摘要
为了在在大肠杆菌中融合表达杂合抗菌肽BM16并研究其生物学活性,本试验利用生物信息学分析软件设计出1条杂合抗菌肽BM16,运用同源重组的方法构建重组表达质粒p ET-28a（＋）-Ub-BM16,在大肠杆菌BL21（DE3）中诱导表达,采用Ni2＋亲和层析纯化获得目的蛋白。结果显示,表达出13.2 ku的Ub-BM16融合蛋白,经SUMO酶切后获得2.0 ku的目的蛋白,纯化后纯度大于95%,该多肽对大肠杆菌、沙门菌和金黄色葡萄球菌等均有抑制作用。上述研究结果为开发新型抗菌肽提供了思路。
The study aims to investigate the fusion expression system and biological activity of the hybrid antimicrobial peptide BM16 expressed from E.coli.The hybrid antimicrobial peptide BM16 was designed by bioinformatics analysis software and the recombinant expression plasmid p ET28 a-Ub-BM16 was constructed using the homologous recombination method.The recombinant plasmid was induced to express in E.coli BL 21（DE3） and the recombinant protein was successfully expressed and purified using Ni2＋affinity chromatography,which was 13.2 ku.The target protein of 3.0 ku was obtained by SUMO enzyme digestion of the recombinant protein.The purity of the target protein is higher than 95%.The polypeptide protein exhibited an inhibitory effect on Escherichia coli,Salmonella and Staphylococcus aureus.The above-mentioned results provide an idea for the development of new antimicrobial peptides.
作者
周婧然
刘磊
林鑫
孔令聪
裴志花
刘树明
马红霞
ZHOU Jing-ran;LIU Lei;LIN Xin;KONG Ling-cong;PEI Zhi-hua;LIU Shu-ming;MA Hong-xia(a. College of A nimal A cience and Technolog;b. Animal Production＆Product Quality and Security of the Ministry of Education, Jilin Agricultural University, Changchun130118 , Chin)
出处
《中国兽医科学》
CAS
CSCD
北大核心
2018年第7期899-904,共6页
Chinese Veterinary Science
基金
国家自然科学基金资助项目(31572574,31502121)
国家重点研发专项计划项目(2016YFD0501301,2016YFD0501401)
吉林省科技发展计划项目(20160204002NY)
关键词
杂合抗菌肽
BM16
融合表达
抑菌活性
hybrid antimicrobial peptide
BM16
fusion expression
bacteriostatic activity
