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| 天府肉鹅IFN-α成熟肽基因的原核表达及抗病毒活性 | |
| 引用本文： | 刘菲,高丽芹,程安春,汪铭书,韩清林,赵婷婷,唐卫杰,严晓玲.天府肉鹅IFN-α成熟肽基因的原核表达及抗病毒活性[J].四川农业大学学报,2011,29(2):269-273. |
| 作者姓名： | 刘菲 高丽芹 程安春 汪铭书 韩清林 赵婷婷 唐卫杰 严晓玲 |
| 作者单位： | 1. 四川农业大学动物医学院,四川雅安,625014；动物疫病与人类健康四川省重点实验室,四川雅安,625014 2. 四川农业大学动物医学院,四川雅安,625014 3. 四川农业大学动物医学院,四川雅安,625014；四川省大英县畜牧局,四川大英,629118 |
| 基金项目： | 四川省教育厅自然科学重点项目(2006A012)；教育部“长江学者和创新团队发展计划”创新团队项目(IRT0848)；现代农业产业技术体系建设专项资金 |
| 摘 要： | 参照已克隆的天府肉鹅IFN-α完整ORF基因序列设计引物,克隆成熟肽基因,与载体pET32a连接,构建原核表达载体,并在表达菌Rosetta中表达,SDS-PAGE分析鉴定,通过Ni-IMAC亲和层析纯化和稀释透析复性后,采用微量细胞病变抑制法分析测定IFN的抗病毒活性.结果显示,IFN-α成熟肽基因全长486 bp,... |
| 关 键 词： | 天府肉鹅 α干扰素 原核表达 抗病毒活性 水泡性口炎病毒 |
Prokaryotic Expression of Tianfu Goose IFN-α Mature Peptide and its Activity Against Virus |
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| LIU Fei,GAO Li-qin,CHENG An-chun,WANG Ming-shu,HAN Qing-lin,ZHAO Ting-ting,TANG Wei-jie,YAN Xiao-ling.Prokaryotic Expression of Tianfu Goose IFN-α Mature Peptide and its Activity Against Virus[J].Journal of Sichuan Agricultural University,2011,29(2):269-273. | |
| Authors: | LIU Fei GAO Li-qin CHENG An-chun WANG Ming-shu HAN Qing-lin ZHAO Ting-ting TANG Wei-jie YAN Xiao-ling |
| Abstract: | A primer was designed reference to the complete ORF of Tianfu goose IFN-α,the mature peptide was cloned and connected into pET32a to construct the prokaryotic expression vector.Then the IFN-α gene was expressed in Rosetta and identified by SDS-PAGE.After the expressed protein was purified by Ni-IMAC affinity chromatography and dialyzed to renaturation,its antiviral activity was determinated by micro cytopathic effect inhibition.The results revealed that IFN-α mature peptide gene was 486 bp and encoded a polypeptide of 161 amino acid.The prokaryotic expression vector pET32a-mGoIFN-α was constructed successfully and the gene was expressed.SDS-PAGE and Western-blotting assays demonstrated the expressed protein was about 38 ku.After purifying and renaturating,the antiviral activity of IFN was evaluated by using vesicular stomatitis virus(VSV) and had an antiviral specific activity of 5.52×103 U/mg,which Showed that the recombinant protein had a good antivital activity. |
| Keywords: | Tianfu goose IFN-α prokaryotic expression antiviral activity VSV |
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