四川白鹅白细胞介素-2基因的克隆及生物信息学分析

Cloning and Bioinformatics Analysis of Interleukin-of Sichuan White Goose

[目的]克隆分析四川白鹅白细胞介素-2(IL-2)基因。[方法]参照GenBank中发表的鸭IL-2基因保守序列,设计1对引物,采用RT-PCR技术从ConA刺激的四川白鹅外周血淋巴细胞的总RNA中扩增鹅IL-2基因,并进行测序及生物信息学分析。[结果]四川白鹅IL-2基因全长468 bp,含1个441 bp的开放阅读框(ORF),编码146个氨基酸。生物学软件分析表明该序列编码的氨基酸序列中有4个磷酸化位点,1个糖基化位点,含有21个氨基酸残基组成的信号肽。同源性分析表明四川白鹅IL-2基因与鸭、鸡和火鸡IL-2核苷酸及氨基酸序列同源性均分别为92.2%、77.5%、78.2%和85.8%、65.5%、64.1%,而与其他种属的哺乳动物及啮齿类动物(如人、猴、大鼠、牛、马、猪、猫、小鼠、兔和鹿)的IL-2基因的核苷酸和氨基酸同源性均分别低于45%和28%。[结论]四川白鹅IL-2基因与鸭和鸡具有较近的亲缘关系。

[Objective] The study aimed to clone interleukin-2（IL-2） gene from Sichuan white goose.[Method] Based on the IL-2 gene of duck accessed in GenBank,a pair of primers was designed for cloning IL-2 gene from total RNA of peripheral blood lymphocytes of Sichuan white goose stimulated by ConA via RT-PCR technology.The yielded fragment was sequenced for bioinformatics analysis.[Result] The whole length of IL-2 gene of Sichuan white goose is 468 bp that contains a 441 bp open reading frame（ORF）,encoding 146 amino acid residues.Bioinformatics analysis shows that the amino acid sequence of IL-2 gene of Sichuan white goose contains four phosphorylation sites,a glycosylation site and a signal peptide with 21 amino acid residues.Homologies of IL-2 nucleotide sequence and amino acid sequence between Sichuan white goose and duck,chicken,turkey are 92.2%,77.5%,78.2% and 85.8%,65.5%,64.1%,respectively.By contrast IL-2 nucleotide sequence and amino acid sequence between Sichuan white goose and mammalian and rodents such as human,monkey,rat,bovine,horse,pig,cat,mouse,rabbit and deer,are all less than 45% and 28%,respectively.[Conclusion] The IL-2 gene of Sichuan white goose has closer genetic relationship with that of chicken and duck.