基础医学与临床2013，Vol.33Issue(3)：350-355,6.
单纯疱疹病毒1型糖蛋白D主要抗原表位区的表达、纯化及鉴定
Prokaryotic expression and purification of major epitopes domain of herpes simplex virus type 1 glycoprotein D
刘红 1刘宾 2和骁 1赵晓涛1
作者信息
- 1. 北京大学人民医院检验科,北京100044
- 2. 北京大学工学院生物医学工程系,北京100871
- 折叠
摘要
Abstract
Objective To express the major epitope domain of herpes simplex virus type 1 (HSV-1) glycoprotein D (gD) in prokaryotic expression system. The fusion protein was purified and its immunogenicity was studied. Methods The potential epitopes of HSV-1 gD were predicted using the software Protean of Lasergen 7. 0. Then gD major epitopes domain (gD MED) was chosen based on the prediction results, whose cDNA sequence was synthesized and inserted into the prokaryotic expression vector pET-GST. The recombinant gD MED protein was expressed in E. coli BL21(DE3) pLysS with the induction of Isopropyl-β-D-1-Thiogalactopyrano-side (IPTG) and purified with GST · Bind purification kit. The immunological activity was analyzed by western blot. Results HSV-1 gD MED was located at 266 ~ 394 region according to prediction results, whose cDNA sequence was synthesized and its prokaryotic expression vector pET-GST-gD MED was constructed successfully. The fusion gD MED protein could be expressed in soluble form, and its molecular weight (approximately 45 ku) was very close to predicted value. After purification, the purity quotient of the recombinant protein was over 95%. Western blot analysis revealed that the recombinant protein had immunological activity. Conclusions HSV-1 gD MED fusion protein with immunological activity was expressed and purified, which would be helpful to prepare HSV immunologic diagnosis kit and genetically engineering vaccine.关键词
单纯疱疹病毒1型/糖蛋白D/主要抗原表位区/原核表达Key words
herpes simplex virus type 1/ glycoprotein D/ major epitopes domain/ prokaryotic expression分类
医药卫生引用本文复制引用
刘红,刘宾,和骁,赵晓涛..单纯疱疹病毒1型糖蛋白D主要抗原表位区的表达、纯化及鉴定[J].基础医学与临床,2013,33(3):350-355,6.
