贵州白香猪的白细胞介素2基因的克隆及其序列分析

Cloning and Sequence Analysis of Guizhou Baixiang Pig Interleukin-2(IL-2) Gene

为获得贵州白香猪白细胞介素2基因,以提取的经刀豆蛋白A诱导培养的贵州白香猪外周血淋巴细胞总RNA为模板,扩增全长猪白细胞介素2(IL-2)基因,并克隆至pMD18-T Simple载体后测序。结果表明:贵州白香猪IL-2基因ORF长为465bp,可编码154个氨基酸,其中前20个氨基酸为信号肽,后134个氨基酸为成熟肽,具有一个潜在的N-糖基化位点;贵州白香猪除与FJ543109(99.4%)、Neijiang(98.7%)、JN851821(98.7%)、AB194099(98.5%)氨基酸一致性相对较低外,与其他猪源IL-2的一致性均为100%,而与其他种属动物的IL-2基因氨基酸一致性则较低,为11.9%～75%。

With the purpose of getting interleukin-2 gene of Guizhou Baixiang Pig,the total RNA was extracted from peripheral blood mononuclear cells（PBMC）,which was isolated from Guizhou Baixiang Pig and induced with concanavaline A（ConA）.The porcine interleukin-2 gene was amplified by RT-PCR from the total RNA,and cloned to the pMD18-T Simple vector for sequencing.The results of sequence analysis indicated that Guizhou Baixiang Pig IL-2 gene ORF length was 465 bp,encoding 154 amino acids,of which the first 20 amino acid signal peptide and 134 amino acids of the mature peptide,and had a potential N-glycosylation site.The amino acid identity of Guizhou Baixiang Pig was relatively lower among the pigs,such as FJ543109（99.4%）,Neijiang（98.7%）,JN851821（98.7%） and AB194099（98.5%）,and was 100% with the other pigs,but with other species of animals IL-2 gene amino acid identity was low,as 11.9 percent to 75 percent.