【摘要】 利用PCR技术将人小肠三叶因子(hITF)基因重组入表达载体pGEX-4T-1,构建了融合蛋白GST-hITF的重组表达质粒pGTF,在大肠杆菌中诱导表达。表达的融合蛋白经亲和层析、凝血酶切和凝胶过滤层析得到纯化的hITF蛋白。测定了重组蛋白的氨基酸组成、分子量及其对酸和蛋白酶的抗性。Western印迹表明重组蛋白具有hITF的抗原性,并对大鼠胃溃疡具有明显的预防和保护作用。更多还原

【Abstract】 The cDNA of human intestinal trefoil factor(hITF)was amplified by PCR and cloned into the expression vector pGEX 4T 1,and expressed in E.coli as part of a fusion protein with glutathione S transferase(GST)as the induction of IPTG.Recombinant hITF was purified by affinity chromatography,thrombin digestion and gel filtration.The amino acid composition and molecular mass of purified hITF was determined.hITF was somewhat resistant to acid and proteinase.Western blotting indicated that recombinant protein could react with antibodies against hITF.hITF has precautionary and therapeutic function for hydrochloric acid induced gastric ulcer in rats.更多还原