猪白细胞介素-10的分子克隆与序列测定  

Cloning and Sequencing of Porcine IL-10

作　　者：兰文升[1] 童光志[1] 仇华吉[1] 王云峰[1] 周艳君[1] 郭海龙[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2002年第2期113-115,共3页Chinese Journal of Preventive Veterinary Medicine

摘　　要：从LPS活化的猪外周血单个核细胞 (PBMC)提取总RNA ,用RT_PCR方法扩增出猪白细胞介素_10 (IL_10 )的编码基因 ,将其连接到pUC18质粒上 ,然后进行BamHI和HindIII双酶切鉴定筛选阳性克隆 ,序列分析结果表明该基因开放阅读框由5 2 8个核苷酸组成 ,推测产生的编码产物由 175个氨基酸组成。猪IL_10基因与人、大鼠和小鼠基因序列的同源性分别为84%、79%和 79% ,与国外报道的猪IL_10基因序列完全一致 。The total RNA was extracted from pig peripheral blood mononuclear cells (PBMCs) stimulated by LPS.Porcine IL-10 gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR).The gene was cloned into pUC18 vector and identified by PCR and digestion by BamHI and HindIII.Then one of the positive recombinant clones was sequenced and analyzed.The result showed that the gene has only one ORF,which is composed of 528 nucleosides.The predicted peptide included 175 amino acids.It also showed that the sequence was 84%,79% and 79% identity to that of human,rat and mouse,respectively.The sequence was just the same as the published one from GenBank.So the porcine IL-10 was cloned from porcine PBMCs successfully.

关 键 词：猪 白细胞 介素-10 分子克隆 序列测定 

分 类 号：S828[农业科学—畜牧学] Q813[农业科学—畜牧兽医]