貉白介素－18(IL-18)基因的克隆及序列分析

To provide a basis for further investigation of the function of Raccoon dog IL-18, a pair of primers specific to Raccoon dog Interleukin-18(IL-18) were designed and synthesized according to the known dog IL-18 sequence from GenBank. Raccoon dog IL-18 cDNA were used to amplify by RT-PCR from total RNA of Raccoon dog PBMC cells induced with PHA.RT-PCR product was cloned into pMD18-T vector. The nucleotide acid sequence was determined after being identified by restriction endonuclease digestion. The result showed that cloned Raccoon dog IL-18 cDNA was 590 bp in length with a 582 bp open reading frame encode a polypeptide of 193 amino acid. Compared with published arctic fox and dog IL-18, nucleotide acid homology were 95.9% and 94.8%, and amino acid homology were 93.8% and 91.2%, respectively. There is significant difference between Raccoon dog and other species of IL-18 gene sequence. The obtaining of IL-18 sequence laid the foundation for its further study and exploitation.