| 中国林蛙皮肤抗菌肽基因的cDNA克隆 |
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| 引用本文： | 乔红伟,韩俊友,赵瑞利,郑伟,韩艳,韩文瑜.中国林蛙皮肤抗菌肽基因的cDNA克隆[J].中国兽医学报,2006,26(6):646-648. |
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| 作者姓名： | 乔红伟 韩俊友 赵瑞利 郑伟 韩艳 韩文瑜 |
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| 作者单位： | 1. 吉林大学,畜牧兽医学院,吉林,长春,130062
2. 吉林省水产研究院,吉林,长春,130062 |
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| 基金项目： | 国家自然科学基金;吉林省科技厅应用基础研究项目 |
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| 摘 要： | 抗菌蛋白蛙皮抗菌肽，是两栖类动物中存在的天然免疫物质，在蛙类中仍然保持着一定的活性。我们用提取中国林蛙皮肤总RNA，逆转录合成cDNA，PCR扩增得到DNA片段，并与GenBank上登陆的其他蛙类的皮肤抗菌肽序列进行比较，发现此序列是一个新的蛙皮肤抗菌肽序列。PCR产物与pMD18-T连接，得到阳性重组体pMD18-RTCD。将阳性重组体进行测序，分析蛙皮素的氨基酸序列，显示它属于蛙皮抗菌肽家族。此外，中国林蛙抗菌肽氨基酸序列包含C-末端残基，同时具有两栖类动物抗菌肽的特征。这些信息对将来研究两栖类动物的天然免疫的分子机制具有重要作用。为应用基因工程技术对我国林蛙抗菌肽基因进行克隆表达奠定了基础。
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| 关 键 词： | 蛙皮抗菌肽基因 中国林蛙 PCR克隆 |
| 文章编号： | 1005-4545(2006)06-0646-03 |
| 收稿时间： | 2006-01-08 |
| 修稿时间： | 2006年1月8日 |
cDNA Cloning of the Antibacterial Peptide of the Rana Temporaria Chensinensis David Skin |
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| QIAO Hong-wei,HAN Jun-you,ZHAO Rui-li,ZHENG Wei,HAN Yan,HAN Wen-yu.cDNA Cloning of the Antibacterial Peptide of the Rana Temporaria Chensinensis David Skin[J].Chinese Journal of Veterinary Science,2006,26(6):646-648. |
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| Authors: | QIAO Hong-wei HAN Jun-you ZHAO Rui-li ZHENG Wei HAN Yan HAN Wen-yu |
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| Institution: | 1. College ofAnimal Science and Veterinary Medicine ,dilin University ,Changchun 130062 ,China;2. dilin Academy of Fishery Sciences ,Changchun 130033 ,China |
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| Abstract: | Antibacterial peptide of rana skin, an antibacterial protein, has been implicated in innate immunity m invertebrates. We cloned the Rana ternporaria chensinensis David skin cDNA using a PCR strategy and detected its activity in rana using a antibacterial protein assay against Micrococcus luteus. The cloning was based on a report of GenBank. The deduced amino acid sequence resulted in 53 amino with 46% identity to amphibia. RT-PCR was used to detect peptide mRNA in Rana temporaria chensinensis David. PCR products with T-4 linkagemse and pMD18-T vector. A positive recombinant clone pMD18-Cla obtained. Sequencing and anslysis of the amino acid sequence showed that the sequence contained extra residues at its C-type family of antibacterial peptide. Furthermore,the antibacterial peptide amino acid sequence contained extra residues at its C-terminus,which are characteristic of marine inverterates. This information will be useful in future studies on the molecular mechanisms of immunity in marine invertebrates,and provides the base for preparing the skin antibacterial peptide of the Rana temporaria chensinensis David with genetic engineering method. |
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| Keywords: | skin antibacterial peptide Rana ternporaria chensinensis David PCR clone |
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