[关键词]
[摘要]
构建含D 氨基酸氧化酶 (DAAO)基因的重组逆转录病毒载体 ,初步观察DAAO基因的功能。方法 :利用重组DNA技术将DAAOcDNA亚克隆至逆转录病毒载体 (pLDAAOSN)中 ,以磷酸钙沉淀法介导转染包装细胞ΦXNA ,用NIH3T3细胞测定病毒滴度 ,将重组逆转录病毒感染K5 6 2白血病细胞 ,G418筛选出抗性克隆 ,命名为KDAAO。PCR、原位杂交分析外源基因整合和表达 ,观察 5 0mm/LD 丙氨酸对KDAAO杀伤作用。结果 :经酶切鉴定和DNA测序证明重组逆转录病毒载体中含有完整的DAAO基因。包装细胞产生了高滴度病毒 (5 .2× 10 6CFU/ml)。PCR、原位杂交分析证明DAAO基因已整合至KDAAO基因组中 ,并在mRNA水平表达。初步观察到D 丙氨酸能明显杀伤KDAAO。结论 :重组逆转录病毒载体pLDAAOSN可以进一步用于肿瘤的基因治疗研究
[Key word]
[Abstract]
Objective: To research influence of consecutive Immunization on cellular and humoral immunity in mice. Methods: We evaluated a consecutive immunization strategy priming with recombinant fowlpox virus vUTALG and boosting with nucleic acid vaccine plasmid pcDNAG encoding HIV-1 capsid protein Gag. Results: In immunized mice, the number of CD4 cells from splenic lymphocytes increased singnificantly and the proliferation response of splenocytes to Con A, LPS elevated markedly and HIV-1-specific antibody response could be induced. Conclusion: Consecutive immunization could increase cellular and humoral immunity response in mice.
[中图分类号]
R392 R394
[基金项目]
本课题由国家自然科学基金重点项目(39870710)资助
