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

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.