三元杂交猪IL-18全基因的序列测定及分析

摘要

[Objective] To clone the porcine interleukin-18(IL-18) cDNA and explore the immunological effectiveness of porcine IL-18 as an adjuvant of genetic vaccine. [Method] The spleen lymphocytes were isolated from Henan three-way cross-breeding pigs. According to the porcine IL-18 gene in GenBank, a pair of specific primers was designed. The full length cDNA of porcine IL-18 was amplified by RT-PCR. Subsequently, porcine IL-18 cDNA was cloned into pGEM-T vector and sequenced and analyzed. [Result] The porcine IL-18 gene demonstrated an open reading frame of 579 bp encoding an inactive precursor protein with 192 amino acids. The precursor protein had no typical hydrophobic signal peptide and cleaved by interleukin-1 beta (IL-1β) converting enzyme (ICE) in caspase-1 splice site; the porcine mature protein had biological activity. After comparing with other porcine IL-18 genes, the nucleotide sequence homology was over 96% and the deduced amino acid homology was more than 98%. [Conclusion] A full length procine IL-18 gene was gained. It lays the foundation for porcine IL-18 as an adjuvant of genetic vaccine.