中文摘要：

目的：构建pFUSE-hIgG2-Fc2-BRBP1真核表达质粒表达BRBP1-Fc肽体,并验证其与乳腺癌脑转移细胞系（231-BR）的结合特异性。方法：合成带有EcoRⅠ、NcoⅠ酶切位点的脑转移性乳腺癌特异性肽BRBP1片段,利用该双酶切位点将其定向插入pFUSE-hIgG2-Fc2质粒,构建真核表达质粒pFUSE-hIgG2-Fc2-BRBP1。将测序正确的重组质粒转染至293T细胞真核表达BRBP1-Fc肽体,用蛋白质印迹实验验证蛋白表达,酶联免疫吸附试验、细胞免疫荧光等方法证实BRBP1-Fc肽体与231-BR细胞结合特异性。结果：成功构建重组质粒pFUSE-hIgG2-Fc2-BRBP1,蛋白质印迹法检测到BRBP1-Fc肽体的表达,分子质量约为37 k Da,并且证实BRBP1-Fc肽体与231-BR细胞特异性结合。结论：成功构建了真核表达重组质粒pFUSEhIgG2-Fc2-BRBP1,表达了BRBP1-Fc肽体,并鉴定了其与231-BR细胞的结合特异性。

英文摘要：

Objective： To construct eukaryotic expression vector of pFUSE-hIgG2-Fc2-BRBP1 and identify its recombinant protein expression and specific binding with 231-BR cells. Methods： BRBP1 sequence with EcoRⅠ,NcoⅠenzymes was synthesized by gene company and subcloned into pFUSE-hIgG2-Fc2 vector. After the region was sequenced,the plasmid was transfected into 293 T cell line. The expression of the recombinant plasmid in 293 T cells was proved by Western blotting. The specific binding was observed by enzyme linked immunosorbent assay and immunofluorescence method. Results： BRBP1 had been constructed into expression vector pFUSE-hIgG2-Fc2 successfully. The expression of BRBP1-Fc fusion protein with a molecular weight 37 k Da was detected by Western blotting. The binding specificity with 231-BR cell was also identified. Conclusion： We have successfully constructed the recombinant plasmid（ pFUSE-hIgG2-Fc2-BRBP1） which was expressed in 293 T cells and had high binding specificity with 231-BR cell.