稳定表达猪干扰素调节因子7的ST细胞株的建立

Establishment of ST cell line stably expressing porcine interferon regulatory factor 7

干扰素调节因子(IRF)在抗病毒感染中具有重要作用,为构建稳定表达猪IRF7(pIRF7)的猪睾丸细胞系(ST),本研究从猪淋巴细胞中扩增获得pIRF7基因,将其插入pEGFP-C3载体中构建重组表达质粒pEGFP-pIRF7。并转染于ST细胞中,通过G418加压筛选及细胞纯化获得G418抗性ST细胞株(STA1)。RT-PCR和western blot检测结果表明,STA1细胞株能够稳定表达重组pIRF7;而且以聚肌胞刺激STA1细胞可以观察到表达的pIRF7在细胞内具有核转位能力,该项研究为进一步研究猪瘟病毒对pIRF7的作用奠定基础。

Interferon regulatory factors play a vital role in anti-virus process as far as we know. To establish a ST cell line stably expressing porcine interferon regulatory factor 7 （pIRF7）, we cloned pIRF7 gene from porcine lymphocytes and inserted into pEGFP-C3 vector to construct a recombinant plasmid pEGFP-pIRF7. The ST cell line stably expressing pIRF7 was selected from the pEGFP-pIRF7 tansfected ST cell by G418, designated STA1. In addition, the expressions of pIRF7 mRNA and protein in STA1 cell line were detected by RT-PCR and western blot. Moreover, the location of recombinant pIRF7 protein was observed in the STA1 cell stimulated by polyinosinic-polycytidylic acid, which indicated that recombinant pIRF7 was able to translocate into nuclear. These results lay a foundation for study of the interaction of pIRF7 with virus such as classical swine fever virus.