淮南猪IL-2全基因的克隆与遗传进化分析  

Cloning and Genetic Analysis on Interleukin-2 Gene of Huainan Pig

作　　者：郭小参[1] 崔保安[1] 陈红英[1] 杨明凡[1] 管倩[1] 吕晓丽[1] 王东方[1] 

机构地区：[1]河南农业大学牧医工程学院

出　　处：《华北农学报》2008年第4期14-18,共5页Acta Agriculturae Boreali-Sinica

基　　金：国家“十五”食品安全重大攻关专项(2001BA804A30-11)

摘　　要：参照GenBank发表的猪IL-2cDNA基因序列,设计1对引物,将河南地方品种淮南猪脾淋巴细胞在伴刀豆球蛋白A(Con A)的刺激下体外培养27 h后,提取激活淋巴细胞总RNA,进行反转录-聚合酶链反应(RT-PCR)扩增,克隆到pGEM-T Easy载体上并测序。测序结果显示,克隆的HuainanPIL-2 cDNA全长为516 bp,开放阅读框(ORF)包含465bp,编码154个氨基酸,分子量为17.4 kDa,等电点为5.27,疏水氨基酸38.3%,亲水氨基酸42.0%,碱性氨基酸11%,酸性氨基酸23%。此cDNA与已报道的猪IL-2同源性为100%,与猫、牛、鸡、狗、鸭、山羊、马、人、家鼠等的IL-2基因进行比较分析,核苷酸同源性分别为83.7%,82.6%,28.2%,80.6%,29.6%,83.4%,81.3%,82.0%,61.5%。One pair of primers was designed based on the pig IL-2 gene sequences published in GenBank and used to amplify the interleukin-2 cDNA of Huainan Pig by RT-PCR from total RNA extracted from spleen lymphocytes, which were cultivated and stimulated with 10 μg/mL ConA in vitro for 27 h. Then the amplified cDNA was eloned into pGEM- T Easy vector and sequenced. The result showed that the full length of Huainan pig IL-2 was 516 bp with an ORF of 465 bp,encoding a peptide of 154 amino acids which molecular weight is 17.4 kDa, pI 5.27, hydrophobic amino acid 38.3 %, hydrophilic amino acid 42.0 %, basic amino acid 11% and acidic amino acid 23 %. The sequence was the same as the corresponding genes published in Genebank. The base similarity of pig IL-2 with those of cat, cattle, chicken, dog, duck,goat,horse,human and house mouse was 83.7% ,82.6% ,28.2% ,80.6% ,29.6% ,83.4% ,81.3% ,82.0% and 61.5 %, respeetively.

关 键 词：猪 白细胞介素-2 基因 克隆 遗传进化分析 

分 类 号：Q785[生物学—分子生物学]