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

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 cDNA library. In this study, both membrane-based yeast two-hybrid system and Gateway^TM systems were used. With TRIZOL regent, total RNA was extracted from soybean leaves infected with soybean mosaic virus. SMV-induced soybean primary cDNA 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 ? 10⁶cfu/mL and the length of inserted cDNA fragments ranged from 0.5 to 2 kb. It is available for research on interaction between the virus-encoded membrane protein and host.