效应蛋白在布鲁氏菌毒力和免疫保护性中的作用机制  

Mechanism of effector proteins in virulence and immunoprotection in Brucella

作　　者：吴海燕[1] 丁洁蔚 杨朝晖[1] 沈扬[1] WU Haiyan;DING Jiewei;YANG Chaohui(Department of Infectious Diseases,Taizhou First People’s Hospital,Taizhou 318020,China)

机构地区：[1]台州市第一人民医院感染科,浙江台州318020 [2]浙江省台州医院老年医学科

出　　处：《全科医学临床与教育》2023年第7期584-587,F0002,共5页Clinical Education of General Practice

基　　金：浙江省医药卫生科技计划(2023KY398)。

摘　　要：目的分析效应蛋白在布鲁氏菌毒力和免疫保护性中的作用机制。方法以布鲁氏菌为模板,采用PCR扩增BspC基因,后进行测序,观察布鲁氏菌的基因组学、布鲁氏菌属细菌中BspC蛋白,以及BspC蛋白是否存在免疫原性,BspC蛋白的三维结构建模,BspC蛋白的克隆、表达、纯化,BspC蛋白的免疫胶体金实验。结果B29基因组大小为3.3 Mb,GC%含量57.2%,有两条环状染色体,大小分别2126260 bp、1185609 bp,含有3368个编码基因,64个RNAs。布鲁氏菌属存在于各个种之间,且相对保守,均显示为100%相似度,未出现突变。BspC蛋白的N段存在明显的信号肽特征,在第20、23和25位的氨基酸有较为显著的预测值,BspC蛋白N端1-23位氨基酸预测为胞内和跨膜区域,BspC蛋白为布鲁氏菌特有的蛋白。BspC蛋白并粗纯化后蛋白分子量<13.9 kD。显微镜下检测结果显示,细菌的细胞膜外周可出现黑褐色颗粒,且在细胞质中也显示颗粒。结论重组蛋白BspC具有免疫反应原性;BspC可能在布鲁氏菌感染宿主过程中发挥免疫逃逸的作用。Objective To analyze the mechanisms of effector proteins in Brucella virulence and immunoprotection.Methods BspC gene was amplified by PCR using Brucella as template and sequenced to observe the genomics of Brucella,immunogenicity of BspC protein,3D structure modeling of BspC protein,the cloning,expression,and purification of BspC protein,immunocolloidal gold experiments of BspC protein.Results The B29 genome is 3.3 Mb with 57.2%of GC%and two circular chromosomes which are 2126260 bp and 1185609 bp,containing 3368 coding genes and 64 RNAs.Brucella genus exist in every species,and relatively conserved,all show 100%identity,no mutation.The N segment of BspC protein has obvious signal peptide characteristics,with significant predictive values at amino acids 20,23 and 25,amino acids 1-23 at the N terminus of BspC protein are predicted to be intracellular and transmembrane regions,and BspC protein is unique to Brucella.BspC protein and crude purified protein molecular weights were<13.9 kD.Microscopic detection results showed that black-brown granules could appear in the periphery of the cell membrane of the bacteria,and the granules could be displayed in the cytoplasm.Conclusion BspC is immunoreactive.BspC may play a role in immune escape during Brucella host infection.

关 键 词：效应蛋白 布鲁氏菌 毒力 免疫保护 

分 类 号：R392[医药卫生—免疫学]