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中国农学通报

中国农学通报 ›› 2011, Vol. 27 ›› Issue (14): 5-11.

所属专题： 生物技术； 畜牧兽医

• 畜牧 动物医学 蚕 蜂 • 上一篇    下一篇

鸡IL-2基因的克隆、序列分析及蛋白结构预测

谢昆   

  • 收稿日期:2010-06-28 修回日期:2010-08-26 出版日期:2011-06-15 发布日期:2011-06-15
  • 基金资助:

    国家科技支撑计划项目

Cloning and sequence analysis of gallus IL-2 gene and prediction of protein structure

  • Received:2010-06-28 Revised:2010-08-26 Online:2011-06-15 Published:2011-06-15

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摘要：

根据GenBank中已发表的鸡白细胞介素2 (IL-2)mRNA基因序列，利用Premier 5.0软件设计一对特异性引物，采用RT-PCR技术，以ConA刺激的鸡外周血淋巴细胞为材料，从总RNA中扩增出鸡IL-2基因。经琼脂糖凝胶电泳显示扩增片段约为432 bp，分离纯化片段，克隆入pMD-18T载体，转化DH5α感受态细胞，获得阳性重组质粒，经酶切鉴定，测序结果显示，我们得到了长432bp的基因，含有一个 432bp的开放阅读框，编码143个氨基酸。生物软件分析结果表明该序列与GenBank中发表的鸡的IL-2的核苷酸序列同源性为98.8%，与黄牛、马、鸡、绵羊、牛、犬、人、山羊、鼠、水牛的核苷酸同源性分别为31.2%、28.2%、27.3%、30.6%、26.2%、31.7%、30.1%、30.8%、26.6%、30.6%。与GenBank中发表的鸡IL-2编码的氨基酸序列同源性为95.8%，与上述动物的氨基酸序列同源性分别为7.6%、7.6%、9.7%、7.6%、6.9%、10.4%、11.8%、7.6%、9.0%、7.6%、10.4% 。这表明鸡的IL-2基因被成功克隆，它存在着种属特异性。这为进一步研究该基因的生物学作用特别是利用IL-2增强DNA疫苗的免疫效果奠定了基础。

关键词: 油棕 低温胁迫 光合作用 叶绿素荧光 能量耗散, 油棕 低温胁迫 光合作用 叶绿素荧光 能量耗散

Abstract:

According to GenBank in the IL-2 cDNA sequences, using Premier 5.0 software design a pairs of specific primer, the local Dorking inject 100 μg / ml ConA, total RNA was extracted from feeding 24 hours lymphocyte from spleen, by RT-PCR cloning colony IL-2 cDNA fragment. The DNA fragments were analyzed use DNAStar software between different species identity for comparison. Sequence analysis showed that IL-2 gene has an open reading frames for 432 nucleotide acids, encoding 143 amino acids and most of signal peptide, compared with published on Genbank, the nucleotide identity was 98.8%、31.2%、28.2%、27.3%、30.6%、26.2%、31.7%、30.1%、30.8%、26.6%、30.6%. compared with published on Genbank, the Amino acids was 95.8%, 7.6%、7.6%、9.7%、7.6%、6.9%、10.4% 11.8%, 7.6%, 9.0%, 7.6%, 10.4% with cattle, horse, cat, sheep, bovine, canis, people, capra, mice, cow of IL-2 sequence comparison The gallus IL-2 gene was successfully cloned,there is a species-specific. And construct a based on further study the biological effects of IL-2gene in particular the use of IL-2 enhance the immune effect of DNA vaccine.