4512血清型副猪嗜血杆菌外膜蛋白提取及分析  

Extraction And Analysis of The Outer Membrane Protein Haemophilus Parasuis of serotypes 4,5,12

作　　者：臧莹安[1] 李婉雁[1] 庞木生[1] 李淼[2,3] 宋帅[2,3] 李春玲[2,3] 

机构地区：[1]仲恺农业工程学院动物科学系,广东广州510225 [2]广东省农业科学院兽医研究所,广东广州510640 [3]广东省兽医公共卫生公共实验室,广东广州510640

出　　处：《中国兽医杂志》2013年第8期27-30,共4页Chinese Journal of Veterinary Medicine

基　　金：广东省农业科技重大专项(2009A020101006);广州市农业科技重大专项(2009A1-E041);广东省兽医公共卫生公共实验室开放基金项目(GSKJ090202)

摘　　要：采用超声波裂解技术,用膜裂解剂十二烷基肌氨酸提取副猪嗜血杆菌(Haemophilus parasuis,Hps)3种血清型(4、5、12)的外膜蛋白(OMP),用Bradford方法测定OMP浓度,将OMP进行SDS-PAGE凝胶电泳,以观察这3种血清型OMP的蛋白质条带分布。结果显示,当超声波条件为:功率为96W,超声破碎时间3s,间隔时间5s,循环作用30min时,提取的OMP进行凝胶电泳结果较好,提取的OMP的平均浓度分别为0.163mg/mL、0.163mg/mL、0.167mg/mL。对提取方法的条件摸索为OMP的深入研究及阐述OMP对Hps的致病机理提供重要参考依据。Lauryl sarcosine is used to extract the outer membrane protein （OMP） of three Haemophi- lus parasuis （Hps） serotypes （4,5,12） by using ultrasonic cracking technology. With the method of Brad- ford to determine the concentration of OMP, OMP was taken for SDS-PAGE gel electrophoresis to observe the distribution of OMP in 3 serotyes. The picture of SDS-PAGE gel electrophoresis shows that the OMP extraction yield better results when the ultrasonic power is 96 W, sonication 3 s, interval 5 s, keep cycling 30 min. The average OMP concentrations of 3 serotypes were 0. 163 mg/mL, 0. 163 mg/mL, 0. 167 mg/mL. This experiment has established a better extraction method of bacterial OMP and may provide an important reference for further research of the OMP, and its role in the pathogenesis of Hps.

关 键 词：副猪嗜血杆菌 外膜蛋白 SDS-PAGE 

分 类 号：S852.613[农业科学—基础兽医学]