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北极狐白介素-18基因的克隆与序列分析
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张海玲,闫喜军*，罗国良，王凤雪，柴秀丽，易立，邵西群
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（中国农业科学院 特产研究所 人兽共患病研究室，吉林 吉林132109）
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摘要：
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根据GenBank中登录的犬白介素-18(IL-18)基因序列，设计了1对特异性引物。将北极狐外周血淋巴细胞经植物血凝素（PHA）诱导，用TRIzol裂解，提取总RNA，通过反转录聚合酶链式反应(RT-PCR)扩增狐狸IL-18基因，连接pMD18-T载体，转化DH5α感受态细胞，获得阳性重组质粒。核苷酸测序结果显示，获得的基因序列全长590bp，含582bp的开放阅读框，编码193个氨基酸，与已知犬IL-18序列的同源性高达98.7%，氨基酸同源性为96.4%。 |
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关键词：
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北极狐；白介素-18；克隆；序列分析 |
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中图分类号：Q 511：Q 503 文献标识码：
A 文章编号：1673-4696(2008)01-0059-04 |
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Cloning and sequence analysis of Alopex lagopus IL-18 gene |
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ZHANG Hai-ling,YAN Xi-jun,LUO Guo-liang,WANG Feng-xue,CHAI Xiu-li,YI Li,SHAO Xi-qun |
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(Department of Zoonoses,Institute of Special Wild Economic Animal and Plant Science,
Chinese Academy of Agricultural Sciences,Jilin 132109,China)
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Abstract: One pair of primers was designed and synthesized according to the dog IL-18 gene sequence in Genbank, and the IL-18 gene was amplified by RT-PCR from the total RNA of peripheral blood lymphocyte of Alopex lagopus induced with phytohemagglutinin (PHA). The RT-PCR product was cloned into the pMD18-T vector. Nucleotide sequence was determined after identification by digestion with restriction endonucleases.The results showed that the cloned Alopex lagopus IL-18 gene was 590bp in length, with a 582bp open reading frame encoding a polypeptide of 193 amino acids. The Alopex lagopus IL-18 gene shared 98.7% homology with the published dog IL-18 gene, and the deduced amino acid sequence from the IL-18 gene shared 96.4% homology with that from the dog IL-18 gene. |
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Key words:Alopex lagopus;interleukin-18;cloning;sequence analysis |
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