布鲁氏菌BP26蛋白对鼠源树突状细胞分化及抗原提呈作用  

Antigen presentation effect and differentiation of murine dendritic cells stimulated by Brucella BP26 protein

作　　者：徐龙 曹小安[2] 刘永生[2] 周建华[2] 张广林[1,2] 孙晶晶 李玲霞 赵永婕 胡永浩 XU Long;CAO Xiao-an;LIU Yong-sheng;ZHOU Jian-hua;ZHANG Guang-lin;SUN Jing-jing;LI Ling-xia;ZHAO Yong-jie;HU Yong-hao(College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China;State Key Laboratory of Veterinary Etiological Biology/Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730070, China)

机构地区：[1]甘肃农业大学动物医学院,甘肃兰州730070 [2]中国农业科学院兰州兽医研究所/家畜疫病病原生物学国家重点实验室,甘肃兰州730070

出　　处：《江苏农业学报》2020年第4期984-991,共8页Jiangsu Journal of Agricultural Sciences

基　　金：国家重点研发计划项目(2018YFD0500900)。

摘　　要：构建pET-30a-BP26并在大肠杆菌中表达获得重组布鲁氏菌BP26蛋白,研究重组表达蛋白对鼠髓源树突状细胞分化及抗原提呈作用。设计布鲁氏菌BP26蛋白引物,PCR扩增该基因并酶切、连接在pET-30a载体上。将构建成功的pET-30a-BP26转化至感受态细胞BL21表达获得重组BP26蛋白。采用SDS-PAGE和Western-Blot检测蛋白表达量和反应原性后,用重组蛋白刺激BALB/C小鼠分离髓源树突状细胞(BM-DCs),观察BM-DCs的成熟分化并利用流式细胞术进行表型分析。成功获得BP26重组蛋白,SDS-PAGE和Western-Blot分析结果表明,BP26重组蛋白可溶性高且具有良好的反应原性,流式细胞术检测结果显示,BP26能刺激BM-DCs分化成熟和抗原提呈作用。In this study,the pET-30a-BP26 was constructed and transfered into Escherichia coli to obtain recombinant protein BP26 of Brucella.In addition,the effects of recombinant protein on differentiation and antigen presentation of murine dendritic cells were studied.The Brucella BP26 protein specific primers were designed to amplify this target gene by PCR,then the gene was digested and linked to pET-30a vector.The pET-30a-BP26 was transformed into competent cell BL21,and the recombinant protein BP26 was dotained.The protein expression and reactogenicity were identified by SDS-PAGE and Western-blot.The bone marrow-derived dendritic cells(BM-DCs)were stimulated by recombinant protein.The differentiation of BM-DCs was observed,and the phenotype was analyzed by flow cytometry.The recombinant protein BP26 was successfully obtained.The results of SDS-PAGE and Western-blot showed that BP26 had high solubility and good reactogenicity.The results of flow cytometry showed that BP26 could simulate the differentiation,maturity and antigen presentation of BM-DCs.

关 键 词：布鲁氏菌 BP26 鼠源树突状细胞 抗原提呈 

分 类 号：Q816[生物学—生物工程]