CHD9:一个新的与流感病毒PA蛋白相互作用的宿主因子

PA蛋白是禽流感病毒(AⅣ)聚合酶的组成部分,对AIV的复制具有重要作用.为鉴定家鸭体内与AIV PA蛋白相互作用的宿主蛋白,本研究利用酵母双杂交系统,以表达A/goose/Hubei/65/05(H5N1)AIV PA蛋白的重组质粒为诱饵,筛选家鸭脑组织cDNA文库,鉴定得到一个cDNA片段,经测序分析表明,该cDNA片段编码蛋白为色素域解旋酶DNA结合蛋白9(CHD9).将重组诱饵质粒pDEST32-PA与捕获的鸭cDNA重组质粒共转化MaV203受体菌,进行X-gal显色反应检测,分析结果显示,菌落可以迅速变蓝,并且菌落在SC-Leu-TrpHis+3AT 3种营养缺陷培养基平板上生长良好.证明CHD9与PA蛋白在酵母双杂交系统中具有较强的相互作用.CHD9的初步鉴定为研究AIV在机体中的复制过程奠定了基础.

CHD9,a novel host factor interacting with PA protein of influenza virus

PA protein is a component of influenza virus polymerase,which plays an important role in the influenza virus replication.In this study,we screened domestic ducks cerebral cDNA library with the recombinant plasmid(pDEST32-PA) expressing PA protein of A/goose/Hubei/65/05 avian influenza virus(AIV) as bait by yeast two-hybrid system.As a result,we identified a cDNA fragment,which encoding chromodomain helicase DNA-binding protein9(CHD9).Then the recombinant baitplasmid and the prey recombinant plasmid containing the cDNA fragment of CHD9 were co-transformed into the yeast strain MaV203.The X-gal analysis shown that the yeast colonies were quickly turned into blue and the recombinant yeast grown well in SC-Leu-Trp-His 3AT plate,which demonstrated the CHD9 from duck was able to strongly interact with AIV PA protein.The primary identification of the interaction between CHD9 and PA protein needed further validation in other systems to provide research-based to clarify the role of CHD9 in influenza virus replication process.