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中国农学通报

中国农学通报 ›› 2009, Vol. 25 ›› Issue (12): 25-29.

所属专题： 生物技术

• 生物技术科学 • 上一篇    下一篇

链格孢菌（Alternaria tenuissima）cDNA酵母表达文库的构建

冯飞,梁佳勇,纪春艳,曾洪梅,邱德文   

  • 收稿日期:2009-02-11 修回日期:2009-04-03 出版日期:2009-06-20 发布日期:2009-06-20

Construction of cDNA yeast expression library of Alternaria tenuissima

Feng Fei, Liang Jiayong, Ji Chunyan, Zeng Hongmei, Qiu Dewen   

  • Received:2009-02-11 Revised:2009-04-03 Online:2009-06-20 Published:2009-06-20

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摘要： 为了解链格孢菌（Alternaria tenuissima）遗传学背景，克隆及研究该菌功能基因，根据Gateway技术构建了既能在酵母细胞中表达外源基因又能在原核细胞大量复制的酵母表达载体pRS-DEST42，构建了细极链格孢菌（A. tenuissima）cDNA酵母表达文库。经检测，该文库的平均滴度为2.44×106（cfu/ml），文库总容量为2.44×107，阳性克性率为100%，平均插入片段约为1.38kb。细极链格孢菌（A. tenuissima）cDNA酵母表达文库是一个质量较高的表达文库，为克隆与分离全长目的基因及其功能奠定了坚实的基础。

关键词: 山区, 山区, 植被恢复, 进展

Abstract: For further understand genetics background and functional genes of A. tenuissima. A yeast destination vector pRS-DEST42, express foreign genes in a low-copy number in yeast and high-copy in E. coli, was constructed. The entry cDNA library of A. tenuissima was transferred into vector pRS-DEST42 through Gateway system. The cDNA yeast expression library has a higher titer of 2.44×106(cfu/ml) and a total clones of 2.44×107. The rate of positive recombinants clones was 100% and the size of average insert cDNA was 1.38 kb. This provides a basis for isolate and study on functional genes in A. tenuissima.