CAJ | 학술논문

为筛选人脑cD N A文库中与狂犬病病毒基质蛋白（M 蛋白）具有相互作用的蛋白基因，并对阳性克隆进行初步的分析和鉴定。首先构建了含有M 基因的诱饵质粒pGBK T7-CVS11-M ，并对其进行自激活和毒性检测，然后采用酵母双杂交技术初步筛选出与M 蛋白具有相互作用的12个阳性克隆，对阳性克隆进行PC R鉴定及测序分析，并通过酵母回复性试验最终获得3个无重复性克隆，编码的蛋白质分别为TAB2、P4HA2和RCN1。为研究狂犬病病毒M 蛋白在致病机制中的作用提供了参考。
위사선인뇌cD N A문고중여광견병병독기질단백（M 단백）구유상호작용적단백기인，병대양성극륭진행초보적분석화감정。수선구건료함유M 기인적유이질립pGBK T7-CVS11-M ，병대기진행자격활화독성검측，연후채용효모쌍잡교기술초보사선출여M 단백구유상호작용적12개양성극륭，대양성극륭진행PC R감정급측서분석，병통과효모회복성시험최종획득3개무중복성극륭，편마적단백질분별위TAB2、P4HA2화RCN1。위연구광견병병독M 단백재치병궤제중적작용제공료삼고。
In this study ,a human brain cDNA library was screened to identify rabies virus matrix protein (M protein) interaction partners ,and the positive clones were preliminarily identified .The bait plasmid pGBKT7-CVS11-M was constructed successfully and tested for auto-activation and toxicity and then was used to screen the proteins interacting with M protein in a yeast two-hybrid assay .Twelve positive clones were obtained .Subsequently ,they were verified by PCR and sequencing analysis ,and further conformed by retransformation assay .Three final positive proteins were verified as interacting proteins with M pro-tein through retransformation assay in yeast cells ,which were TAB2 ,P4HA2 and RCN1 .The research will contribute to explore the function of M protein at molecular level ,which would provide important clues for studying the pathogenic mechanism of rabies virus .