坝上长尾鸡IL-2基因克隆与生物信息学分析

Bioinformatics Analysis of Bashang Long Tail Chicken IL-2 Gene

坝上长尾鸡是河北省唯一被列入家禽志的地方种鸡,为更好地保护此地方品种,应用反转录多聚酶链反应(RT-PCR)技术,对坝上长尾鸡的IL-2基因进行研究。根据GenBank中发表的鸡白细胞介素2(IL-2)基因序列的开放阅读框(ORF)设计1对引物,从坝上长尾鸡脾脏中扩增其IL-2基因,连接pMD18-T载体,并进行序列的测定和分析。结果显示:该基因包含一个开放阅读框为432 bp,编码143个氨基酸。该基因序列与GenBank中发表的鸡、Leghorn SPF鸡、Kestrel来航鸡、broiler鸡、Obese鸡、萧山鸡、猫、牛、山羊、人的IL-2核苷酸序列同源性分别为95.0%,98.1%,95.0%,94.1%,93.9%,94.1%,45.3%,48.0%,47.5%,41.7%;与其氨基酸序列同源性分别为97.2%,97.2%,96.7%,8.1%,9.6%,94.7%,12.3%,14.1%,17.1%,10.2%。这表明坝上长尾鸡IL-2基因被成功克隆,在种属间存在差异性。

Long tail chicken is the only poultry to be included in the local breeding breeders in Hebei Prov- ince. In order to better protect the local breeds, the application of reverse transcription polymerase chain reac- tion （RT-PCR） was used to research IL-2 gene. Based on the published ORF of chicken IL-2 gene, a pair of primers were designed. The IL-2 gene was amplified from the spleen of the Bashang long tail chicken and con- nected to the pMD18-T vector, then its sequence was determined and analyzed. The results showed that the gene included an open reading frame of 432 bp, encoding 143 amino acids. The nucleotide sequence of IL-2 homologous sequences and GenBank gene published in the Leghorn SPF Kestrel chicken, Kestrel chicken, broiler chicken, Obese chicken, Xiaoshan chicken, cat, cow, goat and human was respectively 95.0%, 98.1%, 95.0%, 94.1%, 93.9%, 94.1%, 45.3%, 48.0%, 47.5%, 41.7%. The amino acid sequence homology was respectively 97.2%, 97.2%, 96.7%, 8.1%, 9.6%, 94.7%, 12.3%, 14, 1%, 17.1%, 10.2%. This indicated that the IL-2 gene was successfully cloned, and there were differences among species. This study laid the foundation for further research on the biological function of IL-2 gene, especially the use of IL-2 gene to enhance the immune effect of DNA vaccine.