四川农业大学学报 ›› 2006, Vol. 24 ›› Issue (02): 210-213.doi: 10.16036/j.issn.1000-2650.2006.02.019

• 研究论文 • 上一篇    下一篇

猪传染性胃肠炎病毒SC-Y株N基因的克隆及原核表达

宋振辉, 郭万柱, 韩国全, 骆爱芳   

  1. 四川农业大学 动物生物技术中心, 四川 雅安 625014
  • 收稿日期:2005-09-22 出版日期:2006-06-30 发布日期:2017-03-06
  • 基金资助:
    教育部长江学者和创新团队发展计划资助(IRT0555)

Cloning and Prokaryotic Expression of Nucleocapsid Protein Gene of Transmissible Gastroenteritis Virus SC-Y Strain

SONG Zhen-hui, GUO Wan-zhu, HAN Guo-quan, LUO Ai-fang   

  1. Animal Biology Technology Center, Sichuan Agricultural University, Yaan 625014, Sichuan, China
  • Received:2005-09-22 Online:2006-06-30 Published:2017-03-06

摘要: 通过PCR方法扩增猪传染性胃肠炎病毒SC-Y株核衣壳蛋白(N)基因,然后将重组到pMD18-T载体中的约1174 bp的基因片段亚克隆到PET-32a(+)表达载体上,通过酶切及PCR鉴定阳性的重组质粒命名为PET-N,核苷酸及推导的氨基酸序列分析表明,该基因与其他猪传染性胃肠炎病毒相应基因具有很高的同源性,将阳性重组质粒转化BL21(DE3),经IPTG诱导可表达分子量约66 kD的融合蛋白,N基因的表达可为传染性胃肠炎病毒的诊断提供良好的物质材料。

关键词: 猪传染性胃肠炎病毒, SC-Y株, N基因, 克隆, 原核表达

Abstract: The nucleocapsid protein gene of transmissible gastroenteritis virus SC-Y strain was amplified by reverse transcription-polymerase chain reaction(RT-PCR). The recombinated fragment in pMD18-T was subcloned into the prokaryotic expression vector PET-32a(+). The recombinant expression vector that contained the target fragment was identified by using restriction endonucleases and named as PET-N. The nucleotide and deduced amino acid sequences of the cloned N gene from ransmissible gastroenteritis virus SC-Y strain were analyzed. The results show that the gene has a relatively high homology with the corresponding gene from other transmissible gastroenteritis virus strains. The recombinant plasmid is transformed into BL21 and the recombinant bacteria is induced by IPTG. SDS-PAGE are performed to detect the N fusion protein. Result indicates the molecular weight of N fusion protein is 66 kD. The expression of N protein is to provide a basis for the research on diagnosis of TGEV. biological functions. The constucton of recombinant N gene.

Key words: transmissible gastroenteritis virus, SC-Y strain, N gene, cloning, prokaryotic expression

中图分类号: 

  • S852.659.6