猪α干扰素基因在pET32a表达载体中的表达和鉴定

IFN-α of Pig gene expression and identification in pET32a expression vector

利用PCR技术从猪瘟免疫猪血中提取基因组,并从中扩增出约500bp的基因片段;将此基因片段克隆于大肠杆菌pMD18-T载体,经PCR、测序鉴定后,将其片段克隆于pET32a(+)载体进行诱导表达,表达产物经SDS-PAGE检测,结果表明,猪α干扰素在pET32a原核表达载体中成功地获得了分泌表达,且表达蛋白在PK-15细胞上表现出较强的抗病毒活性,这为进一步研究猪α干扰素的功能和开发奠定了基础。

To extract genome the blood of pigs immunized with classical swine fever vaccine, which was a gene fragment of about 500bp was amlified .by PCR; this gene fragment was cloned into E. coli pMD18-T vector, after PCR, DNA sequencing, the fragment was cloned PET32a（＋） vector, expression products were detected by SDS-PAGE. The results show that the swine interferon-α was expressed using pET32a prokaryotic expression vector, and the expression protein in PK-15 cells showed strong antiviral activity, which laid the foundation for further study of the development and function of the IFN-α of Pig.