Cloning and Expression of an Immunodominant Fragment of Glycoprotein G of HSV-2 Using Directional TOPO Vector in E.coli
- VernacularTitle:TOPO定向克隆和表达Ⅱ型单纯疱疹病毒gG基因的免疫优势表位片段
- Author:
Shuqi WANG
;
Yueping YIN
;
Yanhua YU
;
Weihong LAI
;
Mingjun JIANG
;
Qiang CHEN
;
Xiangsheng CHEN
- Publication Type:Journal Article
- Keywords:
Herpesvirus 2, human;
Genes, viral;
Gene expression
- From:
Chinese Journal of Dermatology
1994;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To clone and express immunodominant fragment of glycoprotein G of HSV-2 (FgG-2). Methods The target gene was amplified by polymerase chain reaction (PCR). The PCR products were ligated into directional TOPO expression vector. After identification, the recombinant expression vector was transferred into BL21 StarTM cell for expression. Finally, recombinant protein of FgG-2 (rFgG-2) was detected by Western Blot (WB). Results A 616 bp DNA fragment was obtained with PCR and then confirmed in recombinant vector by PCR and sequencing, bearing 99.5% consistent sequence with target gene. Highest recombinant protein production was obtained at the time point of 3 hours. Expression of target protein was confirmed by WB with anti-gG monoclonal antibody. Conclusions The immunodominant fragment of gG-2 has been successfully cloned and expressed in E.coli, which might be used for the development of serum diagnostics assay kits for HSV-2 infection.
