| Gateway技术构建交链孢菌JH505 cDNA文库 |
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| 引用本文： | 龙松华 张宁 邱德文,黎定军 黄炜.Gateway技术构建交链孢菌JH505 cDNA文库[J].微生物学报,2005,45(6):963-965. |
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| 作者姓名： | 龙松华 张宁 邱德文 黎定军 黄炜 |
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| 作者单位： | 龙松华（中国农业科学院农业环境与可持续发展研究所,北京,10081;湖南农业大学生物安全科技学院,长沙,410128）;张宁（中国农业科学院农业环境与可持续发展研究所,北京,10081）;邱德文（中国农业科学院农业环境与可持续发展研究所,北京,10081）;黎定军（湖南农业大学生物安全科技学院,长沙,410128）;黄炜（中国农业科学院农业环境与可持续发展研究所,北京,10081） |
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| 基金项目： | 国家"973项目"(2003CB114204) |
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| 摘 要： | Gateway(R)技术构建Cdna文库,利用λ噬菌体的位点特异性重组,避免使用限制性内酶切割Cdna,能够解决常规方法构建Cdna文库的技术缺陷.首次应用Gateway(R)技术构建交链孢菌Cdna文库,经检测Cdna入门文库的滴度达到1×107cfu/Ml,文库总容量为9×107cfu,平均插入片段为1510bp.通过LR重组把入门文库转换为表达文库,表达文库的滴度为1.58×106cfu/Ml,文库总容量为6.32×106cfu,平均插入片段大小为1680bp.表达文库的构建为进一步克隆植物激活蛋白基因打下了基础.
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| 关 键 词： | Gateway技术 交链孢菌 激活蛋白 cDNA入门文库 cDNA表达文库 |
| 文章编号： | 0001-6209(2005)06-0963-03 |
| 修稿时间： | 2005年4月5日 |
Construction of a cDNA
library of Alternaria sp. strain JH505 using Gateway technology |
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| LONG Song-hua.Construction of a cDNA
library of Alternaria sp. strain JH505 using Gateway technology[J].Acta Microbiologica Sinica,2005,45(6):963-965. |
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| Authors: | LONG Song-hua |
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| Institution: | Institute of Environment and Sustainable Development in Agriculture, Chinese Academy of Agricultural Sciences, Beijing 100081, China. longsonghua79452@sohu.com |
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| Abstract: | A cDNA library was constructed based on the Gateway system that is a method to construct a high-quality cDNA library without the use of restriction enzyme for cloning. This is the first report that the Gateway system is used to construct a cDNA library for Alternaria sp.. The entry cDNA library was constructed in this report has a high titer of 1 x 10(7) cfu/mL and contain a total clones of 9 x 10(7) cfu, with an average inserts size of about 1510bp. In order to screen for a gene encoding the plant activator protein from library using an antiserum, the entry cDNA library was transferred into Gateway destination vector to create an expression library. The expression library show a high titer of 1.58 x 10(6) cfu/mL and contains a total clones of 6.32 x 10(6) cfu, with an average inserts size of 1680bp. |
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