一种新型单纯疱疹病毒II型DNA疫苗的构建及其初步研究  

Primary research of a new herpes simplex virus-2 DNA vaccine

作　　者：何方[1] 洪艳[1] 陈勇[1] 

机构地区：[1]浙江省医学科学院生物工程所,杭州310013

出　　处：《中国卫生检验杂志》2008年第6期997-999,共3页Chinese Journal of Health Laboratory Technology

基　　金：浙江省科技厅资助项目(2005F13022)

摘　　要：目的:构建新型的含单纯疱疹病毒II型糖蛋白D基因(gD)的卡那霉素抗性真核表达质粒pgD,并探讨其作为DNA疫苗的可能性。方法:通过PCR扩增、酶切、连接及克隆筛选从质粒pET-28a(+)和质粒pcDNA3获得新型真核表达质粒载体pcDNAkan。再通过PCR从重组真核表达质粒pcDNA3-gD上扩增出HSV-2糖蛋白D全基因序列(gD),最后克隆重组获得含gD的重组质粒pgD。pgD作为DNA疫苗免疫小鼠,观察其对小鼠的免疫效果及保护作用。结果:pgD免疫小鼠后能产生特异性抗体,与对照组相比较具有显著性差异(P<0.01),在HSV-2病毒致死性攻毒实验中,pgD组存活率达75%。结论:重组质粒pgD能诱导小鼠产生特异性免疫应答,可为HSV的DNA疫苗研制提供科学依据。Objective： To construct a new eukaryotic expression plasmid pgD encoding gene kan^＋ and herpes simplex virus - 2 （ HSV - 2） full length glycoprotein D （ gD ） to evaluate the protective efficacy of pgD as HSV - 2 DNA vaccine. Methods： Acquired a new eukaryotic expression plasmid vector pcDNAkan from plasmid pET- 28a（ ＋ ） and pcDNA3 by PCR and T4 DNA ligase. Gene encoding full length HSV - 2 gD was amplified from plasmid pcDNA3 - gD, and cloned into multiple cloning sites of the eukaryotic expression plasmid vector pcDNAkan, resulted in recombined plasmid pgD. PgD inoculated to mice by bilateral intramuscular injection into the rear legs, and its protective efficacy was observed. Results ：The mice inoculated with pgD could produce specific antibodies and 75% of the mice survived after lethal challenge with HSV- 2. Conclusion：pgD can induce specific immune response. The present study provides a scientific basis for the HSV DNA vaccine research.

关 键 词：单纯疱疹病毒 DNA疫苗 PCR 糖蛋白D 

分 类 号：R37[医药卫生—病原生物学]