中文摘要：

目的建立布鲁菌免疫蛋白质组学方法,从羊布鲁菌膜蛋白中筛选免疫候选抗原。方法提取羊布鲁菌16M膜蛋白,进行双向电泳（2-DE）分离,分别用牛、羊布鲁菌感染的阳性血清进行Westernblot分析,筛选出的免疫显性蛋白点进行LC-MS/MS质谱鉴定,并进行生物信息学分析。结果筛选出56个免疫显性蛋白点,鉴定成功35个,其中有12个蛋白是能够被牛、羊2种血清共同识别的抗原。数据检索显示,这些蛋白主要涉及布鲁菌的能量代谢,蛋白质、氨基酸合成,脂肪酸代谢及糖和辅酶的合成等生物过程。结论利用免疫蛋白质组学技术成功筛选出羊布鲁菌高免疫原性膜蛋白,为布鲁菌亚单位疫苗的研制提供了大量的候选抗原。

英文摘要：

Objective To develop a immunoproteomical method for screening of candidate antigen for immunization from membrane protein of Brucella melitensis.Methods The membrane protein of Brucella melitensis 16M was extracted,then separated by two-dimensional electrophoresis（2-DE）and subjected to Western blot with human,bovine and sheep sera infected with brucella.The screened immunodominant protein dots were identified by LC-MS/MS and analyzed for bioinformatics.Results A total of 56 immunodominant protein dots representing 35 open reading frames（ORFs）were screened,of which 12 were co-recognized by bovine and sheep sera.Bioinformatics retrieval showed that the proteins were mainly associated with the energy metabolism,protein and amino acid syntheses,fatty acid metabolism as well as saccharide and coenzyme syntheses of brucella.Conclusion High immunogenic membrane protein was successfully screened from Brucella melitensis by immunoproteomics,which provided a large quantity of candidate antigens for preparation of brucella subunit vaccine.