thanatin 串联抗菌肽原核表达研究

doi:10.3969/j.issn.1004-1524.2018.05.18

浙江农业学报 ›› 2018, Vol. 30 ›› Issue (5): 804-809.DOI: 10.3969/j.issn.1004-1524.2018.05.18

thanatin 串联抗菌肽原核表达研究

刘伟¹, 李清海², 刘仁虎^3,*

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

收稿日期:2017-09-07 出版日期:2018-05-20 发布日期:2018-05-23
通讯作者: 刘仁虎,E-mail: liurh@mail.zaas.ac.cn
作者简介:刘伟(1978—),男,河南信阳人,主要从事抗菌肽相关研究。E-mail: biolwei@sina.com
基金资助:
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)

Expression of antimicrobial peptides thanatin tandem fusion in Escherichia coli

LIU Wei¹, LI Qinghai², LIU Renhu^3,*

1.Institute of Plant Protection and Microbiology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China;
2. Department of Radiology, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou 310009, China;
3. Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China

Received:2017-09-07 Online:2018-05-20 Published:2018-05-23

摘要/Abstract

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6.6 μmol·L^-1, 而化学合成的thanatin抗菌肽对核盘菌的抑菌浓度约为64 μmol·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。

关键词: 抗菌肽, thanatin, 串联, 表达

Abstract: 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 6.6 μmol·L^-1 for 3×thanatin, which was about ten times higher than 1×thanatin.

Key words: antimicrobial peptides, thanatin, tandem, expression

中图分类号:

S432.2⁺3

刘伟, 李清海, 刘仁虎. thanatin 串联抗菌肽原核表达研究[J]. 浙江农业学报, 2018, 30(5): 804-809.

LIU Wei, LI Qinghai, LIU Renhu. Expression of antimicrobial peptides thanatin tandem fusion in Escherichia coli[J]. , 2018, 30(5): 804-809.

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thanatin 串联抗菌肽原核表达研究

Expression of antimicrobial peptides thanatin tandem fusion in Escherichia coli

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