与水稻黑条矮缩病毒p5b互作的水稻基因片段筛选

 由水稻黑条矮缩病毒基因组S5片段第2个开放阅读框（ORF）编码的p5b是一个功能未知的病毒非结构蛋白。构建了酵母双杂交体系的含水稻cDNA表达文库的pGADT7载体，同时将p5b 基因构建到诱饵质粒pGBKT7上（pGBK p5b），Western Blot和自激活实验结果显示p5b可在酵母中正确表达且无自激活活性。进一步利用p5b蛋白作为诱饵用酵母双杂交筛选水稻cDNA表达文库，鉴定出一些参与光合作用、呼吸作用等重要代谢过程的关键酶基因片段，这些酶可能与p5b蛋白之间存在相互作用，推测这些互作在病株症状发展过程中起作用。

Screening of Rice Genes Interacted with p5b of Rice black-streaked dwarf virus

The second open reading frame on the genome segment 5 (S5) of Rice black streaked dwarf virus (RBSDV) encodes a viral nonstructural protein (p5b) with unknown function. To reveal the function of p5b, its gene was ligated into bait plasmid pGBKT7 and an expression library containing rice cDNAs was constructed using plasmid pGADT7 of yeast two hybrid. The bait protein p5b was detected in yeast by western blot and the result of an auto activation test showed that p5b could not autonomously activate the expression of reporter genes in yeast. Then the bait protein p5b was used for screening the expression cDNA library of rice plant. Gene fragments of some pivotal enzymes, involved in photosynthesis, respiration, and other important metabolism processes, were identified to interact with p5b in yeast, suggesting that these interactions may play roles in symptom development of infected plants.