单纯疱疹病毒1型糖蛋白B基因疫苗的构建  

Construction of herpes simplex virus type 1 glycoprotein B DNA vaccine

作　　者：孟祥俊[1] 贺冰[2] 冯非[2] 李光源[2] 毕军[3] 

机构地区：[1]吉林大学第二医院眼科,吉林长春130041 [2]吉林大学第一医院眼科,吉林长春130021 [3]吉林省辉南县中医院内科,吉林辉南135100

出　　处：《吉林大学学报（医学版）》2004年第5期717-720,共4页Journal of Jilin University：Medicine Edition

基　　金：吉林大学新技术新疗法基金资助课题 (2 0 0 10 4 ) ;长春市科技发展计划项目资助课题 (0 3- 170 5 0 9)

摘　　要：目的 :构建用于治疗和预防单纯疱疹病毒性角膜炎的核酸疫苗 ,探讨单纯疱疹病毒 1型糖蛋白 B(HSV-1gp B)基因作为基因疫苗的可能性。方法 :利用 PCR技术从 HSV- 1SM4 4毒株基因组中扩增出编码 HSV- 1gp B去除 N端部分信号肽序列 (39bp)的基因片断 (2 6 73bp) ,定向插入真核表达质粒 pc DNA3载体中 ,构建出重组真核表达质粒 pc DNA- gp B,并对其进行酶切分析、PCR鉴定及测序鉴定。结果 :双酶切重组质粒 pc DNA- gp B,电泳可见两条带 ,分别为目的基因 (2 70 0 bp)和线性质粒 pc DNA3(5 4 0 0 bp) ;以重组质粒 pc DNA- gp B为模板进行 PCR扩增 ,在 2 70 0 bp位置扩增出特异的产物 ;测序结果表明 ,克隆基因插入方向正确 ,与 Gen Bank中登录的 HSV- 1F株 gp B基因序列比较 ,同源性达 99.5 %。结论 :利用 PCR技术从 HSV- 1SM4 4株基因组中扩增出编码 HSV- 1gp B去除 N端部分信号肽序列 (39bp)的基因片断 (2 6 73bp) ,成功地构建了 HSV- 1基因疫苗pc DNA- gp B。Objective To construct the DNA vaccine that would be used to prevent and treat herpes simplex keratitis,and to evaluate the possibility of designing HSV-1 gene vaccine with HSV gpB gene. Methods The part encoding sequence of the glycoprotein B (gpB) was amplified from HSV-1 SM44 DNA genome by polymerase chain reaction (PCR), and then was directionally cloned into eukaryotic expression vector pcDNA3, the recombinant vector pcDNA-gpB was confirmed by the restriction endonuclease analysis, PCR and sequence analysis. Results Double-enzyme digestion and analysis of the recombinant vector pcDNA-gpB showed two bands, one was HSV-1gpB gene(2 700 bp),the other was linear pcDNA3 (5 400 bp);HSV-1gpB gene was cloned by pcDNA-gpB as template; sequence analysis showed orientation was right and the rate of homology was 99.5% compared with GenBank. Conclusion The part encoding sequence of the glycoprotein B is cloned and the recombinant vector pcDNA-gpB is constructed successfully.

关 键 词：单纯疱疹病毒1型 糖蛋白B DNA疫苗 

分 类 号：Q78[生物学—分子生物学]