摘要
目的构建HSV 1型SM44株糖蛋白D(gD)基因的真核表达载体 ,并用此重组质粒直接免疫小鼠 ,探讨HSV 1gD基因作为基因疫苗的可能性。方法从HSV 1基因组中扩增 gD的全编码基因 ,克隆入载体 pUC19中 ,测序鉴定后转入真核表达载体 pcDNA3.1( +)。所得重组质粒pcDNA gD以电穿孔法转染CHO细胞 ,并以荧光染色法鉴定表达效果。用 pcDNA gD免疫小鼠 ,ELISA法检测基因免疫的应答效果。结果成功地构建了可在真核细胞中有效表达的HSV 1gD真核表达载体 ,用其直接免疫小鼠可引起较高水平的特异性抗体应答。结论 gD的真核表达载体有可能作为HSV 1的基因疫苗 ,为进一步研究基于 gD的表位生物学和表位疫苗奠定了基础。
Aim To construct eukaryotic expression vector bearing HSV-1 glycoprotein D(gD) gene and to evaluate the possibility of designing HSV-1 gene vaccine with HSV gD gene. Methods The HSV 1 gD gene was amplified from the HSV 1 genome DNA and cloned into eukaryotic vector pcDNA3.1(+). The recombinant vector pcDNA gD was transfected into CHO cells by electroperforance method. Fluorescent staining and ELISA were used to test the expression of gD in CHO cells and response of mice to pcDNA gD. Results The eukaryotic expression vector containing HSV 1 gD gene was successfully constructed .The recombinant vector could induce higher specific immunoreaction in the immunized mice. Conclusion The recombinant vector pcDNA gD is valuable to designing HSV gene vaccine and studying epitope biology and epitope vaccine of HSV 1.
出处
《细胞与分子免疫学杂志》
CSCD
2000年第4期308-310,共3页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金资助 !No.39770040
关键词
HSV-1
糖蛋白D
真核表达
基因免疫
载体构建
herpes simplex virus 1
glycoprotein D
eu
karyotic expression
gene immunization
