作　　者： ; ; ; ; ; ; ;

机构地区： 南京师范大学生命科学学院

出　　处： 《南京师大学报（自然科学版）》 2008年第2期97-100,104,共5页

摘　　要： 对巴斯德毕赤酵母高效分泌表达的猪α-干扰素(PoIFN-α)纯化工艺和纯化后重组蛋白的部分生化特性进行了研究,结果表明:猪α-干扰素(PoIFN-α)发酵液经离心、透析、过滤处理之后,依次利用亲和层析和离子交换层析使目的蛋白得到了纯化.经N端氨基酸测序,Western-blot,对酸、热、巯基乙醇的稳定性和糖基化程度等检测后发现,所表达的猪α-干扰素N端氨基酸序列(前5个)正确,对酸和热基本稳定,二硫键对目的蛋白的活性至关重要,没有发现目的蛋白的糖基化. In this study, we developed the technique for purifying recombinant porcine-α interferon （PolFN-α） that was efficiently expressed by Pichia Pastoris and studied the biochemical characteristics of PolFN-α. Recombinant porcine IFN-α was purified to homogeneity by affinity chromatography and ion-exchange chromatography stepwisely after centrifugation,dialysis and filtration. The results of its biochemical characteristics showed that the first five amino acids of N - terminal of the purified protein was correct. The protein was heat and acid-stable and the dithiolate-bond was crucial to the protein activity. No glycosylation were found on the target protein.

关 键 词： 重组猪 干扰素 亲和层析 离子交换层析 糖基化 生化性质

领　　域： [农业科学] [农业科学] [农业科学]