副猪嗜血杆菌外膜蛋白P2的提取及鉴定

Abstract

Abstract: In order to simplify theseparation steps of a single protein in the outer membrane protein P2 (OmpP2) of Haemophilus parasuis (Hps) and reduce the equipment requirements of membrane protein separation,Hps standard type 3 (SW114), 5 (Nagasaki) strains were used as experimental materials in this experiment.First, lysozyme, nucleic acid were used to enzyme digest enzymatic hydrolysis the Hps for its complete outer membrane. And then, using buffer containing 2% triton-X-100 for further disintegrate. At last, used the appropriate concentration of pancreatic enzymes to digest the strong hydrophobic protein. Finally, a rapid extraction of the Hps high-purity OmpP2 in vitro was found. The result of sodium dodecyl sulfate-poly propylene amide gel electrophoresis method (SDS-PAGE) showed that it was very consistent with our expection: The two strains of Hps OmpP2 was about 43 and 38 ku, respectively.The protease off peptides matched with Hps OmpP2 perfectly in mass spectrum (MALDI-TOF-TOF). This experiment had established a rapid extraction method of Hps OmpP2. It reduced the equipmental requirement and simplified the separation step of a single protein OmpP2 from the membrane protein. Both type 3 (SW114) and 5 (Nagasaki) strains OmpP2 had been found hybridization chromogenic reaction with serum collected from sick pigs in Dot-blot. It simplified that OmpP2 was an immunological antigens. This study would be worthy of the function and immune protective research for Hps OmpP2 and offered a new reference for Hps immunological diagnosis.

Key words: Haemophilus parasuis; outer membrane protein P2; extraction; identification

CLC Number:

S852.61

LUO Yin-zhu, XU Cheng-gang, ZHOU Su-ming, FENG Sai-xiang, HE Xian-hui, FAN Hui-ying, LIAO Ming, XIN Chao-an. Extraction and Identification of Outer Membrane Protein P2 from Haemophilus parasuis[J]. , 2014, 41(9): 225-230.

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Extraction and Identification of Outer Membrane Protein P2 from Haemophilus parasuis

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