%0 Journal Article
%T Influence of Epitope A Modification and N-linked Glycosylated Site Mutation of PRRSV NJ-a Strain ORF5 Gene on the Ability to Induce Neutralizing Antibodies and T Cell Proliferation Response<br>PRRSV NJ-a株ORF5基因A表位的修饰与糖基化位点的突变对其DNA疫苗免疫效力的影响
%A ZHENG Qi-Sheng
%A LI Peng
%A BI Zhi-Xiang
%A NIU Ming-Fu
%A CAO Rui-Bing
%A ZHOU Bin
%A CHEN De-Sheng
%A CHEN Pu-Yan
%A <br>郑其升
%A 李鹏
%A 毕志香
%A 牛明福
%A 曹瑞兵
%A 周斌
%A 陈德胜
%A 陈溥言
%J 生物工程学报
%D 2007
%I 
%X To enhance the DNA immunogencity of PRRSV ORF5 gene, CpG sequence and the universal helper T cell antigen epitope (PADRE) sequence were inserted between the decoy epitope and the neutralizing epitope. At the same time, site-mutations were introduced at N33 and N51 to diminish the coverage effect to epitope B from the polysaccharides. Subsequently, the modified ORF5 gene (MORF5) and PRRSV ORF6 gene were cloned into the eukaryotic expression vector pcDNA3.0 under the control of two CMV promoters, respectively. With indirect immunofluorescence assay and Western-blot the expression in vitro of the two genes was confirmed, then six-week-old Balb/C mouse were immunized with the modified expression plasmid pcDNA-M5A-6A. The non-modified expression plasmid pcDNA-5A-6A, the blank eukaryotic expression plasmid pcDNA3.0, living attenuated vaccine and inactivated vaccine were used as controls. The PRRSV specific neutralizing antibodies and the T cell proliferation response were elevated with virus neutralization assay and MTT method. Results indicate that the modified plasmid pcDNA-M5A-6A can elicit not only higher titer of neutralizing antibodies in a rapid time, but also more vigorous T cell proliferation response compared with the non-modified plasmid pcDNA-5A-6A and commercial vaccines, indicating that DNA vaccine pcDNA-M5A-6A maybe a promising candidate for PRRS prevention.
%K Porcine Reproductive and Respiratory Syndrome Virus (PRRSV)
%K ORF5 gene
%K Gene Modification
%K DNA Vaccine<br>猪繁殖与呼吸综合征病毒
%K ORF5基因
%K 基因改造
%K DNA疫苗
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=D52DEE96665155D0&yid=A732AF04DDA03BB3&vid=EA389574707BDED3&iid=CA4FD0336C81A37A&sid=27746BCEEE58E9DC&eid=7C3A4C1EE6A45749&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=6&reference_num=13