中国农业科学院农业信息研究所 中国农学会
Agricultural Information Institute of CAAS China Association of Agricultural Science Societies
En
Ch
中国农业科学院农业信息研究所 中国农学会 主办

AI检索

订阅

返回顶部
当前位置: 首页 > 文章 > 鸡γ-干扰素在毕赤酵母中的分泌表达及其抗病毒活性研究 中国家禽 2017,39 (1) 30-33
Position: Home > Articles > Expression of Chicken Interferon-γ in Pichia pastoris and Its Antiviral Activity China Poultry 2017,39 (1) 30-33

鸡γ-干扰素在毕赤酵母中的分泌表达及其抗病毒活性研究

作  者：
刘新文;王秀丽;郭东春;邹敏;郭伟伟;宫晓;李陆梅;范根成
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γ-干扰素;分泌表达;抗病毒活性
摘  要：
将克隆的鸡γ-干扰素(Ch IFNγ)基因亚克隆至酵母表达载体p PICZα-A中,构建重组质粒p PIC-Ch IFNγ。将鉴定正确的质粒p PIC-Ch IFNγ线性化,通过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,表达产物经SDS-PAGE检测,结果表明Ch IFNγ在酵母中获得了分泌型表达,表达产物分子量约为20 ku。表达产物粗提后以细胞病变抑制法检测其活性,效价为107.69U/m L;在鸡胚中能有效抑制H9N2亚型禽流感病毒的增殖;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-γ in Pichia pastoris and Its Antiviral Activity
作  者：
LIU Xinwen;WANG Xiuli;GUO Dongchun;ZOU Min;GUO Weiwei;GONG Xiao;LI Lumei;FAN Gencheng;The Qingdao Yebio Biological Engineering Co.,Ltd.;Harbin Veterinary Research Institute,CAAS;
关键词：
chicken interferon-γ;;secreted expression;;antiviral activity
摘  要：
The gene of chicken interferon gamma(Ch IFNγ)was amplified by reverse transcription- polymerase chain reaction(RT-PCR)from SPF chicken blood genome and cloned. The coding region(495 bp)was subcloned into the expression vector p PICZα-A. The recombinant plasmid p PIC-Ch IFNγ was identified by enzyme digestion. After linearized by SacⅠ,the plasmid was transferred into Pichia pastoris strain X-33.The recombinant strain was isolated and identified by PCR. The recombinant strain was induced by 1% methanol for 4 days,the production of recombinant Ch IFNγ was detected by SDS-PAGE. The assay results showed that the protein weight was approximate 20 ku,and the recombinant protein was secreted effectively in Pichia pastoris. Antiviral activity of the recombinant Ch IFNγ was detected by the classical experiment of cell pathological effect inhibition assay. The results showed the titer was about107.69U/m L;The expressed protein could also inhibit subtype H9N2 avian influenza virus replication in chick embryo and SPF chick;Animal experiment indicated that recombinant protein had good antivirus activity.

相似文章

导出引用
计量
文章访问数: 8
HTML全文浏览量: 0
PDF下载量: 0

所属期刊

推荐期刊

友情链接Friendly link
农业信息研究所Agricultural information institute 农业智能知识服务平台Agricultural intelligent knowledge service platform 农业学术服务平台Agricultural Knowledge Service System 中国知网CNKI 维普网VIP 万方WANFANG DATA
联系我们Contact us

电话Tell：010-82109652-808

邮箱E-mail：agrijournal@caas.cn

服务Service
关于我们About us 意见建议Opinions and suggest
版权说明Copyright
单位：中国农业科学院农业信息研究所 中国农学会 © 2017中国农业科学院农业信息研究所 ICP备案号：京ICP备09089781号-46