牛蛙两个新Temporins基因的克隆及其抗菌机制的研究(英文)  

Molecular Cloning of Two Novel Temporins From Lithobates catesbeianus and Studying of Their Antimicrobial Mechanisms

作　　者：赵瑞利[1,2] 韩俊友[1] 韩文瑜[1] 雷连成[1] 孙长江[1] 冯新[1] 江丽娜[1] 乔红伟[3] 蔡林君[1] 

机构地区：[1]吉林大学畜牧兽医学院,长春130062 [2]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 [3]中国医学科学院实验动物研究所,北京100021

出　　处：《生物化学与生物物理进展》2009年第8期1064-1070,共7页Progress In Biochemistry and Biophysics

基　　金：supported by a grant from The National Natural Science Foundation of China (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250mg/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≈230μmol/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.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（LLRHVVKILEKYLamide） and temporin-Lb（LFRHVVKIFEKYLamide） 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 Stap hylococc us aure us.

关 键 词：抗菌肽 牛蛙 temporins 透射电镜 

分 类 号：Q5[生物学—生物化学]