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鸡新城疫Ⅰ系疫苗诱导鸡胚IFNγ基因转录及cDNA克隆的研究
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张春杰， 程相朝， 李银聚， 吴庭才
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（河南科技大学 动物科技学院， 河南 洛阳 471003）
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摘要：
应用反转录-聚合酶链式反应（RT-PCR）技术，从鸡新城疫I系疫苗接种的洛阳土种鸡胚脾提取的RNA中扩增到干扰素γ（IFNγ）基因cDNA。结果表明，以1∶50或1∶100稀释度接种后第48h提取的mRNA扩增效果最佳。序列测定显示，所克隆的洛阳土种鸡IFNγ基因cDNA与所报道的鸡IFNγ基因cDNA同源性为99.7%，相应的氨基酸序列同源性为100%，说明鸡IFNγ基因编码区高度保守。 关键词：
洛阳土种鸡； 干扰素-γ； 转录； 克隆
中图分类号：S 852.42 文献标识码：
A 文章编号：1000-6419(2004)06-0012-04 |
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Transcription and cDNA cloning of IFNγ gene from
chicken embryo splenocytes induced by Newcastle disease virus
ZHANG Chun-jie, CHENG Xiang-chao， LI Yin-ju, WU Ting-cai
(College of Animal Science and Technology, Henan University of Science and Technology , Luoyang 471003,China) |
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Abstract: By reverse transcriptionpolymerase chain reaction
(RT-PCR), a cDNA encoding chicken interferongamma(chIFNγ) was cloned from Luoyang local chicken embryo splenocytes induced by
Newcastle disease virus Ⅰstrain. The result showed that the optimal time at which the mRNA of
IFNγ gene was extracted from spleen cells of chicken embryo is 48h after the embryo was inoculated by 1∶50 and 1∶100 diluted Newcastle disease virus. Sequence analysis showed that the
chIFNγcDNA is 492bp and encodes a protein including 164 amino acids. Compared with the reported nucleotide acid sequence of
IFNγ gene and the corresponding amino acid sequence of IFNγ, the nucleotide acid sequence of the
IFNγ gene cloned by us and the amino acid sequence deduced from the cloned
IFNγ gene shared 99.7% and 100% homology, respectively. It was concluded from the abovementioned results that the encoding region of
IFNγ gene is highly conservative.
Key words: Luoyang local chicken; interferongamma(IFNγ; transcription; cloning |
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