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广西10个地方优质品种鸡γ-干扰素基因
的克隆与序列分析
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黄溢泓1，谢芝勋2，孙建华2
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（1.广西大学 动物科学技术学院，广西 南宁530005； 2.广西兽医研究所，广西 南宁530001）
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摘要：
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根据基因库中鸡γ-干扰素的基因序列设计了1对特异性引物，应用反转录-聚合酶链式反应(RT-PCR)技术，从ConA诱导培养的广西10个地方优质品种鸡外周血淋巴细胞RNA中扩增了γ-干扰素基因，结果，均得到了大小为520bp的特异性片段。将扩增产物纯化并克隆到pMD18-T载体上，获得重组质粒，经PCR和EcoRⅠ+SalⅠ双酶切鉴定后测序。序列分析结果表明，广西10个地方优质品种鸡γ-干扰素基因均编码145个氨基酸的成熟蛋白，分子质量约为16.8ku，与哺乳动物和其他品种鸡的核苷酸序列同源性分别为38.8%～39.0%和99.2%～100%，氨基酸序列同源性分别为236%～248%和97.6%～99.4%。 |
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关键词：
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γ-干扰素基因；鸡；克隆；序列分析 |
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中图分类号：S 859.797：Q 785 文献标识码：
A 文章编号：1673-4696(2006)05-0415-05 |
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Molecular cloning and sequence analysis of interferon gamma gene
from ten local breeds of chickens in Guangxi |
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HUANG Yi-hong1，XIE Zhi-xun2，SUN
Jian-hua2 |
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（1. College of Animal Science and Technology, Guangxi University, Nanning 530005,China；
2. Guangxi Veterinary Research Institute, Nanning 530001,China） |
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Abstract: A pair of specific primers was designed according to the sequence of chicken interferon-gamma gene (ChIFN-γ) published in GenBank. The ChIFN-γ gene was amplified by RT-PCR from chickens’ spleen lymphocytes of ten local breeds, which were activated by concanavalin A(ConA). After being cloned into the pMD18-T vector, the nucleotide acid sequences of ChIFN-γ gene were identified by PCR and digestion with restrictive endonucleases. Results showed that the coding region of ChIFN-γ gene encoded 145 amino acids, with a predicted mature protein of 16.8ku. Compared with that of mammals and other chickens, the sequence homology were 38.8%-39.0% and 99.2%-100% in nucleotide acids, respectively, and 23.6%-24.8% and 97.6%-99.4% in amino acids, respectively. |
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Key words: interferon-gamma gene; chicken; cloning; sequence analysis |
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