布鲁菌BMEI1829基因的原核表达、蛋白纯化及动物免疫试验  

Procaryotic Expression of Brucella BMEI1829 Gene,Protein Purification and Animal Immunization Test

作　　者：赵晓燕[1,2] 朱亮[3] 姜德玉[2] 高啸[2] 段越强[2] 王希良[2] 李培锋[1] 

机构地区：[1]内蒙古农业大学动物医学与科学学院,内蒙古呼和浩特010018 [2]军事医学科学院微生物流行病研究所病原微生物生物安全国家重点实验室,北京100071 [3]中国农业大学农学与生物技术学院,北京100193

出　　处：《动物医学进展》2010年第10期5-9,共5页Progress In Veterinary Medicine

基　　金：国家973计划资助项目(2010CB530202)

摘　　要：利用PSORTb及CELLO生物学软件对布鲁菌16M菌株基因组序列进行分析,预测其外膜蛋白基因序列;选取BMEI1829基因进行PCR扩增并与原核表达载体pET32a(+)连接,转入E.coliBL21(DE3)感受态细胞;IPTG诱导蛋白表达,SDS-PAGE分析及Western blot鉴定目的蛋白的表达;经His-TrapTMHP纯化,免疫Balb/c小鼠,间接ELISA法检测特异性抗体。结果表明,BMEI1829蛋白在原核表达系统中成功表达,纯化后蛋白免疫Balb/c小鼠可产生特异性抗体,抗体亚型分布以IgG1、IgG2b为主,为进一步研究其功能奠定了基础。Brucella strain 16M genome sequence was analyzed by PSORTb and CELLO to predict the genetic sequence of outer membrane proteins;BMEI1829 gene was selected to amplify by polymerase chain reactions.The PCR product was cloned into the prokaryotic expression vector pET32a（＋）,which was then transfected into the competent cells E.coli BL21（DE3）.The protein expression was induced by IPTG and examined with SDS-PAGE and Western blot,and purified by HisTrapTM HP,then Balb/c mice were immunized with the purified protein,indirect ELISA was used to detect specific antibody.BMEI1829 gene was successfully expressed in the prokaryotic expression system,The target protein was purified and used to immunize Balb/c mice,which produced specific antibodies that mainly consist of the subtypes IgG1 and IgG2b.This study lays a foundation for further study on the function of protein.

关 键 词：布鲁菌 BMEI1829基因 原核表达 

分 类 号：S852.614[农业科学—基础兽医学]