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目的构建人IgG3上游铰链区/p53四价功能域融合基因，并进行空间构象的分析。方法利用递归PCR法，扩增人IgG3上游铰链区/p53四价功能域融合基因，并在融合基因5′端和3′端引入Sal I与Hind Ⅲ酶切位点。将融合基因克隆入pUC19载体中，经酶切鉴定及序列测定证实后，利用计算机软件进行空间构象分析。结果人IgG3上游铰链区/p53四价功能域融合基因，经酶切鉴定及测序证实，其大小及核苷酸序列正确，与设计完全一致。计算机软件分析显示，融合基因表达产物可自动装配成四聚体，并具有4条柔性好的长多肽，可确保与其融合的其他基因片段空间构象的独立性。结论人IgG3上游铰链区/p53四价功能域融合基因的成功构建，为进一步研制多价抗体奠定了基础。
Objective To construct the tetravalent fusion protein of human IgG3 upstream hinge region / p53 and analyze the spatial conformation. Methods The reversion PCR method was used to amplify the tetravalent fusion protein of human IgG3 hinge region / p53 and to introduce Sal I and Hind Ⅲ restriction sites at the 5 ’and 3’ ends of the fusion gene. The fusion gene was cloned into pUC19 vector, and confirmed by restriction enzyme digestion and sequence analysis, the spatial conformation was analyzed by computer software. Results The human IgG3 upstream hinge region / p53 tetravalent fusion gene was verified by restriction enzyme digestion and sequencing. The size and nucleotide sequence were correct and consistent with the design. Computer software analysis showed that the fusion gene expression products could be automatically assembled into tetramers and had 4 flexible long peptides to ensure the spatial conformational independence of other gene fragments fused with them. Conclusion The successful construction of the fusion region of the upstream hinge region of human IgG3 / p53 gene provides the basis for further development of multivalent antibody.