郑应华
作者单位: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
4. 华中农业大学农业微生物国家重点实验室,武汉,430070;北京生物医药研究所,北京,100091
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。本研究以pIJ8600为出发质粒,构建了含红霉素启动子(ermEp*)和orfX结构基因DNA片段的整合型表达载体pWJ827。通过接合转移将pWJ827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。

关 键 词:orfX基因  接合转移  整合表达  阿维链霉菌  阿维菌素
文章编号:1006-1304(2007)06-1048-05
收稿时间:2007-03-27
修稿时间:2007-05-06

Application of Integrative Expression of orfX Gene in the Industrial Breeding of Streptomyces avermitilis
FAN Yu-qing,MIN Yong,XIONG Wei,CHEN Fen,LIANG Yun-xiang,LV He-ping,XING Ren-chang,ZHENG Ying-hua.Application of Integrative Expression of orfX Gene in the Industrial Breeding of Streptomyces avermitilis[J].Journal of Agricultural Biotechnology,2007,15(6):1048-1052.
Authors:FAN Yu-qing  MIN Yong  XIONG Wei  CHEN Fen  LIANG Yun-xiang  LV He-ping  XING Ren-chang  ZHENG Ying-hua
Abstract:Avermectin production of Streptomyces avermitilis could be stimulated by increasing the dosage of orfX gene. Integrative expression vector pWJ827 containing ermEp* and construction gene of orfX, was constructed based on pIJ8600. The recombinant strain BIB0827 was obtained by introducing pWJ827 into the industrial strain BIB9903 through intergeneric conjugal transfer. HPLC assays showed the avermectin production of BIB0827 reached 3350μg/mL, 1.5-fold increasing when compared with BIB9903. The characteristic of over producing strain BIB0827 was stable after ten rounds of non-selective growth, which could be applied in the industrial production of avermectin.
Keywords:orfX  conjugal transfer  integrative expression  Streptomyces avermitilis  avermectin
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