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Brucellosis (Brucellosis, Brucellosis for short) is an animal-borne infectious disease caused by Brucella (Brucella) in our country is classified as a Class B infectious cause of human brucellosis and chronic infection of ruminants of infertility and miscarriage. Although brucellosis is an ancient infectious disease, but it is still very superficial understanding of, a lot of the problem has not been resolved, such as Brucella pathogenic mechanisms the parasitic mechanisms within the cell, immune mechanisms, different kinds of host parasitic difference, and brucellosis the immune prevention issues. With the development of bacterial proteomics research to analyze the distribution and function of the composition of the bacterial protein, to the disclosure of the above problems brought the gospel. Brucella membrane proteins having to promote adhesion to the surface, the transporter solutes and nutrition into the cell, output proteins and macromolecules, to allow the exchange of signals between cells, experience changes in the external environment, to maintain the stability of the film and relative to the outer periphery of the maintenance of intracellular The high osmotic pressure of the environment and other functions. Proteomics analysis Brucella membrane proteins, differences in comparing different kinds of membrane proteins, screening and identification of immunogenic protein, for the study of the biological classification of Brucella, the relationship between the different species, and explain the different kinds the host parasitic difference mining in Brucella virulence of immune-related molecules, to elucidate the pathogenesis of immune mechanisms, further research and development of new vaccines are important. This study with carbonates and new zwitterionic detergent method successfully extracted membrane protein of Brucella, separation of 2-DE protein separation effect. Virulent strain of Brucella species 544A and 16M comparative proteomics research found that the protein pI 4.0 to 7.0 MW 6.5 to 100kDa's, 2-DE gels were found differential expression of 130 points, liquid tandem mass spectrometry (LC-MS/MS) identified 31 protein spots, 23 ORF bioinformatics analysis, these proteins are involved in the conversion of energy generated, enzymes related to metabolism, signal transduction, cell membrane synthesis. The comparison shows that 544A and 16M with virulent Brucella bacteria, membrane protein metabolism in the host environment is significantly different: 1) both functional proteins generated by different energy conversion upregulated to escape to survive in the harsh environment of the host cell, the such as 544A gabD, gltA, BruAb2 0 325,16 M's BMEI0386 BME Ⅱ 0394; 2) amino acid metabolic pathways 544A ureC in a very the acid environment protection mechanism, 16M protective pathway the nitrogen deficiency environments BME Ⅰ 1635; 3) found in 544A methyltransferase family ubiG, and imaginary ABC transport cytoplasmic sugar-binding protein, Omp31 over the hydrogen peroxide environmental protective enzyme The the ahpC, general stress proteins BruAb1 1 942 proteins raised its environment to adapt to the intracellular survival survival conditions, which the ahpC, ureC for Brucella virulence factor known; 16M in up-regulated Dps has been proved 16M M5 and Rev1 Brucella strength of strain differentially expressed significantly possible for potential virulence factors. 4) the other two strains were found to have raised the hypothetical protein, such as 544A BruAb1 1 052 BruAb2 0 291,16 M BMEI1092, published differences the proteomics literature found different hypothetical protein, protein bioinformatics support its function can not be predicted, but suggests that Brucella large part of the hypothetical protein plays in its intracellular survival important role. Stronger strains of different pathogenic mechanism to conduct an in-depth study, the standard value for the understanding of the pathogenic mechanisms of Brucella. This study, while the 544A membrane protein with Brucella clinical positive cattle, sheep serum immune proteomics research. At pI of of 4.0 ~~ 7.0, MW 6.5 to 100kDa of 2-DE window, were found in the 19 common to the immunogenic protein, obtained by LC-MS/MS Mass Spectrometry and Mascot Search 19 ORF. 19 protein validation part of a known immunogenic protein, also newly found that the portion of the immunogenic protein, such as gltA, ubiG FABA, GAP, etc., wherein ubiG BruAb1 1 617 dadA 544A and 16M-oriented research differentially expressed proteins guess is that virulence factors can trigger the immune response. Preferably 5 protein gene for PCR primer design and amplified in 544A, 16M and respective vaccine strain S19, M5 5 immune-related protein gene amplification in both homology and immunogenicity, high interspecific suggesting that these proteins can be used for diagnosis and peptide vaccine research.
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