Abstract:

Abstract：A pair of specific primer for porcine interferon-alpha1 (poIFN-α1) was designed and synthesized according to gene sequence (EU364896), and poIFN-α1 gene was amplified by RT-PCR from peripherals blood lymphocytes (PBLC) of ternary hybrid piglet.Then poIFN-α1 complete gene was cloned into pGEM-T vector and sequenced. Moreover, poIFN-α1 gene with enzyme sites was cloned into prokaryotic expression vector pGEX-6P-1 to construct recombinant expression plasmid pGEX-6P-poIFN-α1 and transform it into E.coli BL21 (DE3). Recombinat poIFNα1 was expressed and identified with expression conditions confirmed. The analysis of sequence indicated that the cloned poIFN-α1gene consisted of 546 bp encoding 181 aa with a signal peptide of 23 aa but without glycosylation site. The selected recombinant expressed approximately a 46 ku fusion protein with glutathione S-transferase (GST) and poIFN-α1 which was demonstrated by SDS-PAGE and Western blotting. The amount of fusion protein reached the highest when the recombinants were induced for 9 hours on 37℃ with 1.0 mmol/L IPTG. Clone and expression of poIFN-α1 gene laid a foundation for the study of antivirual activity and utilization of poIFN-α1.