thanatin串联抗菌肽原核表达研究  

Expression of antimicrobial peptides thanatin tandem fusion in Escherichia coli

作　　者：刘伟[1] 李清海[2] 刘仁虎[3] LIU Wei1,LI Qinghai2,LIU Renhu3(1 .Institute of Plant Protection and Microbiology,Zhejiang Academy of Agricultural Sciences,Hangzhou 310021,China ; 2. Department ooRadiology,The Second Affilidted Hospital of Zhejiang University School of Medicine,Hangzhou 310009, China; 3. Institute Virologyand Biotechnology,Zhejiang Acadeey of Agricultural Sciences,Hangzhou 310021 , Chin)

机构地区：[1]浙江省农业科学院植物保护与微生物研究所,浙江杭州310021 [2]浙江大学医学院附属第二医院放射科,浙江杭州310009 [3]浙江省农业科学院病毒生物技术研究所,浙江杭州310021

出　　处：《浙江农业学报》2018年第5期804-809,共6页Acta Agriculturae Zhejiangensis

基　　金：浙江省植物有害生物防控重点实验室-省部共建国家重点实验室培育基地(2010DS700124-ZZ1604;2010DS700124-ZZ1701);浙江省自然科学基金(LY15H180004);浙江省公益项目(2015C32020)

摘　　要：抗菌肽thanatin是一类广谱性的阳离子抗菌活性肽,对病原性真菌有很好的杀灭效果。为了提高thanatin抗菌肽表达丰度,将3个thanatin单体拷贝基因进行串联,与MBP蛋白进行融合表达,表达的产物经过酶切纯化后,通过Western blot和质谱鉴定,结果表明3×thanatin在大肠埃希菌中成功表达,对核盘菌的抑菌活性测定结果表明,原核表达的3×thanatin最低抑菌浓度为6.6μmol·L^(-1),而化学合成的thanatin抗菌肽对核盘菌的抑菌浓度约为64μmol·L^(-1),3×thanatin抗菌肽对核盘菌的抑菌活性提升了约1个数量级。The antibacterial peptide thanatin is a broad spectrum cationic antibacterial peptide,which can significantly kill pathogenic fungi.In order to improve the expression level of thanatin,three copies of thanatin monomer and MBP protein was co-expressed in Escherichia coli.The expression products were identified by Western blot and mass spectrometry after enzyme digestion and purification.The results showed that the three copies of thanatin fusion protein,3×thanatin,were successfully expressed in E.coli and had good inhibition activity on Sclerotinia sclerotiorum.The minimum concentration of inhibition on S.sclerotiorum was 6.6μmol·L-1for 3×thanatin,which was about ten times higher than 1×thanatin.

关 键 词：抗菌肽 THANATIN 串联 表达 

分 类 号：S432.23[农业科学—植物病理学]