大豆花叶病毒诱导的应用于膜蛋白酵母双杂交的大豆cDNA文库构建  

Construction of Soybean mosaic virus-induced soybean cDNA library for membrane-based yeast two-hybrid system

作　　者：崔晓艳[1] 陈新[1] 栾鹤翔[2] 智海剑[2] 

机构地区：[1]江苏省农业科学院蔬菜研究所,南京210014 [2]南京农业大学农学院,南京210095

出　　处：《植物病理学报》2013年第5期556-560,共5页Acta Phytopathologica Sinica

基　　金：国家自然科学基金(31101411);江苏省农业科技自主创新基金(CX(12)5045);转基因抗大豆花叶病毒病高蛋白大豆新品种的培育(2010ZX08004-004-004)

摘　　要：前言大豆花叶病毒（Soybean mosaic virus，SMV）是大豆上最普遍的病害之一，严重影响大豆的产量和品质，通过筛选与病毒互作的寄主蛋白，根据互作蛋白的功能和两者的作用机制可揭示病毒侵染的分子机制。对于病毒编码的膜蛋白或者膜相关特性的蛋白，由于具有膜结合特性，在生物体内的互作多发生在膜上。Membrane-baesd yeast two-hybrid system is an effective method for research on interaction between Soybean mosaic virus-encoded membrane-associated proteins and host factors, while the Gateway technology without the use of restriction enzyme cloning techniques is easier for construction of virus-induced host eDNA li- brary. In this study, both membrane-based yeast two-hybrid system and GatewayTM systems were used. With TRIZOL regent, total RNA was extracted from soybean leaves infected with soybean mosaic virus. SMV-in- duced soybean primary eDNA library constructed by Gateway technology was recombined into a reconstructed prey vector for membrane-based yeast two-hybrid system. The capacity and quality tests showed that the library titer was 1.7 × 10^6cfu/mL and the length of inserted eDNA fragments ranged from 0.5 to 2 kb. It is available for research on interaction between the virus-encoded membrane protein and host.

关 键 词：豆类作物 大豆花叶病毒诱导 膜蛋白酵母双杂交 大豆cDNA文库 

分 类 号：S52[农业科学—作物学]