单纯疱疹病毒1型糖蛋白B基因疫苗的构建

doi:吉林大学新技术新疗法基金资助课题；长春市

单纯疱疹病毒1型糖蛋白B基因疫苗的构建

孟祥俊¹，贺冰²，冯非²，李光源²，毕军³

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

收稿日期:2004-03-22 修回日期:1900-01-01 出版日期:2004-09-28 发布日期:2004-09-28

Construction of herpes simplex virus type 1glycoprotein B DNA vaccineMENG

Xiang-jun¹, HE Bing², FENG Fei²,LI Guang-yuan²,BI Jun³

1. Department of Ophthalmology, Second Hospital, Jilin University， Changchun 130041, China;2. Department of Ophthalmology, First Hospital, Jilin University， Changchun 130021,China;3. Department of Internal Medicine, Huinan Hospital of Traditional Chinese Medicine, Huinan 135100, China

Received:2004-03-22 Revised:1900-01-01 Online:2004-09-28 Published:2004-09-28

摘要/Abstract

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-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。

关键词: 糖蛋白B, DNA疫苗

Abstract: 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.

Key words: glycoprotein B, DNA vaccines

中图分类号:

孟祥俊，贺冰，冯非，李光源，毕军. 单纯疱疹病毒1型糖蛋白B基因疫苗的构建[J]. J4, 2004, 30(5): 717-720.

Xiang-jun, HE Bing, FENG Fei,LI Guang-yuan,BI Jun. Construction of herpes simplex virus type 1glycoprotein B DNA vaccineMENG[J]. J4, 2004, 30(5): 717-720.

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