ELISA验证兔铁蛋白重链与RHDV VP60相互作用  

ELISA verifies the interaction between Ferritin heavy chain and VP60 of rabbit hemorrhagic disease virus

作　　者：孙宏勇[1] 刘家森[1] 王泽祥[1] 陈淑慧[1] 单丽玲[1] 郭冬春[1] 姜骞[1] 林欢[1] 曲连东[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/实验动物研究室,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2013年第9期770-772,共3页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金(30800830);黑龙江省科技计划项目(PC10S02)

摘　　要：为验证本实验室前期实验中初步鉴定的兔铁蛋白重链(FTH)与兔出血症病毒(RHDV)衣壳蛋白(VP60)具有相互作用关系,本研究采用RT-PCR技术从兔肝脏扩增编码FTH基因,并将其克隆至pET-32a(+)中转化大肠杆菌BL21(DE3)pLysS中进行表达。重组FTH蛋白(rFTH)经IPTG诱导获得表达,采用Ni-NTA Agarose纯化rFTH。利用纯化的VP60作为ELISA包被抗原检测FTH与VP60的相互作用。结果表明,ELISA检测的OD490nm值与FTH蛋白量呈正相关。本研究进一步验证了FTH与VP60具有相互作用关系。To verify the interaction between Ferritin heavy chain （FTH） and rabbit hemorrhagic disease virus （RHDV） VP60 which was preliminarily identified in our previous experiment, the FTH gene were amplified by RT-PCR from total RNA of rabbit hepatic cells and cloned into pET32a（＋） vector to express in E.coli. The recombinant protein （rFTH） was expressed with IPTG induction and purified by Ni-NTA agarose. Furthermore, the interaction between FTH and VP60 was detected by ELISA coating with VP60 as the capture antigen for detection of rFTH. The result showed that OD490nm value increased following the increasing concentration of rFTH, which showed in a concentration-dependent manner. These data further confirmed that the FTH was interacted with RHDV VP60.

关 键 词：铁蛋白重链 兔出血症病毒 VP60 ELISA 

分 类 号：S852.65[农业科学—基础兽医学]