融合蛋白IFN-IgGFc在乳酸乳杆菌中的表达及其生物学活性研究  

Expression and Bioactivities Study on IFN-IgG Fc Fusion Protein in Lactobacillus casei

作　　者：高绪文[1] 陈辉[2] 贾润清[3] 姚立红[3] 马长伟[1] 张智清[3] 

机构地区：[1]中国农业大学食品科学与营养工程学院,北京100083 [2]首都医科大学,北京100069 [3]中国疾病预防控制中心病毒病预防控制所,北京100052

出　　处：《食品科学》2007年第1期205-208,共4页Food Science

摘　　要：用乳酸菌表达有药用价值的生物学活性多肽,是当前国际上的研究热点之一,对开发新的功能性食品具有广阔的前景。本文应用PCR方法将人干扰素IFN-α-2b基因与IgGFc基因连接,并引入一段柔性连接肽,构建了融合蛋白基因IFN-IgGFc,将融合蛋白基因克隆到原核表达载体pSC111AE中,转化乳酸乳杆菌ATCC393进行诱导表达及鉴定,并研究了融合蛋白的生物学活性。结果表明,通过Western-Blot方法在表达上清液检测出干扰素融合蛋白,ELISA测定表明两段多肽能够保持各自的构象,基因工程乳酸菌表达的上清液可以诱导WISH细胞产生明显的抗VSV病毒的活性,其活性为1.71×103IU/ml。本文首次构建并在大肠杆菌和乳酸菌表达了IFN-IgGFc融合蛋白,获得了能产生人干扰素的乳酸菌株,为基因工程乳酸菌及相关药物的开发研究奠定了基础。Expressing bioactive polypeptide by using gene modified LAB is one of the most popular researches in the world. It has a good potential to develop new functional food. IFN-α-2b and IgG Fc gene were PCR amplified and linked by a flexible hinge to code for a fusion protein IFN-IgG Fc. The fusion gene was cloned to pSC111 AE vector and electro-transformed into Lactobacillus casei ATCC393. We studied the biological activities of the expression product in vitro. The results showed that we can detect the fusion protein in the supematant by Western-Blot. The results of ELISA showed that there is no effect on conformation and biological activity between the two proteins. We found that supematant of recombinant LAB can introduce the antiviral activity in WISH cells and the anti-viral activity of the product is about 1.71×10^3IU/ml. The study laid a foundation for further use of modified LAB in the domains of food technology and drug development.

关 键 词：乳酸菌 干扰素 融合蛋白 抗病毒 

分 类 号：Q786[生物学—分子生物学]