%0 Journal Article
%T Prokaryotic expression of the major antigenic domain of equine arteritis virus GL protein and the establishment of putative indirect ELISA assay<br>马动脉炎病毒GL蛋白主要抗原域的表达及间接ELISA的初步建立
%A LIANG Cheng-zhu
%A CAO Rui-bing
%A WEI Jian-chao
%A ZHU Lai-hua
%A CHEN Pu-yan
%A <br>梁成珠
%A 曹瑞兵
%A 魏建超
%A 朱来华
%A 陈溥言
%J 微生物学报
%D 2006
%I 
%X According to the antigenic analysis of equine arteritis virus (EAV) GL protein, one pair of primers were designed, with which the gene fragment coding the high antigenic domain of EAV GL protein was amplified from the EAV genome. The cloned gene was digested with BamH I and Xho I and then inserted into pET-32a and resulted pET-GL1. The pET-GL1 was transformed into the host cell BL21(DE3) and the expression was optimized including cultivation temperature and concentration of IPTG. The aim protein was highly expressed and the obtained recombinant protein manifested well reactiongenicity as was confirmed by Western blot. The recombinant GL1 protein was purified by the means of His * Bind resin protein purification procedure. Then an indirect ELISA was established to detect antibody against EAV with the purified GL1 protein as the coating antigen. The result showed that the optimal concentration of coated antigen was 9.65 microg/mL and the optimal dilution of serum was 1:80. The positive criterion of this ELISA assay is OD (the tested serum) > 0.4 and OD (the tested serum) /OD (the negative serum) > 2.0. The iGL-ELISA was evaluated versus micro-virus neutralization test. The ELISA was performed on 900 sera from which were preserved by this lab during horse entry/exit inspection, the agreement (94.1%) of these test were considered suitable for individual serological detection. In another test which 180 sera samples were detected by iGL-ELISA and INGEZIM ELISA kit respectively. The agreement ratio between the two methods is 95.6%.
%K Equine arteritis virus
%K GL protein
%K Major antigenic domain
%K Prokaryotic expression
%K Indirect ELISA<br>马动脉炎病毒
%K GL蛋白
%K 主要抗原域
%K 原核表达
%K 间接ELISA
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A3C6BA55AB623B90FA9104CFFC826F3C&aid=423EDC48C28AAA1C&yid=37904DC365DD7266&vid=D997634CFE9B6321&iid=38B194292C032A66&sid=08076B8B3CC96095&eid=78976D931AD1540F&journal_id=0001-6209&journal_name=微生物学报&referenced_num=4&reference_num=12