| PCR方法筛选淡紫灰链霉菌海南变种基因组粘粒文库 |
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| 引用本文： | 田云龙,朱昌雄,蒋细良,郭萍,刘雪.PCR方法筛选淡紫灰链霉菌海南变种基因组粘粒文库[J].生物技术通报,2009(11). |
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| 作者姓名： | 田云龙 朱昌雄 蒋细良 郭萍 刘雪 |
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| 作者单位： | 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 |
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| 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4.21×106CFU/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30～40 kb,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.
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| 关 键 词： | 中生菌素 生物合成基因 基因组 粘粒文库 筛选 |
The PCR-based Screening of the Cosmid Library from Streptomyces lavendulae var.hainanensis |
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| Tian Yunlong,Zhu Changxiong,Jiang Xiliang,Guo Ping,Liu Xue.The PCR-based Screening of the Cosmid Library from Streptomyces lavendulae var.hainanensis[J].Biotechnology Bulletin,2009(11). |
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| Authors: | Tian Yunlong Zhu Changxiong Jiang Xiliang Guo Ping Liu Xue |
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| Abstract: | The cosmid genomic library of Streptomyces lavendulae var. hainanensis B-7 producing zhongshengmycin was constructed by using pOJ446 as cosmid vector. The liter of the constructed cosmid library was 4. 21× 10~6 CFU/ml. 12 colonies were picked,and their comid vectors were prepared and digested with BamHI. As a result,the electrophoresis 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. |
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| Keywords: | Zhongshengmycin Biosynthetic genes Cenomic Cosmid library Screening |
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