PCR方法筛选淡紫灰链霉菌海南变种基因组粘粒文库  

The PCR-based Screening of the Cosmid Library from Streptomyces lavendulae var. hainanensis

作　　者：田云龙[1] 朱昌雄[1] 蒋细良[2] 郭萍[1] 刘雪[1] 

机构地区：[1]中国农业科学院农业环境与可持续发展研究所,北京100081 [2]中国农业科学院植物保护研究所,北京100081

出　　处：《生物技术通报》2009年第11期117-120,共4页Biotechnology Bulletin

基　　金：国家"863"项目(2006AA10A209)

摘　　要：以中生菌素产生菌淡紫灰链霉菌海南变种（Streptomyces lavendulae var.hainanensis）B-7菌株为材料,利用pOJ446作为载体构建了B-7菌株的基因组粘粒文库,文库效价达到4.21×10^6CFU/ml。随机挑取12个克隆提取质粒,经BamHI酶切电泳分析,插入片段大小约为30-40kb,符合建库要求的理论值。利用建立的基于96深孔板PCR技术的文库筛选改良方法,成功快速地筛选到含有目标基因片段的阳性菌株。The cosmid genomic library of Streptomyces lavendulae var. hainanensis B-7 producing zhongshengmycin was constructed by using pOJ446 as cosmid vector. The titer of the constructed cosmid library was 4.21 × 10^6CFU/ml. 12 colonies were picked,and their comid vectors were prepared and digested with BamHI. As a result,the eleetrophoresis showed that the inserting DNA sizes ligated into the vectors were 30 kb to 40 kb which could fulfill the research mission. A PCR-based method for screening the cosmid library was developed and four clones which carried target DNA sequence were successfully screened.

关 键 词：中生菌素 生物合成基因 基因组 粘粒文库 筛选 

分 类 号：S476.1[农业科学—农业昆虫与害虫防治]