小麦AGPase基因LSU Ⅱ的克隆及反义表达与RNAi干扰载体的构建

doi:10.7668/hbnxb.2009.04.001

华北农学报 ›› 2009, Vol. 24 ›› Issue (4): 1-6. doi: 10.7668/hbnxb.2009.04.001

所属专题： 小麦；生物技术；

• 论文 • 下一篇

小麦AGPase基因LSU Ⅱ的克隆及反义表达与RNAi干扰载体的构建

刘超, 康国章, 郭天财, 岳彩风, 沈丙权

河南农业大学, 国家小麦工程技术研究中心, 河南郑州 450002

收稿日期:2008-08-19 出版日期:2009-08-28
通讯作者: 郭天财(1953-),男,河南济源人,博士生导师,主要从事作物生理方面的研究。
作者简介:刘超(1982-),男,河南新郑人,在读硕士,主要从事作物分子生理方面的研究.
基金资助:
国家"十五"科技攻关重大项目;河南省教育厅自然科学基金资助项目(2006210007)

Molecular Cloning of Plastidial Large Subunit Ⅱ Gene and Its Constructions of Antisense and RNAi Expression Vectors

LIU Chao, KANG Guo-zhang, GUO Tian-cai, YUE Cai-feng, SHEN Bing-quan

Wheat Engineering Research Center, Henan Agricultural University, Zhengzhou 450002, China

Received:2008-08-19 Published:2009-08-28

摘要/Abstract

摘要： 采用RT-PCR方法从小麦品种豫教2号的发育籽粒中克隆出小麦淀粉合成关键酶-AGPase质体型大亚基(AGPase plastidial large subunit,LSU Ⅱ)cDNA一段特异保守序列,长度为535 bp(GenBank No.EF378944),与DQ409820(小麦)、U87(大麦)、AK0929(水稻)和DQ40819(玉米)分别有98%,97%,88%,88%的同源性,表明克隆出了小麦LSU Ⅱ基因的部分cDNA序列.以pWM101质粒表达载体为基础,将LSU Ⅱ反向置于pWM101质粒的CaMV35S启动子之后,构建了LSU Ⅱ的反义表达载体pWM101LSU ⅡA;另外,用pFGC5941干扰载体构建了LSU Ⅱ基因的RNAi载体pFGC5941 LSU ⅡSA,这些载体的构建为研究LSU Ⅱ基因的功能打下了基础.

关键词: 小麦, AGPase, LSU Ⅱ, 表达载体

Abstract: Partial cDNA fragment (535 bp) of plastidial large subunit of AGPase (LSU II) from grains of common wheat variety Yujiao 2 was cloned (GenBank No. EF378944). The sequence analysis demonstrated that the cloned LSU II shared 88 % - 98 % homogeneity with the sequences of DQ409820,U66876,AK069296,DQ406819 genes. The antisense expression vector (pWM101LSU IIA) was constructed with pWM101 plasmids,and RNAi vector (pFGC5941LSU IISA) was also constructed with pFGC5941. The constructed vectors provided a good background for studying the function of LSU II.

Key words: Wheat ( Triticum aestivum), AGPase, LSU II, Plant expressing vectors

中图分类号:

Q785

刘超, 康国章, 郭天财, 岳彩风, 沈丙权. 小麦AGPase基因LSU Ⅱ的克隆及反义表达与RNAi干扰载体的构建[J]. 华北农学报, 2009, 24(4): 1-6. doi: 10.7668/hbnxb.2009.04.001.

LIU Chao, KANG Guo-zhang, GUO Tian-cai, YUE Cai-feng, SHEN Bing-quan. Molecular Cloning of Plastidial Large Subunit Ⅱ Gene and Its Constructions of Antisense and RNAi Expression Vectors[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2009, 24(4): 1-6. doi: 10.7668/hbnxb.2009.04.001.

http://www.hbnxb.net/CN/Y2009/V24/I4/1

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小麦AGPase基因LSU Ⅱ的克隆及反义表达与RNAi干扰载体的构建

Molecular Cloning of Plastidial Large Subunit Ⅱ Gene and Its Constructions of Antisense and RNAi Expression Vectors

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小麦AGPase基因LSU Ⅱ的克隆及反义表达与RNAi干扰载体的构建

Molecular Cloning of Plastidial Large Subunit Ⅱ Gene and Its Constructions of Antisense and RNAi Expression Vectors

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