单纯疱疹病毒1型糖蛋白B基因疫苗的构建

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-1gpB)基因作为基因疫苗的可能性。方法：利用PCR技术从HSV-1 SM44毒株基因组中扩增出编码HSV-1gpB去除N端部分信号肽序列(39 bp)的基因片断（2 673 bp），定向插入真核表达质粒pcDNA3载体中,构建出重组真核表达质粒pcDNA-gpB,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pcDNA-gpB，电泳可见两条带，分别为目的基因（2 700 bp）和线性质粒pcDNA3（5 400 bp）; 以重组质粒pcDNA-gpB为模板进行PCR扩增，在2 700 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GenBank中登录的HSV-1 F株gpB基因序列比较，同源性达99.5%。 结论：利用PCR技术从HSV-1 SM44株基因组中扩增出编码HSV-1gpB去除N端部分信号肽序列（39 bp）的基因片断（2 673 bp）， 成功地构建了HSV-1基因疫苗pcDNA-gpB。

Construction of herpes simplex virus type 1 glycoprotein B DNA vaccine

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.