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首页> 外文期刊>Journal of Huazhong University of Science and Technology >Construction of Eukaryotic Expression Plasmid of Human PRX3 and Its Expression in HEK-293FT Cells
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Construction of Eukaryotic Expression Plasmid of Human PRX3 and Its Expression in HEK-293FT Cells

机译:人PRX3真核表达质粒的构建及其在HEK-293FT细胞中的表达

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To construct the eukaryotic expression plasmid of human PRX3 and measure its expression in the HEK-293FT cells, the full-length coding region of human PRX3 was cloned by PCR and inserted into the eukaryotic expression vector pcDNA4-Xpress (A). HEK-293FT cells were transiently transfected with the recombinant plasmid. Western blot and immuofluorescence were used to detect the expression of the fusion protein. In the experiment, restriction analysis identified the construction of the recombinant plasmid and the inserted sequence was identical with that published on GenBank. Western blot and immunofluorescence confirmed the expression of the recombinant protein in transfected HEK-293FT cells. It was concluded that the eukaryotic expression plasmid of human PRX3 was constructed successfully and the recombinant could be expressed efficiently in HEK-293FT cells, which provides a sound basis for the further study on human PRX3.
机译:为了构建人PRX3的真核表达质粒并测定其在HEK-293FT细胞中的表达,通过PCR克隆人PRX3的全长编码区,并将其插入到真核表达载体pcDNA4-Xpress(A)中。用重组质粒瞬时转染HEK-293FT细胞。用蛋白质印迹和免疫荧光检测融合蛋白的表达。在实验中,限制性酶切分析鉴定了重组质粒的构建,并且插入的序列与GenBank上公布的序列相同。 Western印迹和免疫荧光证实了重组蛋白在转染的HEK-293FT细胞中的表达。结论成功构建了人PRX3的真核表达质粒,可在HEK-293FT细胞中有效表达,为进一步研究人PRX3提供了良好的基础。

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