%0 Journal Article
%T MOLECLONING AND EXPRKSSION OF E2 GENE OF THE CHINKSE CLASSICAL SWINE FEVER VIRUS(SHIMEN STRAIN) AND PRELIMINARY STUDIES OF ITS DNA VACCINE<br>猪瘟病毒E2(gp55)基因的克隆表达及其DNA疫苗的初步研究
%A Zhou Pengcheng
%A Lu Yu
%A Chen Jianguo
%A Zhai Zhonghe
%A Ding Mingxiao
%A <br>周鹏程
%A 陆宇
%A 陈建国
%A 翟中和
%A 丁明孝
%J 微生物学报
%D 2000
%I 
%X A 1 1bp fragment of E2 gene of chinese classical swine fever virus(CSFV)Shimen strain, a standard virulent strain, was amplified by RT PCR from total RNA of cell cultures infected by CSFV, and cloned into pGEM T vector. the nucleotide sequence of this fragment was sequenced by Sanger's method and the amino acid sequence was deduced. Compared with the corresponding region of Alfort, Brescia and C strain of CSFV, the nucleotide sequence homology is 84 7%, 92 6% and 95 2% respectively, and the amino acid sequence 89 4%, 92 6% and 94 6%, respectively. we subcloned 1 1bp of E2 gene cDNA into baculovirus transfer vector and successfully constructed two recombinant baculoviruses expressing GST E2 and GST GFP E2 fusion protein respectively by homologous recombination in sf 9 cell. Furthermore, we also constructed recombinant eukaryotic expression vector pcE2 containing E2 gene in frame and transfected COS 7 cell by lipofectamine, the indirect immunofluorescence assay (IFA) showed that the expressed E2 protein can be recognized by E2 specific monoclnal antibody. the pcE2 DNA was directly injected into BALB/c mice intramusclarly(i.m) and the CSFV E2 specific antibodies was measured by enzyme linked immunosorbent assay(ELISA). the ELISA results indicated the E2 specific antibodies was induced in inoculated mice and virus neutralization assays also indicate single inoculations of plasmids expressing CSFV E2 glycoprotein raised neutralizing antibody in BALB/c mice. these results will be beneficial to investigate the possibility of DNA caccine against CSFV.
%K Classical swine fever virus
%K Clone and expression
%K DNA vaccine<br>猪瘟病毒，
%K 克隆表达，
%K DNA疫苗
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A3C6BA55AB623B90FA9104CFFC826F3C&aid=6167A4B0985E93EC&yid=9806D0D4EAA9BED3&vid=1371F55DA51B6E64&iid=38B194292C032A66&sid=B78CD622C1934236&eid=4B1FFFA116F7AE3B&journal_id=0001-6209&journal_name=微生物学报&referenced_num=12&reference_num=13