摘要
为研制具有较强细胞免疫作用的单纯疱疹病毒Ⅱ型(HSV-2)疫苗,将HSV-2糖蛋白D(gD)序列与糖蛋白B(gB)上两种CTL表位序列连接,人工合成重组基因gD-gBCTL,通过PCR扩增,将其克隆后连接至原核表达载体pET-22b(+),构建重组基因表达质粒pET-22b(+)-gD-gBCTL,转化E.coliBL21(DE3),IPTG诱导表达。对表达产物进行SDS-PAGE分析和Western blot鉴定,His TrapTMHP柱纯化目的蛋白并免疫Balb/c小鼠,间接ELISA法检测血清抗体效价,结果显示重组蛋白在大肠埃希菌中得到了高效表达,该重组蛋白具有较好的免疫原性。
Herpes simplex virus type Ⅱ（HSV-2） glycoprotein D（gD） is one of the most important viral immunogen,glycoprotein B（gB） CTL epitope could stimulate the formation of cytotoxic CD8＋T cells.The sequences of HSV-2 gD and gB CTL epitopes were linked artificially,and the recombinant gene gD-gB CTL constructed.The new gene was successfully expressed in prokaryotic expression system.The expressed protein was analyzed by SDS-PAGE and Western blot.The recombinant protein was purified to immunize Balb/c mice,while seral specific antibody titers were measured by indirect ELISA.The results indicated that the recombinant protein has good immunogenicity.
出处
《动物医学进展》
CSCD
北大核心
2011年第6期19-23,共5页
Progress In Veterinary Medicine
