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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志  2006, Vol. 26 Issue (0): 113-117    
专稿     
口蹄疫病毒3C蛋白酶基因克隆及与SOC的融合表达
田纯见 毕英佐 曹永长 马静云 谢青梅
华南农业大学动物科学学院 华南农业大学动物科学学院 华南农业大学动物科学学院 华南农业大学动物科学学院 华南农业大学动物科学学院
Cloning of the Foot and Mouth Disease Virus 3 C Gene and Expression of Its SOC Fusion Product
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摘要:

本文通过RT-PCR技术,从乳鼠胴体材料中扩增出完整的3C基因片段,克隆到pGEM T easy载体中,经测序鉴定获得含3C基因的重组质粒p T-3C。用EcoR I单酶切技术构建3C蛋白酶融合表达载体,经引物对Pr78/3C-A2筛选3C片断插入方向正确的克隆子。所构建的原核表达载体pSOC-3C经酶切鉴定后,在IPTG诱导下,大肠杆菌E.coli BL21 (DE3)plys S在26~37℃范围内可表达SOC-3C融合蛋白,出现约26KD大小的目的条带,经Western-blot 鉴定,具有免疫反应性。研究结果为进一步开展3C蛋白酶的噬菌体表面展示和新型疫苗、药物研究打下了基础。

Abstract:

Using RT-PCR technique, the entire FMDV-3C gene segment was amplified from suckling mouse carcass and cloned into pGEM T easy vehicle to obtain the recombinant plasmid pT-3C, which was confirmed by nucleotide sequencing. EcoRI cleavage was used to construct a 3C protease fusion expression vehicle, and recombinants having the correct orientation were screened with a pair of primers Pr78/3C-A2. The prokaryotic expression vehicle pSOC-3 C constructed was subjected to restriction analysis and, under the induction of IPTG, could in E.coli BL21 (DES)plysS express the SOC-3C fusion protein that gave a 26 KD-sized target electrophoretic band, the latter exhibiting immunoreactivity on Western-blotting. These findings paved the way for further research in bacteriophage surface display of the 3C protease and development of novel vaccines and pharmaceuticals

收稿日期: 2006-01-23 出版日期: 2006-06-15
通讯作者: 毕英佐   
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引用本文:

田纯见,毕英佐,曹永长,马静云,谢青梅. 口蹄疫病毒3C蛋白酶基因克隆及与SOC的融合表达[J]. 中国生物工程杂志, 2006, 26(0): 113-117.

. Cloning of the Foot and Mouth Disease Virus 3 C Gene and Expression of Its SOC Fusion Product. China Biotechnology, 2006, 26(0): 113-117.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/        https://manu60.magtech.com.cn/biotech/CN/Y2006/V26/I0/113

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