高邮麻鸭白细胞介素-2基因的克隆与序列分析  

Cloning and Sequencing of IL-2 Gene in Gaoyou Duck

作　　者：陈诚[1] 佘步宏[1] 任春芝[1] 陶建平[1] 许金俊[1] 

机构地区：[1]扬州大学兽医学院,江苏扬州225009

出　　处：《安徽农业科学》2010年第28期15670-15672,共3页Journal of Anhui Agricultural Sciences

基　　金：江苏省自然科学基金(BK2009701);2009年度扬州大学教学改革研究基金项目资助

摘　　要：[目的]对高邮麻鸭白细胞介素-2(CIL-2)基因进行克隆和序列分析。[方法]以高邮麻鸭外周血淋巴细胞提取的总RNA为模板,根据已发表的鸭IL-2基因cDNA序列设计并合成1对引物,应用两步RT-PCR技术扩增出IL-2基因的特异性片段。将扩增片段插入pGEM-T-easy载体,并进行测序分析和结果验证。[结果]阳性克隆所含插入片段的DNA序列全长为433bp,与预期大小一致,含有1个423bp大小的开放性阅读框,共编码141个氨基酸的前体蛋白,N端21个氨基酸形成信号肽,成熟蛋白为120个氨基酸,分子量约为13.66kD,含1个糖基化位点。该序列与绿头鸭、绍兴鸭、固始鸭相应序列的同源性均为99.8%,与广州白鸭的同源性为99.3%,存在少量核苷酸差异。[结论]高邮麻鸭IL-2编码区基因相对高度保守,为其用于临床疾病治疗提供了一定参考。[Objeotive]To clone and analgze the IL-2 gene in Gaogou duk.[Method]The IL-2 gene of gaoyou duck gene was amplificated by reverse transcription-polymerase chain reaction（RT-PCR） from lymphocytes which stimulated by PHA （phytohaemagglutinin,PHA） for 7 h.The gene was cloned into pGEM-T-easy vector and identificated by enzyme restriction analysis.Positive recombinant clone was sequenced.[Result] The ORF of the cloned gene was 423 bp,encoded a protein of 141 amino acids.N-part carried 21 amino acids which formed signal peptide.Mature polypeptide has 120 amino acids,the molecular weight was approximately 13.66 kD,including one glycosylation position spot.The homology was 99.8% compared with Anas platyrhynchos duck,Gushi duck and Shaoxing duck,but was 99.3% with the Guangzhou white duck.[Conclution]The coding region of IL-2 gene of gaoyou duck was highly conservative.

关 键 词：高邮麻鸭 白细胞介素-2基因 克隆 序列分析 

分 类 号：S852.4[农业科学—基础兽医学]