中国农业科学院农业信息研究所 中国农学会
Agricultural Information Institute of CAAS China Association of Agricultural Science Societies
En
Ch
中国农业科学院农业信息研究所 中国农学会 主办

AI检索

订阅

返回顶部
当前位置: 首页 > 文章 > 一种快速高效构建植物表达载体的方法 玉米科学 2012,20 (1) 61-66
Position: Home > Articles > A Rapid and High Efficient Method to Construct Plant Expression Vectors Journal of Maize Sciences 2012,20 (1) 61-66

一种快速高效构建植物表达载体的方法

作  者：
韩凯;翁建峰;郝转芳;李新海;李明顺;张德贵;白丽;张世煌;薛吉全;谢传晓
单  位：
中国农业科学院作物科学研究所/农作物基因资源与基因改良国家重大科学工程;西北农林科技大学农学院
关键词：
表达载体;同源序列;DNA重组
摘  要：
植物表达载体是将目的基因转化宿主细胞的媒介,载体构建是通过遗传转化方法开展基因功能鉴定与应用等研究的重要环节。以构建玉米谷氨酰胺合成酶基因(GS1)表达载体pCUbi1390-Ubi-GS1-35S-EPSPS为例,研究简单、通用、高效的构建载体的方法。该方法目的片段和载体不需要酶切产生互补的黏性末端,只需通过在目的基因PCR上下游引物的5’端设计与线性载体两端有15个碱基的同源序列,根据序列的同源性,将目的基因插入载体中,通过PCR程序实现DNA重组。结果表明,正确构建了设计的转化载体,该方法大大简化了载体构建的过程,也适用于任何一个基于单酶切位点把外源DNA重组插入目标序列。
译  名：
A Rapid and High Efficient Method to Construct Plant Expression Vectors
作  者：
HAN Kai1,2,WENG Jian-feng2,XUE Ji-quan1,XIE Chuan-xiao2,et al.(1.College of Agriculture,Northwest A&F University,Yangling 712100; 2.Institute of Crop Science,Chinese Academy of Agricultural Sciences / Crop Gene Resources and Genetic Improvement,National Key Science and Engineering,Beijing 100081,China)
关键词：
Expression vectors;Homology;DNA recombination
摘  要：
The plant expression vector is the vehicle that takes target gene into and then expresses the target gene in the host cells.Therefore,expression vector construction is one of the key steps of theoretical and applied studies through genetic modification strategy.The demonstration of constructing GS1 gene,encoding a cytosolic glutamine synthetase,into pCUbi1390-Ubi-GS1-35S-EPSPS,a rapid,simple and high efficient constructing method had been introduced in this paper.After a vector linear step with single restriction digestion reaction,the homolog recombination could be executed through PCR reaction,in which the PCR primers for the target insert sequence were designed to share 15 bases of homology with the sequences at expecting site the end of the linearized vector.It was unnecessary to digest the target gene and vector to produce complementary cohesive ends in this method.The results showed the expecting pCUbi1390-Ubi-GS1-35S-EPSPS was successfully constructed.The procedures were largely simplified,and the method could be widely used in any recombination of insertion of any DNA sequence not very long at the single restriction site of any DNA sequence.

相似文章

导出引用
计量
文章访问数: 9
HTML全文浏览量: 0
PDF下载量: 0

所属期刊

推荐期刊

友情链接Friendly link
农业信息研究所Agricultural information institute 农业智能知识服务平台Agricultural intelligent knowledge service platform 农业学术服务平台Agricultural Knowledge Service System 中国知网CNKI 维普网VIP 万方WANFANG DATA
联系我们Contact us

电话Tell：010-82109652-808

邮箱E-mail：agrijournal@caas.cn

服务Service
关于我们About us 意见建议Opinions and suggest
版权说明Copyright
单位：中国农业科学院农业信息研究所 中国农学会 © 2017中国农业科学院农业信息研究所 ICP备案号：京ICP备09089781号-46