重组鸡β-防御素2蛋白的表达与生物学活性鉴定

为探讨鸡β-防御素2（Av BD2）的生物学特性,研究将Av BD2基因克隆到原核表达载体p Pro EX HTa的Eco RⅠ和XhoⅠ双酶切位点上,构建重组表达质粒p Pro EX-Av BD2,将重组质粒转化大肠杆菌Rosetta感受态细胞,用IPTG对其菌液进行诱导表达。Tricine-SDS-PAGE电泳结果表明,His-Av BD2重组蛋白分子质量约12 ku,与预期一致。将该重组蛋白纯化后,通过菌落计数法测定其体外抗肠炎沙门氏菌、四联球菌、大肠杆菌和枯草芽孢杆菌等抑菌活性,盐离子浓度对其抗菌活性的影响及其溶血活性。结果显示,Av BD2重组蛋白对所测定的4株细菌有显著抗菌活性,高...

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Published in东北农业大学学报 Vol. 46; no. 8; pp. 56 - 60
Main Author 马得莹 张婷婷 徐杨 高梦莹 王秋玲 徐倩倩 韩宗玺 刘胜旺
Format Journal Article
LanguageChinese
Published 中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室禽传染病研究室,哈尔滨150001%中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室禽传染病研究室,哈尔滨,150001 2015
东北农业大学动物科学技术学院,哈尔滨,150030%东北农业大学动物科学技术学院,哈尔滨150030
Subjects
AvBD2
抗菌活性
重组蛋白
鸡
鸡
AvBD2
chicken
recombinant protein
β-defensin 2
重组蛋白
antimicrobial activity
抗菌活性
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Summary:为探讨鸡β-防御素2（Av BD2）的生物学特性,研究将Av BD2基因克隆到原核表达载体p Pro EX HTa的Eco RⅠ和XhoⅠ双酶切位点上,构建重组表达质粒p Pro EX-Av BD2,将重组质粒转化大肠杆菌Rosetta感受态细胞,用IPTG对其菌液进行诱导表达。Tricine-SDS-PAGE电泳结果表明,His-Av BD2重组蛋白分子质量约12 ku,与预期一致。将该重组蛋白纯化后,通过菌落计数法测定其体外抗肠炎沙门氏菌、四联球菌、大肠杆菌和枯草芽孢杆菌等抑菌活性,盐离子浓度对其抗菌活性的影响及其溶血活性。结果显示,Av BD2重组蛋白对所测定的4株细菌有显著抗菌活性,高盐浓度显著降低其抗菌活性。该重组蛋白溶血活性极低。
Bibliography:23-1391/S
MA Deying, ZHANG Tingting, XU Yang, GAO Mengying, WANG Qiuling, XU Qianqian, HAN Zongxiz, LIU Shengwang（1. School of Animal Science and Technology, Northeast Agricultural University, Harbin 150030, China; 2. Division of Avian Infectious Disease, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China）
The objective of the study was to investigate bioactivity of chicken avian β-defensin （AvBD） 2 protein. The cDNA of AvBD2 was cloned into EcoR I and Xho I sites of pProEX HTa vector to construct recombinant plasmid pProEX-AvBD2. The recombinant piasmid was transformed into E. coli Rosetta and the bacteria was induced with IPTG. It was demonstrated by Tricine-SDS-PAGE that a 12 kD protein was highly expressed. Antimicrobial activity of the recombinant fusion protein against Salmonella enteritidis, Bacillus subtilis, Micrococcus tetragenus, and E. coll. was measured in vitro. In addition, effect of ionic stren
ISSN:1005-9369