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黑水虻新型抗菌肽基因的筛选、表达、纯化及功能鉴定

Screening, Expression, Purification and Functional Characterization of Novel Antimicrobial Peptide Genes from Hermetia illucens (L.).

作者信息

Elhag Osama, Zhou Dingzhong, Song Qi, Soomro Abdul Aziz, Cai Minmin, Zheng Longyu, Yu Ziniu, Zhang Jibin

机构信息

State Key Laboratory of Agricultural Microbiology, National Engineering Research Center of Microbial Pesticides, College of Life Science and Technology, Huazhong Agricultural University- Wuhan, China.

Faculty of Science and Technology, Omdurman Islamic University, Khartoum, Sudan.

出版信息

PLoS One. 2017 Jan 5;12(1):e0169582. doi: 10.1371/journal.pone.0169582. eCollection 2017.

Abstract

Antimicrobial peptides from a wide spectrum of insects possess potent microbicidal properties against microbial-related diseases. In this study, seven new gene fragments of three types of antimicrobial peptides were obtained from Hermetia illucens (L), and were named cecropinZ1, sarcotoxin1, sarcotoxin (2a), sarcotoxin (2b), sarcotoxin3, stomoxynZH1, and stomoxynZH1(a). Among these genes, a 189-basepair gene (stomoxynZH1) was cloned into the pET32a expression vector and expressed in the Escherichia coli as a fusion protein with thioredoxin. Results show that Trx-stomoxynZH1 exhibits diverse inhibitory activity on various pathogens, including Gram-positive bacterium Staphylococcus aureus, Gram-negative bacterium Escherichia coli, fungus Rhizoctonia solani Khün (rice)-10, and fungus Sclerotinia sclerotiorum (Lib.) de Bary-14. The minimum inhibitory concentration of Trx-stomoxynZH1 is higher against Gram-positive bacteria than against Gram-negative bacteria but similar between the fungal strains. These results indicate that H. illucens (L.) could provide a rich source for the discovery of novel antimicrobial peptides. Importantly, stomoxynZH1 displays a potential benefit in controlling antibiotic-resistant pathogens.

摘要

多种昆虫的抗菌肽对与微生物相关的疾病具有强大的杀菌特性。在本研究中,从黑水虻中获得了三种抗菌肽的七个新基因片段,分别命名为天蚕素Z1、杀菌肽1、杀菌肽(2a)、杀菌肽(2b)、杀菌肽3、嗜蝇肽ZH1和嗜蝇肽ZH1(a)。在这些基因中,一个189个碱基对的基因(嗜蝇肽ZH1)被克隆到pET32a表达载体中,并在大肠杆菌中作为与硫氧还蛋白的融合蛋白进行表达。结果表明,Trx-嗜蝇肽ZH1对多种病原体具有不同的抑制活性,包括革兰氏阳性菌金黄色葡萄球菌、革兰氏阴性菌大肠杆菌、真菌立枯丝核菌(水稻)-10和真菌核盘菌(Lib.)德巴利-14。Trx-嗜蝇肽ZH1对革兰氏阳性菌的最低抑菌浓度高于对革兰氏阴性菌,但在两种真菌菌株之间相似。这些结果表明,黑水虻可以为发现新型抗菌肽提供丰富的来源。重要的是,嗜蝇肽ZH1在控制耐药病原体方面显示出潜在的益处。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/401a/5215879/32aa744cb1b7/pone.0169582.g001.jpg

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