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为获得高质量及充足的Ｆａｓ蛋白，采用ＰＣＲ技术调整Ｆａｓ基因的开放阅读框架，使之与生物素化蛋白基因阅读框架一致；缺失了ＦａｓｃＤＮＡ基因的起始密码子并增加一个大肠杆菌偏性终止密码子，构建ＦａｓｃＤＮＡ和生物素化融合原核表达质粒ＰｉｎＰｏｉｎｔ－Ｆａｓ。将重组质粒转入大肠杆菌ＨＢ１０１，经５００ｍｍｏｌＩＰＴＧ在３７℃条件下诱导４ｈ，ＳＤＳ－ＰＡＧＥ及Ｗｅｓｔｅｒｎ印迹检测融合蛋白在大肠杆菌得以高效表达，表达量为细菌总蛋白的１３．８％。用亲和层析树脂对生物素化融合蛋白进行亲和层析纯化，得到Ｆａｓ重组的蛋白，且表达的Ｆａｓ融合蛋白具有抗体结合活性。此蛋白的表达成功将解决Ｆａｓ膜蛋白不易提取的难题，为深入研究Ｆａｓ提供了良好材料来源
In order to obtain high-quality and sufficient Fas protein, the open reading frame of Fas gene was adjusted by PCR to make it consistent with the biotinylated protein gene reading frame. The start codon of FascDNA gene was deleted and an Escherichia coli biased termination was added Codon to construct Fetal-Fas fusion protein and biotinylated fusion plasmid. The recombinant plasmid was transformed into E.coli HB101 and induced by 500mmol IPTG for 4h at 37 ℃. The fusion protein was highly expressed in E.coli by SDS-PAGE and Western blot. The expressed protein was 13.8% of the total bacterial protein. Affinity chromatography of the biotinylated fusion protein with affinity chromatography resin yielded the recombinant Fas protein, and the expressed Fas fusion protein had antibody binding activity. The successful expression of this protein will solve the difficult problem of Fas membrane protein extraction and provide a good source of material for further study of Fas