| 玉米MAPK5基因片段的克隆及RNA干扰载体的构建 |
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| 引用本文： | 崔喜艳,刘晶,丁志鑫,郝东云.玉米MAPK5基因片段的克隆及RNA干扰载体的构建[J].吉林农业大学学报,2012(4):368-372. |
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| 作者姓名： | 崔喜艳 刘晶 丁志鑫 郝东云 |
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| 作者单位： | 中国农业科技东北创新中心;吉林农业大学生命科学学院 |
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| 基金项目： | 吉林省博士后科研项目启动经费资助[吉人社办字(2009)第100号];吉林省科技发展计划项目(20086029) |
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| 摘 要： | 采用RT-PCR方法从玉米品种B73幼苗中克隆出促分裂原活化蛋白激酶基因5(MAPK5)的cDNA部分片段(691 bp),与GenBank已发表的玉米MAPK5(AB016802)同源性达97%。重新设计带酶切位点的引物,克隆出长度为281 bp的正、反向片段。使用DNA重组技术,用pCAMBIA3300质粒构建了玉米MAPK5的RNAi载体。
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| 关 键 词： | 促分裂原活化蛋白激酶 基因克隆 干扰载体 |
Cloning of Maize MAPK5 Gene Fragment and Its Construction of RNA Interfering Vector |
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| CUI Xi-yan,LIU Jing,DING Zhi-xin,HAO Dong-yun.Cloning of Maize MAPK5 Gene Fragment and Its Construction of RNA Interfering Vector[J].Journal of Jilin Agricultural University,2012(4):368-372. |
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| Authors: | CUI Xi-yan LIU Jing DING Zhi-xin HAO Dong-yun |
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| Institution: | 1.Northeast Agricultural Research Center of China,Changchun 130124,China;2.College of Life Sciences,Jilin Agricultural University,Changchun 130118,China |
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| Abstract: | The partial cDNA(691 bp)of mitogen-activated protein kinase five(MAPK5) gene was cloned from seeding of maize cultivar B73 by RT-PCR.Sequence homology analysis showed that it shared the high similarity(97%) with maize MAPK5 that has been submitted to GenBank(AB016802).To construct the RNAi vector of Maize MAPK5,primers with restriction endonuclease recognition sites were designed to amplify the 281 bp forward and reverse fragments of MAPK5,and the resulting PCR products were then inserted into the plasmid pCAMBIA3300 by DNA recombinant technology.This RNAi vector lays a foundation for further study of MAPK5 involved in MAPK cascades pathway of maize. |
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| Keywords: | mitogen-activated protein kinase gene cloning interference vector |
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