| 玉米弯孢叶斑病菌酵母双杂交cDNA 文库的构建及评价 |
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| 引用本文： | 荆晶,刘铜,陈捷.玉米弯孢叶斑病菌酵母双杂交cDNA 文库的构建及评价[J].植物病理学报,2012,42(1):93-96. |
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| 作者姓名： | 荆晶 刘铜 陈捷 |
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| 作者单位： | 1上海交通大学农业与生物学院农业部都市农业(南方)重点开放实验室, 上海200240;2 黑龙江八一农垦大学农学院, 大庆163319 |
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| 基金项目： | 国家自然科学基金(30871610;30971949);国家博士点专项基金(20090073110048);现代农业产业技术体系专项资金资助(CARS-02) |
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| 摘 要： | BD SMART technique and LD-PCR were applied to synthesize double strand cDNA(ds cDNA).The ds cDNA and pGADT7-Rec were transfered into competent yeast cell to construct yeast two-hybrid cDNA library of Curvularia lunata by homologous recombination method. The results showed that library capacitance was 2. 46 × 105 and transformation rate was 2. 13 × 105 / μg pGADT7-Rec. The plaque titer of the library was 2. 5 × 108 pfu / mL and the recombination frequency was 88. 24% . The length of inserted cDNA fragment was ranged from 0. 4 kb to 3 kb in average. This is the first time to use BD SMART technique to construct yeast two-hybrid cDNA library of C. lunata by homologous recombination.
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| 关 键 词： | Curvularia lunata yeast two-hybrid cDNA library |
Construction and characterization of yeast two hybrid cDNA library of Curvularia lunata |
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| JING Jing
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LIU Tong
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CHEN Jie.Construction and characterization of yeast two hybrid cDNA library of Curvularia lunata[J].Acta Phytopathologica Sinica,2012,42(1):93-96. |
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| Authors: | JING Jing LIU Tong CHEN Jie |
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| Institution: | 1 School of Agriculture and Biology, Shanghai Jiaotong University,Key Laboratory of Urban Agriculture(South), Ministry of Agriculture, Shanghai 200240, China; 2 School of Agriculture, Heilongjiang Bayi Agricultural University, Daqing 163319, China |
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| Abstract: | BD SMART technique and LD-PCR were applied to synthesize double strand cDNA(ds cDNA).The ds cDNA and pGADT7-Rec were transfered into competent yeast cell to construct yeast two-hybrid cDNA library of Curvularia lunata by homologous recombination method.The results showed that library capacitance was 2.46×105 and transformation rate was 2.13×105/μg pGADT7-Rec.The plaque titer of the library was 2.5×108 pfu/mL and the recombination frequency was 88.24%.The length of inserted cDNA fragment was ranged from 0.4 kb to 3 kb in average.This is the first time to use BD SMART technique to construct yeast two-hybrid cDNA library of C.lunata by homologous recombination. |
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| Keywords: | Curvularia lunata yeast two-hybrid cDNA library |
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