%0 Journal Article
%T Cloning,Prokaryotic Expression of CSFV NS3 Gene,and Preliminary Establishment of an Indirect ELISA for Serum Antibody Detection<br>猪瘟病毒NS3基因克隆、原核表达及间接ELISA方法初步建立
%A JIANG Da-Liang YU Xing-Long LI Run-Cheng GE Meng LUO Wei YAN Ai LI Jie LIU Chun-Hong TU Chang-Chun
%A <br>蒋大良
%A 余兴龙
%A 李润成
%A 葛猛
%A 罗维
%A 颜爱
%A 李杰
%A 刘春红
%A 涂长春
%J 微生物学通报
%D 2010
%I 
%X NS3 gene fragment (about 2000 bp) was amplified by PCR from plasmid of pPOHCLV containing Hog Cholera Lapinized Virus (HCLV) cDNA, and cloned into the prokaryotic expression vector pET-32a (+) to obtain the recombinant prokaryotic expression vector pETNS3. The pETNS3 was transformed into E.coli Rosetta (DE3) and expressed optimally. The recombinant protein NS3 about 95 kD was detected by SDS-PAGE and expressed mainly in the form of inclusion bodies. The result of Western blotting showed recombinant protein NS3 has immunogenicity. The nickel affinity chromatography was employed to purify the target protein, and purified recombinant protein NS3 (90%) was obtained. An indirect ELISA was initially established to detect antibody against CSFV NS3 protein, and 221 sera samples of pig from different herds and ages were tested. The result was compared with CSFV-Ab test kit of IDEXX, and the result showed: the positive coincidence rate was 83.33%, the negative coincidence rate 89.38%, and the total coincidence rate 86.34%. 30 serum samples showed inconsistent, and were detected by Indirect Immunofluorescence Assay (IFA). The results showed: compared with IFA the accuracy rate of result detected by NS3 indirect ELISA and CSFV-Ab test kit of IDEXX was 56.67% and 43.33% respectively.
%K CSFV
%K NS3
%K Gene clone
%K Prokaryotic expression
%K Western blotting
%K Indirect ELISA
%K IFA<br>CSFV
%K NS3
%K 基因克隆
%K 原核表达
%K 蛋白纯化
%K 免疫印迹
%K 间接ELISA
%K IFA
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=78024727B5F4EF6AA9CA8E605B5FC464&aid=2374EE939E4A731B19AA4BB3F8FEE906&yid=140ECF96957D60B2&vid=42425781F0B1C26E&iid=CA4FD0336C81A37A&sid=F9069ED435833594&eid=B5A8A26068AC6AD1&journal_id=0253-2654&journal_name=微生物学通报&referenced_num=1&reference_num=0