重组鹅β-防御素12蛋白的原核表达及生物学特性

发布时间：2018-05-19 21:06

本文选题：鹅β-防御素 + 重组蛋白　；参考：《微生物学通报》2016年09期

【摘要】：【目的】研究重组鹅β-防御素12蛋白的原核表达并探究其生物学特性。【方法】采用His标签蛋白原核表达系统,将鹅防御素12(Av BD12)基因亚克隆到表达载体p Pro EX-HTa上,构建重组表达质粒。将重组表达质粒转化到大肠杆菌Rosseta感受态中,用IPTG进行诱导表达,并对该重组蛋白进行纯化。进一步采用菌落计数法测定其体外抗菌活性和盐离子稳定性。【结果】经Tricine-SDS-PAGE电泳分析,诱导表达的鹅Av BD12重组蛋白分子量约为12 k D,大部分以包涵体形式存在。该重组蛋白对大肠杆菌、鸡白痢沙门氏菌、金黄色葡萄球菌、四联球菌、枯草芽孢杆菌均具有抗菌活性,高浓度盐离子显著抑制重组蛋白的抗菌活性。此外,该重组蛋白对鸡红细胞没有溶血活性。【结论】该重组蛋白具有广谱抗菌活性,高浓度盐离子显著降低其抗菌活性,且该重组蛋白不具有溶解鸡红细胞的活性。
[Abstract]:[objective] to study the prokaryotic expression of recombinant goose 尾 -defensin 12 protein and explore its biological characteristics. [methods] 12(Av BD12 gene was subcloned into the expression vector p Pro EX-HTa by using His tag protein prokaryotic expression system. The recombinant expression plasmid was constructed. The recombinant expression plasmid was transformed into Escherichia coli (E. coli) Rosseta competent state and was induced to express by IPTG, and the recombinant protein was purified. The antibacterial activity and salt ion stability in vitro were determined by colony counting method. [results] by Tricine-SDS-PAGE electrophoresis, the molecular weight of the recombinant protein induced by Av BD12 was about 12 KD, most of which were in the form of inclusion body. The recombinant protein had antibacterial activity against Escherichia coli, Salmonella pullorum, Staphylococcus aureus, Tetracoccus tetralogus and Bacillus subtilis. In addition, the recombinant protein has no hemolytic activity on chicken red blood cells. [conclusion] the recombinant protein has broad-spectrum antibacterial activity, the high concentration of salt ions significantly reduces its antibacterial activity, and the recombinant protein does not have the activity of dissolving chicken red blood cells.
【作者单位】：东北农业大学动物科技学院;
【基金】：教育部博士点基金项目(No.20122325110016) 哈尔滨市科技创新人才研究专项资金项目(No.2013RFXXJ019) 黑龙江省应用技术研究与开发计划项目(No.PC13S02)~~
【分类号】：S852.2

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