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从ｐＵＣ１８－３４重组质粒双酶切分离得到编码人ＣＤ＿（３４）抗原的全长ｃＤＮＡ片段，将此基因插入痘苗病毒载体ＳｍａＩ位点，构建了重组质粒ＮＳＡ１１７５－３４。采用Ｌｉｐｏｆｅｃｔｉｎ方法，将质粒ｐＪＳＡ１１７５－３４导入已被野生痘苗病毒感染的ＴＫ￣－１４３细胞，用ＢｕｄＲ和ＬａｃＺ双筛选，获得带有人ＣＤ＿（３４）抗原基因的重组痘苗病毒。经活细胞免疫荧光（ＩＦＡ）和碱性磷酸酶－抗碱性磷酸酶（ＡＰＡＡＰ）检测表明，重组痘苗病毒能够特异地表达人ＣＤ＿（３４）抗原。人ＣＤ＿（３４）抗原基因的表达，为探讨ＣＤ＿（３４）抗原结构和功能以及研制ＣＤ＿（３４）单克隆抗体奠定了基础。
The full length cDNA fragment encoding human CD34 antigen was isolated from the pUC18-34 recombinant plasmid by digestion and inserted into SmaI site of the vaccinia virus vector to construct the recombinant plasmid NSA1175-34. The plasmid pJSA1175-34 was transfected into TK-143 cells infected with wild vaccinia virus by Lipofectin method. The recombinant vaccinia virus with human CD_ (34) antigen gene was obtained by double screening with BudR and LacZ. The results of living cell immunofluorescence assay (IFA) and alkaline phosphatase - alkaline phosphatase (APAAP) assay showed that the recombinant vaccinia virus could specifically express human CD34 antigen. The expression of human CD_ (34) antigen gene laid the foundation for exploring the structure and function of CD_ (34) antigen and the development of CD_ (34) monoclonal antibody.