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痘苗病毒的基因组庞大，结构复杂而特殊，不可能将外源基因直接插入它的基因组，必须利用一种特殊的痘苗病毒质粒，才能构建成功重组痘苗病毒．在分析了痘苗病毒质粒ｐＪ１２０〔含有我国天花疫苗－痘苗病毒天坛株７６１的启动子和胸苷激酶（ｔｈｙｍｉｄｉｎｅｋｉｎａｓｅ，简称ＴＫ基因），及含有人癌胚抗原（ｃａｒｃｉｎｏｅｍｂｒｙｎｉｃａｎｔｉｇｅｎ，简称ＣＥＡ）ｃＤＮＡ全序列的质粒ｐ９１０２３Ｂ－ｃｅａ－１７结构的基础上，设计出三步法构建了重组疫苗病毒质粒ｐＪ－ＣＥＡ．经酶切及ＰＣＲ鉴定ｐＪ－ＣＥＡ中ＣＥＡｃＤ－ＮＡ的存在，进一步用同源重组方法构建了表达人ＣＥＡ的重组痘苗病毒，并以人体成纤维细胞作为宿主细胞，对ＣＥＡ－重组痘苗病毒进行了大量培养．再次证实痘苗病毒是良好的真核表达载体，可以高效而准确地表达细胞膜糖蛋白ＣＥＡ．
Vaccinia virus genome is large, complex and special structure, it is impossible to insert foreign genes directly into its genome, you must use a special vaccinia virus plasmid, in order to build a successful recombinant vaccinia virus. After analyzing the vaccinia virus plasmid pJ120 [including the promoter of the smallpox-vaccinia virus Tiantan strain 761 and the thymidine kinase (TK gene), and the plasmid containing the complete sequence of human carcinoembryonic antigen (CEA) cDNA, p91023B-cea-17 structure based on the design of a three-step method to construct a recombinant vaccine virus plasmid pJ-CEA. CEAcD-NA in pJ-CEA was identified by restriction enzyme digestion and PCR. Recombinant vaccinia virus expressing human CEA was further constructed by homologous recombination. Human fibroblasts were used as host cells for CEA-recombinant vaccinia virus Extensive training. Again confirmed that vaccinia virus is a good eukaryotic expression vector, can be highly efficient and accurate expression of cell membrane glycoprotein CEA.