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目的 在痘苗系统中表达人CD19基因 ,为研究CD19分子的结构、功能及研制抗CD19的单克隆抗体(mAb)打下基础。方法 用PCR技术扩增人CD19全长基因 ,并重组入克隆载体 pGEM T ,进行序列测定。将CD19全长基因克隆入痘苗表达载体pJSA1175中 ,用脂质体介导的方法 ,与野生病毒共转染TK - 143细胞。运用蓝斑筛选获得含人CD19全长基因的重组痘苗病毒 ,并用此重组病毒感染TK - 143细胞 ,以APAAP法检测人CD19基因在真核细胞中的表达。结果 含人CD19基因的重组痘苗病毒可表达在TK - 143真核细胞的表面。结论 人CD19全长基因在痘苗系统中成功地得到表达
Objective To express human CD19 gene in vaccinia system and lay the foundation for the study of the structure and function of CD19 molecule and the development of anti-CD19 monoclonal antibody (mAb). Methods The full-length human CD19 gene was amplified by PCR and cloned into the cloning vector pGEM T for sequencing. The CD19 full - length gene was cloned into the vaccinia expression vector pJSA1175 and cotransfected into TK - 143 cells with wild - type virus by liposome - mediated method. The recombinant vaccinia virus containing human CD19 full - length gene was obtained by locust screening. The recombinant virus was used to infect TK - 143 cells. APAAP method was used to detect the expression of human CD19 gene in eukaryotic cells. Results Recombinant vaccinia virus containing human CD19 gene was expressed on the surface of TK - 143 eukaryotic cells. Conclusion The full-length human CD19 gene was successfully expressed in the vaccinia system