运用免疫蛋白质组学方法筛选副猪嗜血杆菌血清4型菌株的免疫反应性蛋白

Immunoreactive proteins of Haemophilus parasuis serotype 4 isolates by an immunoproteomic assay

选取2株副猪嗜血杆菌血清4型菌株(A、B菌株),采用蛋白提纯和双向电泳技术获得高分辨率的蛋白胶。通过比对蛋白胶和免疫印迹膜上的蛋白反应点,并切取相应蛋白点进行质谱鉴定。根据质谱鉴定结果,菌株A和B有13个相同的蛋白点,其中8个蛋白点是新发现的具有免疫原性的蛋白点,它们分别是二氢硫辛酰胺脱氢酶(DLDH)、dppA、过氧化氢酶(CAT)、蛋白酶(P)、转酮醇酶(TK)、延长因子(EF-Ts)、周质丝氨酸蛋白水解酶(SP)、2,3-二磷酸甘油酸变位酶(BPGM)。在2株副猪嗜血杆菌血清4型菌株中发现有13个相同的免疫反应性蛋白,DLDH、dppA、CAT、P、TK、EF-Ts、SP和BPGM是新发现的免疫原性蛋白,它们作为疫苗候选抗原有待进一步研究。

To search the immunogenic proteins of H. parasuis strains serotype 4 in order to provide the novel vaccine candidate antigens. A protein extraction method for two H. parasuis strains serotype 4（strain A and strain B） and two-dimensional electrophoresis（2-DE） were optimized to provid high-resolution profiles. Spots were excised from gels after strict comparison with 2-DE Western blot membranes and relevant gels,and indentified by mass spectrometry. Compared to mass spectrum results,13 proteins were identified to be the same between strain A and strain B. Eight of them were reported as immunoreactive proteins in H. parasuis for the first time. They are dihydrolipoamide dehydrogenase （ DLDH ）, dppA, catalase （ CAT）, protease （ P）, transketolase （ TK）, elongation factor Ts（EF-Ts）,periplasmic serine protease do/hhoA-like protein（SP）,and 2,3-bi- sphosphoglycerate-dependent phosphoglycerate mutase（BPGM）. Most of these proteins wich can be developed as the suitable vaccine candidate antigens need further research.