中国水稻科学

中国水稻科学

• 研究报告 • 上一篇    下一篇

以cosmid质粒为载体的稻瘟病菌转化体系的建立及病菌基因文库的构建

鲁国东1; 黄大年2; 陶全洲3; 杨炜2; 谢联辉1   

  1. 1 福建农业大学植物保护系，福州350002；2 中国水稻研究所生物工程系，杭州310006；3 中国科学院上海生物化学研究所，上海200031
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:1995-07-10 发布日期:1995-07-10

Establishment of Transformation System and Genomic DNA Library of Magnaporthe grisea with Cosmid Vector

Lu Guodong; Huang Danian; Tao Quanzhou; Yang Wei; Xie Lianhui   

  • Received:1900-01-01 Revised:1900-01-01 Online:1995-07-10 Published:1995-07-10

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摘要： 用潮霉素B（ hygromycin B）抗性基因和λ噬菌体的cos位点组建了一种cosmid质粒pSVhygB，以此质粒建立了稻瘟病菌的转化体系，并且构建了病菌的基因组文库。结果表明，pSVhygB质粒可用于稳定地转化稻瘟病菌，转化频率约为15个转化子／μg DNA，电击法转化并不能有效地提高稻瘟病菌的转化频率。构建的稻瘟病基因文库中包含将近10000个重组子克隆，随机挑取12个重组子的鉴定表明，都包含有外源插入片段，说明所建文库已达要求。

关键词: cosmid质粒, 基因组文库, 潮霉素B抗性标记

Abstract: A new cosmid vector pSVhygB containing hygromycin B resistance gene and bacterlophage λcos site was constructed. Using the pSVhygB the genomic DNA library of Magnaporthe grisea was constructed. It showed that pSVhygB can stably transform M. grisea with frequency 15 transformants per microgram DNA and electroporation could not improved the transformation efficiency. There approximately had 10,000 recombinant clones in our genomic DNA library, more than that of the requirement for M. grisea.

Key words: cosmid vector, genomic DNA library, hygromycin B resistant marker, Magnaporthe grisea, transformation