固始鸡白细胞介素18全基因的克隆与序列分析

Cloning and sequence analysis of the Gushi chicken interleukin-18

根据GenBank发表的鸡IL-18cDNA基因序列,自行设计1对引物,应用RT-PCR技术,无需用非特异性免疫原如ConA,PHA等刺激分离的脾淋巴细胞,直接提取总RNA,扩增固始鸡IL-18基因,并进行克隆和测序.测序结果:获得了固始鸡IL-18基因全序列,其大小为597 bp;序列比较分析发现,克隆的固始鸡IL-18基因与Schneider报道的鸡IL-18基因序列核苷酸同源性为99.8%,有1个碱基发生非同义变异,氨基酸同源性为99.5%;固始鸡与人和其他动物的IL-18基因序列分析和系统进化分析表明,IL-18基因存在种属的差异性,亲缘关系越近,同源性越高.

One pair of primers were designed and synthesized according to chicken IL-18 gene sequences published in GenBank. Gushi chicken IL-18 gene was amplified by RT-PCR from chicken splenocyte total RNA extract, then was cloned and sequenced. The result showed that the full length sequence of Gushi chicken IL-18 gene consisted of 597 bp. When the nucleotide sequence of Gushi chicken IL-18 gene compared with that of chicken IL-18 genes published in GenBank, the nucleotide sequence shared 99.8% homology with the nucleotide sequence published by Schneider. The results of comparative sequence analysis and phylogenetic tree analysis indicated there was difference of genus in IL-18 gene, the nearer relationship, the higher homology.