犬白细胞介素-21基因的扩增和测序  

Amplification and Sequencing of Canine IL-21 cDNA

作　　者：张晓东[1] 栾维民[1] 钱爱东[1] 

机构地区：[1]吉林农业大学动物科技学院,长春130118

出　　处：《吉林农业大学学报》2006年第4期450-452,共3页Journal of Jilin Agricultural University

基　　金：国家自然科学基金资助项目(30070542)

摘　　要：无菌抽取犬外周血,分离单核细胞,经PHA刺激后收集细胞,使用Trizol提取细胞总RNA;利用自行设计的引物,反转录合成cDNA第1条链,PCR扩增出长约441 bp的白细胞介素-21(IL-21)基因带。经纯化回收,将基因克隆至pMD18-T,筛选阳性克隆测序证明扩增的cDNA的序列与已报道的序列相一致。In this study, canine interleukin-21 cDNA was amplified. Canine leukocytes were separated from ,peripheral blood stimulated with PHA for 12 - 24 h. Total RNA was extracted from PHA-aetivated canine T cells by using Trizol. The full length eDNA of canine IL-21 gene was amplified by RT-PCR using leukocyte mRNA as a template and designed primer, which was proven to be about 441bp through agarose gel eleetrophoresis. Then IL-21 eDNA was cloned into pMD18-T. Sequencing result showed that the full length eDNA was 441bp, which was identical with that published. Target gene was successfully amplified by RT-PCR and was proven by sequencing.

关 键 词：白细胞介素-21(IL-21) 基因 RT-PCR T细胞 犬 

分 类 号：S858.292[农业科学—临床兽医学]