猪繁殖与呼吸综合征病毒N蛋白与BCCIP蛋白相互作用的鉴定  

Identification of the interaction between porcine reproductive and respiratory syndrome virus N protein and porcine BCCIP

作　　者：尹曼曼[1] 郭东伟[1] 刘琴芳[1] 张枫[1] 张利杰[1] 李江南[1] 翁长江[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/猪传染病研究室,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2013年第5期341-345,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金面上项目(31172333)

摘　　要：猪繁殖与呼吸综合征病毒(PRRSV)核衣壳(N)蛋白在病毒粒子组装和释放过程中发挥重要的作用。为筛选N蛋白与宿主细胞相互作用的蛋白,本研究分离健康猪肺泡巨噬细胞(PAM),构建了PAM的酵母cDNA文库,利用酵母双杂交方法筛选其中与PRRSV N蛋白相互作用的蛋白,结果获得3个阳性克隆。经测序并利用Blast方法比对GenBank数据库,筛选得到克隆插入的cDNA均与BCCIP(BRCA2 and CDKN1A interactingprotein)基因的1 bp～396 bp核酸序列一致。经酵母回返验证试验、GST-pulldown和免疫共沉淀试验验证了N蛋白与BCCIP之间的特异性相互作用,共定位试验显示N蛋白与BCCIP共定位于细胞核和细胞浆中。本研究鉴定了新的与PRRSV N蛋白相互作用的宿主蛋白BCCIP,BCCIP是否通过与N蛋白的相互作用参与PRRSV复制还有待深入研究。Porcine reproductive and respiratory syndrome virus （PRRSV） nuclear capsid （N） protein play an important role in the process of virus particle assembly and release. To screen the host proteins interacted with N protein, porcine alveolar macrophage （PAM） cDNA library was constructed from health swine PAM and screened the library using PRRSV N protein as ＂bait＂ in yeast two-hybrid system. Three positive clones were obtained. The inserted cDNAs were sequenced and analyzed with Blast assay, which encode the amino acid 1 aa-132 aa of porcine BCCIP. The interaction was further confirmed by the retest in yeast, GST pull-down, co-immunoprecipitation and colocalization in HEK293 cell. These results demonstrate that PRRSV N interacted with BCCIP. However, whether BCCIP involves in PRRSV infection and replication stilly need further study.

关 键 词：猪繁殖与呼吸综合征病毒 N蛋白 BCCIP 相互作用 

分 类 号：S852.65[农业科学—基础兽医学]