罗曼鸡胚脾细胞白介素-18基因的克隆与序列分析

Cloning and Sequencing of IL-18 cDNA from Chicken Embryo Spleen

根据国外已发表的鸡白细胞介素 18(IL- 18) c DNA基因序列设计了 1对特异性引物 ,应用 RT- PCR技术 ,从鸡新城疫 系病毒接种 4 8h左右的罗曼鸡胚脾细胞中扩增出鸡 IL - 18全基因 ,并进行了序列测定。结果表明 ,扩增片段全长 5 94 bp,共编码 198个氨基酸的前体蛋白 ,其中含有表达完整功能蛋白所必需的起始密码子和终止密码子。该序列与国外报道的鸡 IL - 18全基因核苷酸序列及推导的氨基酸序列的同源性分别为 99.8%和 10 0 % ;序列中编码成熟蛋白的这段基因与国内报道的源自白来航鸡编码 IL- 18成熟蛋白的基因核苷酸序列及推导的氨基酸序列的同源性分别为 99.8%和 99.4 %。本研究为鸡 IL - 18的扩增及其他细胞因子的扩增提供了一种简便易行的新方法 ,为进一步研究IL- 18基因的结构、功能。

A pair of primers specific to chicken IL-18 were designed and synthesized according to the previously published sequence,to amplify chicken IL-18 cDNA by reverse transcription polymerase chain reaction(RT-PCR) from spleen cells of chicken embryo inoculated with Newcastle disease virus.Sequence analysis showed that the IL-18 cDNA was 594 bp in length,and encoded a protein of 198 amino acids.Compared with the chicken IL-18 cDNA reported by Schneider K,the homology of nucleotide acid and amino acid were 99.8% and 100%,respectively;the mature peptide gene homology of nucleotide and amino acid were 99.8% and 99.4%.This study is the first report of chicken IL-18 complete gene in China,it will lay a sound foundation for further study on the structure,function,expression of chicken IL-18.In addition,it also provide a new way for amplify chicken IL-18 and other cytokine.