鸡α干扰素在毕赤酵母中的分泌表达及其抗病毒活性的测定  

Expression of chicken interferon alpha in Pichia pastoris and studies on antiviral effects

作　　者：王秀丽 徐慧琳 刘新文 郭东春[2] 郭伟伟 邹敏 李陆梅 杜元钊 

机构地区：[1]青岛易邦生物工程有限公司,山东青岛266032 [2]中国农业科学院哈尔滨兽医研究所,黑龙江哈尔滨150069

出　　处：《中国兽医科学》2017年第5期626-629,共4页Chinese Veterinary Science

摘　　要：将克隆的鸡α干扰素(ChIFNα)基因亚克隆至酵母表达载体pPICZα-A中,构建重组质粒pPIC-ChIFNα,线性化后电转化整合到毕赤酵母菌株X-33基因组中,得到阳性菌株。经1%甲醇连续诱导4 d,SDS-PAGE检测结果表明鸡α干扰素在酵母中获得了分泌型表达,表达产物分子质量约为18 ku。细胞病变抑制法检测其效价为1×10^(7.69)U/mL;在鸡胚中能有效抑制禽流感病毒H9N2的增殖。针对Ⅰ群4型禽腺病毒YBAV-4株强毒的动物攻毒保护试验结果显示,表达的蛋白具有较好的抗病毒活性。The gene of chicken interferon alpha（ChIFN-a） was was subcloned into the expression vector pPICZα-A. After linearized by Sac I ,the plasmid was transferred into yeast Pichia pastoris strain X-33. The recombinant strain was identified by PCR. The recombinant strain was induced by 1% methanol for 4 days,recombinant ChIFN-a was detected by SDS-PAGE. The assay results showed that the protein was approximately 18 ku in size,and the recombinant protein was secreted effectively in Pichia pastoris. Antiviral activities of the recombinant ChIFN-~ were detected by cell pathological effect inhibition assay. The result showed the titer was about 1 × 10^7.69 U/mL. The expressed protein could inhibit H9N2 avian influenza virus replication in chick embryo. Animal experiment suggests that recombinant protein is of good antivirus activities.

关 键 词：鸡α干扰素 分泌表达 抗病毒 

分 类 号：S852.65[农业科学—基础兽医学]