摘要
目的 :探讨人B细胞淋巴瘤活检组织中肿瘤细胞膜表面免疫球蛋白VH(smIgVH)基因片段能否在动物体内激发特异性抗独特型抗体。方法 :以RT PCR法获得IgVH 基因片段 ,以小鼠单核细胞趋化因子 (MCP 3)基因作为佐剂分子 ,进行重组PCR获得MCP 3和VH 基因片段的融合基因 ,克隆在真核表达载体 pcDNA3.1中 ,构建DNA疫苗质粒 pcDNA/MCP BVH。通过脂质体转染验证该质粒在真核细胞COS 7中的表达。结果 :通过上述方法获得了以活检组织肿瘤细胞mIgVH区基因片段 ;成功地构建了DNA疫苗质粒 pcDNA/MCP BVH。体外瞬时转染实验证明 ,该质粒能够在真核细胞COS 7中正确表达。结论 :成功地构建重组表达质粒 pcDNA/MCPBVH,在体外能够正确表达 。
AIM: To construct a DNA vaccine based on surface Ig V H gene of tumor cells in the human B-cell lymphoma biopsy tissue. METHODS: The V H gene fragment was amplified by RT-PCR using Ig superfamily primers. Also, the murine monocyte chemotactic protein 3(MCP-3) cDNA was cloned. The fusion gene fragment of MCP-3 gene with V H gene was constructed by recombinant PCR and then cloned into the eukaryonic expression vector pcDNA3.1 to construct the DNA vaccine plamid pcDNA3.1/MCP-V H. The vaccine plamid was transiently expressed in the eukaryotic cell line COS-7. RESULTS: The DNA vaccine plasmid was successfully constructed and expressed in COS7 cells in the form of fusion protein MCP-V H. CONCLUSION: The DNA vaccine plamid pcDNA3.1/MCP-V H is constructed and expressed successfully, which plays the foundation for further experimental research in animal model.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2004年第1期79-82,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金资助项目 (No .30 2 71 2 4 5)
