动物营养学报2016，Vol.28Issue(4)：1114-1121,8.DOI:10.3969/j.issn.1006-267x.2016.04.020
重组鹅β-防御素7蛋白的原核表达及其生物学特性分析
Recombinant Expression and Biological Property Analysis of Goose Avian β-Defensin 7 Protein
摘要
Abstract
The objective of the study was to investigate the biological property of goose avian β-defensin ( AvBD) 7. The gene of AvBD7 was cloned into EcoRⅠ and XhoⅠ sites of pProEX HTa vector to construct recombinant plasmid pProEX-AvBD7. The recombinant plasmid was transformed into Escherichia coli Rosetta and the bacteria was induced with isopropy-β-D-thiogalactoside ( IPTG) . It was analyzed by tricine sodium do-decyl sulfate polyacrylamide gel electrophoresis ( Tricine-SDS-PAGE) that the recombinant protein size was 10 to 15 ku, which was the same as the expected size. Antimicrobial activity of the recombinant fusion protein a-gainst Escherichia coli, Salmonella pullorum, Staphylococcus aureus, Micrococcus tetragenus and Bacillus subtilis was measured in vitro. In addition, the effect of ionic strength on the antibacterial activity and hemolytic activity of the recombinant fusion protein were investigated. The results showed that the recombinant goose AvBD7 protein exhibited significant antimicrobial activity against the five bacteria investigated ( P<0.05) , and antibacterial activity enhanced with the increasing of protein concentration. The antibacterial activity of recombi-nant goose AvBD7 protein was significantly inhibited by high salt concentration (150 mmol/L) (P<0.05). In addition, recombinant goose AvBD7 protein showed little hemolytic activity against chicken erythrocytes ( P>0.05). In conclusion, recombinant goose AvBD7 protein exhibits extensive antimicrobial activity. High salt concentration significantly decrease its antibacterial activity. In addition, the recombinant goose AvBD7 protein shows little hemolytic activity.关键词
鹅β-防御素7/重组蛋白/抗菌活性Key words
goose β-defensin 7/recombinant protein/antimicrobial activity分类
农业科技引用本文复制引用
高梦莹,徐倩倩,张婷婷,陈玉秋,赵文钧,齐甜铭,马得莹..重组鹅β-防御素7蛋白的原核表达及其生物学特性分析[J].动物营养学报,2016,28(4):1114-1121,8.基金项目
教育部博士点基金项目(20122325110016) （20122325110016）
哈尔滨市科技创新人才研究专项资金项目(2013RFXXJ019) （2013RFXXJ019）
黑龙江省应用技术研究与开发计划项目( PC13S02) （ PC13S02）
