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.

Key words: intestinal trefoil factor, enzyme-linked immunosorbent assay (ELISA), clone and expression

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-Sepharose 4B亲和柱和Sephacry1 S 100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。

关键词: 小肠三叶因子, 酶联免疫吸附检测, 克隆表达