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大豆Kunitz型胰蛋白酶抑制剂基因RNAi载体的构建 预览 被引量:3

Construction of RNAi Expression Vector of Kunitz Trypsin Inhibitor Gene from Soybean
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摘要 采用PCR技术扩增得到胰蛋白酶抑制剂KTi基因正义和反义片段、大豆种子特异性启动子7αP和作为内含子的GUS基因片段,将其分别连入克隆载体pMD18-TVector中。然后以植物表达载体pCAMBIA1301为基础,通过AHLG作为中间载体,根据RNAi原理将组成RNAi结构的4个目的片段分别连入其中,成功构建了种子特异性RNAi表达载体p130t—KTiRi。研究结果为RNAi技术在大豆品质改良中的应用提供了基础。 In this research, the RNAi expression vector of Kunitz Trypsin inhibitor gene from soybean was constructed with inverted repeats of KTi gene, for suppressing the expression of KTi gene in soybean. The four gene fragments of RNAi struc- ture including the sense and anti-sense fragments of KTi gene, soybean seed-specific promoter 7αp and the fragment of GUS gene which used as intron were cloned by PCR method, and then inserted into pMD18-T Vector respectively to get four recombinant cloned vectors. Then the RNAi expression vector pl301-KTiRi was constructed based on the vector of pCAM- BIA130lwith AHLG as the bridging vector. This study provides the basis for improving the quality of soybean with RNAi method.
作者 周海涛 付永平 王丕武 ZHOU Hai-tao, FU Yong-ping, WANG Pi-wu (College of Agriculture, Jilin Agricultural University, Changchun 130118, Jilin, China)
出处 《大豆科学》 CSCD 北大核心 2010年第5期 742-746,共5页 Soybean Science
基金 基金项目:国家自然科学基金资助项目(30971805).
关键词 大豆 Kunitz型胰蛋白酶抑制剂 RNA干扰 Soybean Kunitz trypsin inhibitor RNA interference
作者简介 第一作者简介:周海涛(1983-),男,在读硕士,研究方向为作物遗传育种。E—mail:show19830623@yahoo.com.cn。 通讯作者:王丕武,教授,博士生导师。E-mail:peiwuw@yahoo.com.cn。
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