鸭疫里默氏杆菌ATP合成酶F1亚单位α亚基的部分生物学特性研究  

Study on some biological characters of ATP synthase F1 subcomplex alpha subunit of Riemerella anatipestifer

作　　者：邢林林[1,2] 卢凤英[2] 愈慧[2] 祁晶晶[2] 姜盼[2] 孙冰清[2] 崔俊生[2] 欧长灿[2] 于圣青[2] 常维山[1] 胡青海[2] 

机构地区：[1]山东农业大学动物科技学院,山东泰安271018 [2]中国农业科学院上海兽医研究所,上海200241

出　　处：《中国预防兽医学报》2015年第4期266-269,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金(31072156;31272590;31472224)

摘　　要：为筛选和鉴定鸭疫里默氏杆菌(RA)的感染相关蛋白,本研究采用血清1型RA CH3株活菌和灭活菌体免疫鸭的阳性血清分别与血清2型Th4株的全菌蛋白进行交叉免疫蛋白质组学分析,结果表明ATP合成酶F1亚单位α亚基(ATP-F1-α)、Gro EL蛋白和TPR repeat-containing protein等的抗体仅存在于活菌免疫鸭的血清中,推测其可能为感染相关的交叉性抗原;而Tbd R1的抗体在活菌和灭活菌免疫鸭的血清中均存在。利用自杀质粒同源重组的方法构建了ATP-F1-α的基因缺失株,并将其与野生株CH3的生长曲线进行比较。结果表明,缺失株菌体形态与野生株相似,但其在TSB培养基中的生长几乎停滞。以上结果表明RA ATP-F1-α既是一种交叉抗原,又与细菌的生长相关。To screen the potential infection-related protein of Riemerella anatipestifer, a two-dimensional electrophoresis （2-DE）-westem blot assay was performed to reveal the cross immunogenic antigens which occurred only in live bacteria- immunized duck sera, using live or inactivated R.anatipestifer serotype 1 strain CH3 immunized duck sera as the primary antibodies to detect total bacterial proteins of R.anatipestifer serotype 2 strain Th4. The results showed that antibodies against ATP synthase F1 subcomplex alpha subunit （ATP-F1-α, GroEL protein, and TPR repeat-containing protein, etc. occurred mainly in live bacteria-immunized duck sera, while antibodies against TonB-dependent receptor TbdR1 occurred in both live and inactivated R.anatipestifer immunized duck sera. Therefore, we deduced that the first three proteins were the potential infection-related antigens. Then, ATP-F1-α gene （atp-F1-a） of CH3 strain was deleted by a homologous recombination method, and the growth curves and morphologies of CH3△atp-F1-α mutant and the wild-type CH3 were compared. The results suggested that CH3△atp-F1-α mutant exhibited similar bacteria and colony morphologies as the wild-type CH3 did, and the growth of CH3△atp-F1-α mutant was almost stagnant in TSB medium. In conclusion, our results suggested that ATP-FI-a is a cross antigen of R.anatipestifer, and it is essential to the bacteria growth.

关 键 词：鸭疫里默氏杆菌 免疫蛋白质组学 ATP合成酶F1亚单位α亚基 交叉抗原 缺失 

分 类 号：S852.61[农业科学—基础兽医学]