奶牛白细胞介素2基因的克隆及序列分析  

Cloning,Identification and Sequence Analysis of the Bovine Interleukin-2 Gene

作　　者：盖仁华[1] 平丽[1] 李广兴[2] 

机构地区：[1]浙江大学药物安全评价研究中心,浙江杭州310058 [2]东北农业大学动物医学学院,黑龙江哈尔滨150030

出　　处：《动物医学进展》2012年第11期17-21,共5页Progress In Veterinary Medicine

基　　金：黑龙江省自然科学基金重点项目(ZJN0702-01)

摘　　要：根据GenBank发表的奶牛白细胞介素2(IL-2)基因序列,设计一对特异性引物,应用RT-PCR技术扩增奶牛IL-2基因,琼脂糖电泳显示扩增出的片段约为477bp。回收片段,克隆入pMD18-T载体,经菌落PCR、酶切及重组质粒PCR鉴定,测序结果显示,克隆的奶牛IL-2基因与GenBank发表的奶牛IL-2基因序列同源性为98.7%。该序列与水牛和山羊的IL-2基因序列同源性最大,在95%以上;氨基酸和抗原性比较分析显示,奶牛的IL-2与山羊及水牛的IL-2在这两方面有较高的同源性。Based on the published nucleotide sequence of bovine interleukin-2,a pair of RT-PCR primers were designed and synthesized.Total bovine cell RNA,isolated from ConA-stimulated peripheral blood of bovine,was used as template to generate complementary DNA by reverse transcription.The 0.477 kb DNA fragment was amplified by polymerase chain reaction（PCR）,and cloned into the pMD18-T vector.DNA sequencing confirmed the fragment was bovine interleukin-2 and the sequence was identical to the sequence of IL-2 publised in GenBank;Sequence analysis showed that the cloned sequence has very high similarity with that of ovine and Bubalus bubalis IL-2,over 95%;Comparative analysis of both amino acid sequence and antigenecity showed that the interleukin-2 of bovine has high similarity to the one of ovine and Bubalus bubalis.

关 键 词：白细胞介素2基因 克隆 序列分析 奶牛 

分 类 号：S852.4[农业科学—基础兽医学] S858.23[农业科学—兽医学]