【摘 要】
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Objective To construct eukaryotic expression plasmid of recombinant human MED12-Fc using the pIRES2-EGFP-Fc vector.Methods According to the information in GenBank,the sequence of exons 2 to 4 of human
【机 构】
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Department of general surgery,Taizhou people's Hospital,Taizhou Jiangsu 225300,China
【出 处】
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中华医学会第八次全国中青年检验医学学术会议
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Objective To construct eukaryotic expression plasmid of recombinant human MED12-Fc using the pIRES2-EGFP-Fc vector.Methods According to the information in GenBank,the sequence of exons 2 to 4 of human MED12 gene was amplified from the genome.The target fragment was obtained by OVERLAP-PCR splicing and inserted into the pIRES2-EGFP-Fc vector.The expression plasmid was transfected into CHO cells by electroporation.The purified product was analyzed by SDS-PAGE.Results DNA sequencing and expression of transient transfection indicated exons 2 to 4 of MED12 gene sequence was correct and MED12-Fc had been constructed successfully.The positive clone with high expressing fusion protein was got by electroporation.
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