动物医学进展2024，Vol.45Issue(9)：39-44,6.
牛种布鲁氏菌ArsR2蛋白的原核表达及磷酸化位点鉴定
Prokaryotic Expression and Phosphorylation Site Identification of ArsR2 Protein of Brucella abortus
摘要
Abstract
The purpose of this study was to express and purify the ArsR2 protein of Brucella abortus S2308 strain by prokaryotic expression system,analyze the related information of the protein by bioinformatics software,and identify the modification site of the protein by mass spectrometry.The arssR2 gene was ampli-fied by PCR using the genome of Brucella abortus S2308 strain as a template,and the pET-32a-ArsR2 re-combinant plasmid was constructed.After sequencing,the recombinant plasmid was transformed into E.co-li competent cells BL21(DE3)for expression under induction of IPTG.The ArsR2 protein was purified by Ni-NTA affinity chromatography and identified by SDS-PAGE and Western blot.The physical and chemical properties and phosphorylation sites of ArsR2 protein of Brucella abortus were predicted by bioinformatics methods.On this basis,the modification sites of ArsR2 protein were identified by mass spec-trometry.The results showed that ArsR2 protein was successfully expressed and purified.Through bioin-formatics analysis,it was found that ArsR2 protein had no transmembrane region,and phosphorylation modification.The ArsR2 protein was further identified by mass spectrometry to have two phosphorylation sites.The results showed that the ArsR2 protein of Brucella abortus S2308 strain expressed in prokaryotic expression had phosphorylation modification sites,which laid a foundation for further exploring the biologi-cal function of the protein.关键词
布鲁氏菌/arsR2基因/原核表达/蛋白纯化Key words
Brucella/arsR2 gene/prokaryotic expression/protein purification分类
农业科技引用本文复制引用
刘科蒙,耿好,苏梦茹,许健,支飞杰,储岳峰..牛种布鲁氏菌ArsR2蛋白的原核表达及磷酸化位点鉴定[J].动物医学进展,2024,45(9):39-44,6.基金项目
中国博士后基金项目(2022M721437) （2022M721437）
甘肃省自然科学基金重点项目(23JRRA560) （23JRRA560）
甘肃省科技重大专项课题(22ZD6NA001-04) （22ZD6NA001-04）
甘肃省科研联合基金(23JRRA1517) （23JRRA1517）
国家自然基金面上项目(32373017) （32373017）
