【摘要】 人小肠三叶因子的cDNA ,克隆到原核表达载体 pGEX 4T 1中 ,经IPTG诱导后 ,hITF以GST hITF融合蛋白的形式表达 ,融合蛋白占细菌总蛋白的 15%左右。经过Glutathione Sepharose4B亲和柱和Sephacry1S 10 0分子筛层析纯化了hITF。SDS PAGE和氨基酸分析证明得到了hITF纯品。以天然hITF为抗原免疫青紫蓝兔制备了抗血清 ,用纯化的IgG建立了hITF的酶联免疫吸附检测 (ELISA)方法 ,给出了标准工作曲线。应用于纯化过程中样品峰的确立。更多还原

【Abstract】 An efficient prokaryotic expression hITF system was established in \%E.coli\% with pGTF through PCR amplification. The amount of fusion protein GST\|hITF was about 15% of the total cellular proteins.The molecular weight of rhITF was 6?kD which was correspondent with that expected.Using hITF as antigen, the antiserum was produced in rabbit.Then enzyme\|linked immunosorbent assay (ELISA) was developed for detection and quantitative analysis of human intestinal trefoil factor, and the standard curves were supplied.更多还原