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| 牛蛙两个新Temporins基因的克隆及其抗菌机制的研究 | |
| 引用本文： | 赵瑞利,韩俊友,韩文瑜,雷连成,孙长江,冯 新,江丽娜,乔红伟,蔡林君. 牛蛙两个新Temporins基因的克隆及其抗菌机制的研究[J]. 生物化学与生物物理进展, 2009, 36(8): 1064-1070. DOI: 10.3724/SP.J.1206.2009.00033 |
| 作者姓名： | 赵瑞利 韩俊友 韩文瑜 雷连成 孙长江 冯 新 江丽娜 乔红伟 蔡林君 |
| 作者单位： | 1. 吉林大学畜牧兽医学院,长春,130062;中国科学院动物研究所,动物生态与保护生物学院重点实验室,国家野生动物疫病研究中心,北京,100101 2. 吉林大学畜牧兽医学院,长春,130062 3. 中国医学科学院实验动物研究所,北京,100021 |
| 基金项目： | 国家自然科学基金资助项目(30571416) |
| 摘 要： | Temporins 是从蛙属中得到的一类羧基端酰胺化的疏水性抗菌肽，具有抗细菌、霉菌、酵母菌、原虫及病毒活性．为了研究牛蛙皮肤抗菌肽的多样性及其结构特点，根据GenBank数据库中蛙属抗菌肽基因信号肽序列设计简并引物，从牛蛙皮肤cDNA文库中克隆到两个新的temporins 家族抗菌肽，命名为 temporin-La (LLRHVVKILEKYLamide) 和 temporin-Lb (LFRHVVKIFEKYLamide)．合成的 temporin-La 和 temporin-Lb肽具有很强的抗菌活性，尤其是对革兰氏阳性细菌．溶血性测定结果表明，temporin-La 浓度高至250 mg/L 时对兔红细胞仍无溶血活性，而 temporin-Lb 具有较弱的溶血活性(半数致死浓度LC50 ≈ 230 μmol/L)．通过透射电镜观察 temporin-La 和 temporin-Lb 处理过的金黄色葡萄球菌的细胞结构，发现它们都能直接地杀死细菌，但作用机制不一样． |
| 关 键 词： | 抗菌肽，牛蛙，temporins，透射电镜 |
| 收稿时间： | 2009-01-13 |
| 修稿时间： | 2009-05-11 |
Molecular cloning of two novel temporins from Lithobates catesbeianus and studying of their antimicrobial mechanisms |
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| ZHAO Rui-Li,HAN Jun-You,HAN Wen-Yu,LEI Lian-Cheng,SUN Chang-Jiang,FENG Xin,JIANG Li-N,QIAO Hong-Wei and CAI Lin-Jun. Molecular cloning of two novel temporins from Lithobates catesbeianus and studying of their antimicrobial mechanisms[J]. Progress In Biochemistry and Biophysics, 2009, 36(8): 1064-1070. DOI: 10.3724/SP.J.1206.2009.00033 | |
| Authors: | ZHAO Rui-Li HAN Jun-You HAN Wen-Yu LEI Lian-Cheng SUN Chang-Jiang FENG Xin JIANG Li-N QIAO Hong-Wei CAI Lin-Jun |
| Affiliation: | College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China; National Research Center for Wildlife Born Diseases, Key Laboratory of Animal Ecology and Conservation Biology,Institute of Zoology, The Chinese Academy of Sciences, Beijing 100101, China;College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China;College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China;College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China;College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China;College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China;College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China;Institute of Laboratory Animal Sciences, Chinese Academy of Medical Sciences(CAMS) & Peking Union Medical College(PUMC), Beijing 100021,China;College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China |
| Abstract: | Temporins are a kind of small,hydrophobic and C-terminus amidated antimicrobial peptides from Rana species.They are effective against bacteria,fungi,yeast,protozoa and viruses.Two novel temporins named as temporin-La(LLRHWKILEKYLanifc) and temporin-Lb(LFRHVVKIFEK.Ylamid.) were cloned from Lithobates catesbeianus.Synthetic peptides of temporin-La and temporin-Lb showed strong antimicrobial activities against bacteria tested,especially Gram-positive bacteria.Besides,temporin-La showed no haemolytic activity to rabbit erythrocytes at the concentration of 250 mg/L while temporin-Lb showed weak haemolytic activity(LC50≈ 230 μmol/L).Transmission electron microscopy showed that temporin-La and temporin-Lb induced different effects on bacterial structure of Staphylococcus aureus. |
| Keywords: | antimicrobial peptides Lithobates catesbeianus temporins transmission electron microscopy |
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