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%。

关键词: 猪, 白细胞介素-2, 基因, 克隆, 遗传进化分析

Abstract: 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 μ g/mL ConA in vitro for 27 h. Then the amplified cDNA was cloned 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%, respectively.

Key words: Procine, Interleukin -2, Gene, Cloning, Phylogenetic analysis

中图分类号: 

• Q785