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当前位置: 首页 > 文章 > 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 中国兽医科学 2017 (5) 626-629
Position: Home > Articles > Expression of chicken interferon alpha in Pichia pastoris and studies on antiviral effects Chinese Veterinary Science 2017 (5) 626-629

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

作  者：
王秀丽;徐慧琳;刘新文;郭东春;郭伟伟;邹敏;李陆梅;杜元钊
单  位：
中国农业科学院哈尔滨兽医研究所;青岛易邦生物工程有限公司
关键词：
鸡α干扰素;分泌表达;抗病毒
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。
译  名：
Expression of chicken interferon alpha in Pichia pastoris and studies on antiviral effects
作  者：
WANG Xiu-li;XU Hui-lin;LIU Xin-wen;GUO Dong-chun;GUO Wei-wei;ZOU Min;LI Lu-mei;DU Yuan-zhao;The Qingdao Yebio Biological Engineering Co.,Ltd.;Harbin Veterinary Research Institute,Chinese Academy of Agricultural Science;
关键词：
chicken interferon-α;;secreted expression;;antiviral activity
摘  要：
The gene of chicken interferon alpha(ChIFN-α) was was subcloned into the expression vector pPICZα-A.After linearized by SacⅠ,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-α 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-α 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.

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