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目的构建人肿瘤转移抑制基因ＮＭ２３－Ｈ１与真核表达载体ＰｃＤＮＡ３．１／Ｚｅｒｏ的重组体。方法利用分子生物学克隆技术从质粒ｐｕｃ１８中酶切回收９００ｂｐ的人ＮＭ２３－Ｈ１基因片段，与ＰｃＤＮＡ３．１／Ｚｅｒｏ构成重组体，并根据表达载体与克隆基因核苷酸序列中酶切位点分析、鉴定插入片段的正反连接。结果酶切得到９００ｂｐ的ＮＭ２３－Ｈ１基因片段，５．０ｋｂ质粒载体，两者相连构建成重组体ＰｃＤＮＡ．ＮＭ２３，利用ＰｓｔⅠ酶切位点确认ＰｓｔＩ酶切后得到４条带的重组体为正向连接。结论人真核细胞表达质粒ＰｃＤＮＡ．ＮＭ２３成功构建，为卵巢癌基因治疗提供了部分实验基础。
Objective To construct a recombinant human tumor metastasis suppressor gene NM23-H1 and eukaryotic expression vector pcDNA3.1 / Zero. Methods The molecular weight of human bp23 (bp) of human NM23-H1 was digested with restriction endonuclease digestion from plasmid puc18 and cloned into pcDNA3.1 / Zero. According to the analysis of the restriction sites in the nucleotide sequence of the expression vector and cloned gene , Identifying the positive and negative insertions of the insert. Results The 900bp NM23-H1 gene fragment and 5.0kb plasmid vector were obtained by digestion. The two were ligated to construct recombinant PcDNA. NM23, PstI restriction sites were used to confirm PstI digestion to obtain 4 bands of recombinant positive connection. Conclusions Human eukaryotic expression plasmid pcDNA. NM23 was successfully constructed and provided some experimental basis for ovarian cancer gene therapy.