不吸水链霉菌武夷变种CK-15遗传转化体系的探索及初建  

Construction of the Conjual Transfer System of Streptomyces ahygroscopicus var.wuyiensis CK-15

作　　者：杨振娟[1] 孙蕾[1] 武哲[1] 张俊[2] 檀贝贝[1] 张克诚[1] 

机构地区：[1]中国农业科学院植物保护研究所、农业部作物有害生物综合治理重点实验室,北京100193 [2]河南农业大学植物保护学院,郑州450002

出　　处：《生物技术通报》2012年第10期193-198,共6页Biotechnology Bulletin

基　　金：公益性行业科研专项资金项目(201103002);中国农业科学院植物保护研究所中央级公益性科研院所基本科研业务费专项资金项目(1610142011004)

摘　　要：旨在对不吸水链霉菌武夷变种CK-15的遗传转化体系进行初步的探索及建立。首先对内源质粒的存在及对抗生素敏感性进行了测定,结果显示,在不吸水链霉菌武夷变种CK-15中不存在外源质粒。最终确定最适培养基为MS培养基,最适热激温度与时间为50℃热激10 min,37℃温育最适时间为2.5 h,最佳抗生素水溶液覆盖时间为16-18 h,并对转化子进行了验证。For the further molecular biological research,analysis and modification in gene level of wuyiensis,we construct a system of genetic transformation that is suitable for Strepomyces ahygroscopicus var.wuyiensis CK-15 in this study.We first test if the indigenous plasmid exits and the resistance detection.The result showed that there is no indigenous plasmid in Strepomyces ahygroscopicus var.wuyiensis CK-15.And the result also showed that MS medium is fit for conjugal transfer between CK-15 spores and E.coli ET12567（PUZ8002）,and the pre-conditions that the spores were heated at 50℃ for 10 min,incubated at 37℃ for 2.5 hours are needed.16-18 h after cultivated covered the medium with 1 mL water that contains antibiotic and Nalidixic acid.Then,we verified the transformants.

关 键 词：武夷菌素 接合转移 遗传转化 内源质粒 抗性检测 pKC1139 pSET152 

分 类 号：S476[农业科学—农业昆虫与害虫防治]