广西沼泽型水牛IL-4和IL-5基因的克隆及其序列分析

Cloning and sequencing of Guangxi swamp type buffalo IL-4 and IL-5

从刀豆素A(Concanavalin A,ConA)刺激的广西沼泽型成年水牛外周血单个核细胞(Peripheralblood mononuclear cells,PBMCs)中提取总RNA,用逆转录聚合酶链式反应(RT-PCR)方法扩增出白细胞介素-4(Interleukin-4,IL-4)和白细胞介素-5(IL-5)基因,分别克隆到pMD18-T载体上,进行序列分析。结果表明,从培养9 h、17 h的单个核细胞中扩增得到的IL-4基因在核苷酸水平上与GenBank上登录的印度水牛IL-4 cDNA序列同源性为98.8%,与牛、绵羊、猪和马的同源性分别为99.3%、94.1%、84.8%和80.6%;氨基酸水平的同源性分别为96.3%、98.5%、90.4%、78.4%和59.6%。从培养12 h的单个核细胞中扩增得到的IL-5基因在核苷酸水平上与GenBank上登录的牛、绵羊、猪和马IL-5 cDNA序列同源性分别是99.0%、96.5%、89.6%和89.1%;氨基酸水平的同源性分别为97.8%、96.2%、85.9%和86.7%。本研究为进一步了解水牛IL-4和IL-5的结构和水牛免疫学特性奠定了基础。

The cDNAs encoding interleukin-4 （IL-4） and interleukin-5 （IL-5） of Guangxi swamp type buffalo were amplified by reverse transcription polymerase chain reaction （RT-PCR） from concanavalin A （ConA） stimulated buffalo peripheral blood mononuclear cells （PBMCs）. The RT-PCR products were cloned into pMDI 8-T vector, and sequenced. IL-4 gene cloned from PBMCs incubated for 9 h and 17 h shares 98.8 %, 99.3 %, 94.1%, 84.8 %, 80.6 % nucleotide homology and 96.3 %, 98.5 %, 90.4 %, 78.4 % and 59.6 % amino acid homology with those of buffalo （India）, bovine, sheep, pig and horse, respectively. IL-5 gene cloned from PBMCs incubated for 12 h shared 99.0 %, 96.5 %, 89.6 % and 89.1% nucleotide homology, 97.8 %, 96.2 %, 85.9 % and 86.7 % amino acid homology with that of bovine, sheep, pig and horse, respectively. This study provided foundation for further study on structure of buffalo IL-4 and IL-5 in buffalo.