鼠抗人纤维蛋白单链抗体-低相对分子质量单链尿激酶融合基因真核分泌表达载体的构建

Construction of eukaryotic expression vectors for mouse anti-human fibrin scFv and lowmolecular weight single-chain urokinase fusion gene

目的 构建鼠抗人纤维蛋白单链抗体－低相对单链尿激酶融合基因真核分泌表达载体。方法 应用重组ＤＮＡ技 术，将人工合成的尿激酶原信号肽基因与鼠抗人纤维蛋白单链抗体－低相对分子质量单链尿激酶融合基因连接，并保 证在同一阅读框架，然后分别插科到pcＤＮＡ３和ＰＭＪＫ３两种真核表达载体中。结果构建成可以在真核细胞中分泌表 达鼠抗人纤维蛋白单链抗体－低相对分子质量单链尿激酶融合蛋白的重组质粒pcＤＮＡ３－Ｄ１和ｐＭＪＫ３－Ｄ１。结论为建 立稳定分泌表达鼠抗人纤维蛋白单链抗体－低相对分子质量单链尿激酶融合蛋白的细胞工程株奠定了基础。

Objective To construct the eukaryotic expression vector for the fusion gene comprising the genes encoding mouse anti-human cross-linked fibrin single-chain fragment variable (scFv) antibody and low-molecular weight single-chain urokinase. Methods The single peptide of recombinant human ipro-urokinase was ligated with the target fusion gene, and the same reading frame was guaranteed. The ligated compound was subsequently incorporated respectively into pcDNA3 and pMJK expression vectors by DNA recombination technique. Results and Conclusion The desired recombinant plasmids pcDNA3 and pMJK3 containing the signal peptide of recombinant human ipro-urokinase gene and the fusion gene were constructed,which lays the foundation for establishing of a cell line that may secrete the fusion protein.