通用型转铁蛋白融合表达载体的构建及应用价值

Construction and Application Value of Universal Transferrin Fusion Expression Vector

构建通用型转铁蛋白融合表达载体,利用PCR方法扩增编码人转铁蛋白N端半分子的基因片段,通过酶切、连接、转化等分子克隆方法构建通用型转铁蛋白融合表达载体。PCR扩增了一个长约1.1 kb的包含ScaI酶切位点的基因片段,插入pPICZα的PmlI和XbaI酶切位点,转化后进行菌液PCR鉴定,成功获得重组子pPICZα-TfN,测序结果表明载体构建成功,重组质粒pPICZα-TfN能被ScaI酶切。本研究成功构建通用型转铁蛋白融合表达载体,构建的载体可以用于转铁蛋白融合表达载体的构建。

To construct the universal transferrin fusion expression vector,PCR was employed to amplify the N-terminal half-molecular of human transferrin gene.Molecular cloning methods,including digestion,ligation,transformation were used to construct the universal transferrin fusion expression vector.A 1.1 kb DNA fragment containing the Sca I site was amplified and it was inserted into Pml I and Xba I sites of pPICZα.After transformation and PCR identification,recombinant plasmid pPICZα-TfN was obtained successfully.Sequencing analysis indicated that the open reading frame was correctly.The plasmid pPICZα-TfN can be linearized by Sca I.The universal transferrin fusion expression vector was constructed successfully in this study.