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| 猪IFN-α在巴斯德毕赤酵母中的分泌表达 | |
| 引用本文： | 林宝珍,朱艳平,黄文科,岑路,郭霄峰.猪IFN-α在巴斯德毕赤酵母中的分泌表达[J].华南农业大学学报,2010,31(1). |
| 作者姓名： | 林宝珍 朱艳平 黄文科 岑路 郭霄峰 |
| 作者单位： | 1. 南海出入境检验检疫局,广东,南海,528200 2. 华南农业大学,兽医学院,广东,广州,510642 3. 广州粤丰动物保健有限公司,广东,广州,510830 |
| 基金项目： | 广东省动物防疫检疫研究专项经费项目 |
| 摘 要： | 为了获得高效抗病毒活性的猪α-干扰素蛋白,采用PCR法扩增获得猪α-干扰素基因完整的开放阅读框(ORF),构建重组表达质粒pPICZαC-IFN,电击转化毕赤酵母感受态细胞X33.挑阳性菌落诱导表达,离心取上清,SDS-PAGE和Western-blotting分析,可见1条相对分子质量约19 400的清晰蛋白条带,与理论值相符,表明表达产物为猪α-干扰素.在Vero细胞上检测该干扰素抗VSV的活性为4.04×106IU/L. |
| 关 键 词： | 猪a-干扰素 巴斯德毕赤酵母 分泌表达 抗病毒活性 |
The Secretive Expression of Porcine Alpha-Interferon Gene in Pichia pastoris |
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| LIN Bao-zhen,ZHU Yan-ping,HUANG Wen-ke,CEN Lu,GUO Xiao-feng.The Secretive Expression of Porcine Alpha-Interferon Gene in Pichia pastoris[J].Journal of South China Agricultural University,2010,31(1). | |
| Authors: | LIN Bao-zhen ZHU Yan-ping HUANG Wen-ke CEN Lu GUO Xiao-feng |
| Abstract: | In order to get alpha-interferon(INF-α) with high level secretive expression and high antiviral activities,the porcine alpha-interferon gene was amplified by PCR,and a DNA fragment of 501 bp,the open reading framework (ORF) was obtained.Then,the target gene and pPICZαC were digested with EcoRI/XbaI and were linked.The recombinant plasmid of pPICZαC-IFN was linearized by SacI and electroporated into Pichia pastoris X-33.PCR assay was used to identify colonies.The high copy recombinat strains were screened and induced by regulation of methanol utilization.IFN-α protein was detected by SDS-PAGE and Western-blotting analysis.The result showed the IFN-α protein with a raletive molecular mass of 19 400 was expressed in Pichia pastoris X-33.The antiviral activity of IFN-α against vesicular stomatitis virus(VSV) was investigated on the Vero cell and the result indicated that IFN-α could inhibit VSV and the activity was 4.04×10~6 IU/L. |
| Keywords: | porcine alpha-interferon Pichia pastoris secrete expression anti-virus activity |
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