长白猪γ-干扰素基因的克隆与序列分析  

Cloning and Sequence Analysis of Changbai Porcine Interferon-γ Gene

作　　者：赵英杰[1] 徐宏军[1] 李尚波[1] 王文成[1] 宣华[2] 

机构地区：[1]辽宁省益康生物制品厂研发中心,辽宁辽阳111000 [2]吉林大学农学部军事兽医系,吉林长春130062

出　　处：《动物医学进展》2005年第7期49-53,共5页Progress In Veterinary Medicine

摘　　要：将长白猪外周血淋巴细胞进行体外培养,在ConA的刺激下培养8h～24h后,提取培养的淋巴细胞的总RNA,应用RTPCR技术扩增淋巴细胞γ干扰素cDNA,并将其克隆到pGEMT载体上,进行测序。序列测定结果表明,克隆获得的IFNγ基因全长为534个碱基,ORF为501个碱基,编码166个氨基酸,分子质量为19.4ku。此基因与我国藏猪、成华猪以及GenBank上发表的猪IFNγ基因的核苷酸同源性均为100%,并且在氨基酸水平上也完全一致。说明几种我国地方猪IFNγ在基因的核苷酸以及蛋白的氨基酸水平上不存在差异和突变,同时也证实了长白猪干扰素γ基因的正确、完整克隆。Peripheral blood lymphocytes isolated from Changbai porcine were cultivated in RPMI-1640 culture medium, Then Concavadin A was used to stimulate the peripheral blood lymphocytes for 8-24 h. After the total RNA isolated from the lymphocyte, the PoIFN-γ gene was amplified via RT-PCR.The gene segment of IFN-γwas cloned into the vector pGEM-T. From the sequence,it was found that the gene of Changbai PoIFN-γ was consisted of 536 nucleotides,The ORF was consisted of 501 nucleotides, encoding 166 amino acids, molecular weight of the protein is 19.4 ku. The homology of Changbai porcine IFN-γ gene was 100% at nucleotide level and amino level among Chenghua porcine、Zang porcine and the published sequence on Genebank. The results showed that Changbai PoIFN-γ gene and the induced protein have no difference with some local breeds of porcine. And proved that the gene was cloned correctly and completely.

关 键 词：长白猪 Γ-干扰素基因 克隆 序列分析 

分 类 号：Q785[生物学—分子生物学]