| 玉米纹枯病菌全长cDNA文库的构建与鉴定 |
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| 引用本文： | 孔婷婷,高增贵,张硕,王敏,周艳波,王小皙.玉米纹枯病菌全长cDNA文库的构建与鉴定[J].植物病理学报,2014,44(6):603-608. |
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| 作者姓名： | 孔婷婷 高增贵 张硕 王敏 周艳波 王小皙 |
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| 作者单位： | 沈阳农业大学 植物免疫研究所, 沈阳 110866 |
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| 基金项目： | 国家科技支撑计划“粮食丰产科技工程”项目(2011BAD16B12;2012BAD04B03;2013BAD07B03);辽宁省玉米育种及配套技术研究创新团队项目(2014201003) |
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| 摘 要： | 以玉米纹枯病菌菌丝为材料,利用In-Fusion SMARTerTM cDNA Lib Construction Kit构建玉米纹枯病菌全长cDNA文库。文库质量鉴定结果表明,文库滴度为1.2×106pfu·mL-1,重组率为99.2%,插入片段平均长度大于1.0 kb。随机挑取400个白色克隆进行测序,共获得329个高质量EST序列,经聚类拼接后得到250条unique EST,包括36条contigs和214条singletons。在GenBank 进行Blastn 与Blastx 同源比对,有227条EST 与已知核酸或蛋白有不同程度的同源性,占全部EST 的90.8%,其余23条无任何同源性,占9.2%。
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| 关 键 词： | 玉米纹枯病菌 cDNA文库 表达序列标签(EST) |
| 收稿时间： | 2013-06-03 |
Construction and characterization of a full-length cDNA library of Rhizoctonia solani |
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| KONG Ting-ting,GAO Zeng-gui,ZHANG Shuo,WANG Min,ZHOU Yan-bo,WANG Xiao-xi.Construction and characterization of a full-length cDNA library of Rhizoctonia solani[J].Acta Phytopathologica Sinica,2014,44(6):603-608. |
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| Authors: | KONG Ting-ting GAO Zeng-gui ZHANG Shuo WANG Min ZHOU Yan-bo WANG Xiao-xi |
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| Institution: | Institute of Plant Immunology, Shenyang Agricultural University, Shenyang 110161, China |
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| Abstract: | A full-length cDNA library was constructed from hypha of Rhizoctonia solani by using In-Fusion SMARTerTM cDNA Library Construction Kit. The constructed library had a high titer of 1.2×106 pfu·mL-1 and a 99.2% recombination rate, of which the average length of the inserted cDNA fragment was above 1.0 kb. The 329 high quality ESTs were obtained from 400 white clones picked randomly. After treatments of clustering and splicing, 250 unique ESTs were obtained, including 36 contings and 214 singlets. These unique ESTs were analyzed for sequence homology with Blastx and Blastn software against sequences in the GenBank. Out of the 250 unique ESTs , 227 ESTs that accounted for 90.8% of all the unigenes showed homology with the function known genes or proteins,whereas the rest that accounted for 9.2% has no homology. |
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| Keywords: | Rhizoctonia solani cDNA library expressed sequence tag ( EST) |
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