Dissertation > Excellent graduate degree dissertation topics show

Proteomics Initial Screening and Functional Identification of Lipoic Dehydrogenase of Mycoplasma Bovis

Author: SunWenJing
Tutor: ZhangXiuYing
School: Northeast Agricultural University
Course: Basic Veterinary Science
Keywords: M.bovis Two-dimensional electrophoresis Western blot Cloning and expression
CLC: S858.23
Type: Master's thesis
Year: 2012
Downloads: 77
Quote: 0
Read: Download Dissertation


Mycoplasma bovis is an important pathogens of infected cattle,which can cause a variety of diseases.including bovine pneumonia, mastitis, arthritis,keratoconjunctivitis and so on.Mycoplasma bovis is difficult to be eliminated in cattle farms.The pathogen has become an important increasing factor to threat the cattle industry in China,since the first report of pneumonia caused by Mycoplasma bovis was reported in China.The most present study of Mycoplasma bovis focuse on pathogenic immune and diagnosis of nucleic acids,Howere,the studies on Mycoplasma bovine protein immunogenicity and immune protection features are deficient.In this study,the whole proteins of mycoplasma bovis can be separated by two-dimensional electrophoresis (2-DE).Combinning with Western blot, the immune-related proteins can be screened out. Now,the sequence of the whole genome of M.bovis Hubei isolation was finished and annotated.It is the base of the exactly information about mmune-related proteins that can be obtained.The lipoic dehydrogenase which is the mmune-related protein was figer out. Lipoicde hydrogenase (LPDH) is a flavin oxidoreductasea,It is widespread in a variety of organisms, and plays an important role in energy metabolism. At present,a variety of bio-lipoic dehydrogenase genes have been identified,but few of Mycoplasma bovis lipoic dehydrogeenase genes were reported.The recombinant protein of M.bovis LPDH(M.LPDH), had a molecular weight of63kDa, was obtained by applying prokaryotic expression system. And through the optimization of prokaryotic expression system, we obtain the soluble M.LPDH.Wester blotting、Dot blotting and ELISA assays were used for immunology validation. The result of Wester blotting is negative,while the result of Dot blotting is postive.The result of ELISApreliminarily showed that the recombinant protein can reacted with the M.bovis positive bovine antiserum made by natural infection and experimental infection.The results reveal that M.LPDH maybe an immuno-related protein and a prospect antigen for diagnosis of M. bovis.The purified M.LPDH was used to generate Balb/c mouse polyclonal antibodies. This analysis, using anti-M.LPDH antibodies, revealed a strong reactivity to a protein of approximately63kDa that is found in the whole, membrane and soluble cytosolic protein fractions, suggesting that MbM.LPDH is present in both membrane and soluble cytosolic proteins of Mb cells.Although how Mycoplasma bovis adhere to the surface of host cell surface and the pathogenic mechanism are still not very clear,most scholars agree that the adherence of mycoplasmas to the host cell initiates infection. Mycoplasma bovis cell surface may exist related to lipid-associated membrane protein (of LAMPs), which can cause diseases.The use of confocal laser (confocal a laser scanning microscopy) technology can provide a clear view that the recombinant protein adhere to the cell surface of respiratory epithelium of cattle (EBL).It reveals that M.LPDH maybe a adhesion-related protein.All tests in this study, we determined that Mycoplasma bovis surface does exist lipoic dehydrogenase and has immunogenicity; Wester blotting and Dot blotting assays were used for immunology validation. The result of Wester blotting is negative,while the result of Dot blotting is postive; The results of ELISA show that the enzyme can be used as a candidate antigen to establish the diagnosis method; The results of confocal laser technology can hypothesized that the enzyme indeed plays a certain role in the process of bovine Mycoplasma adherece to the host cells. These conclusions for the further study of Mycoplasma bovis adhesion mechanisms, virulence, and vaccine development have important meanning.

Related Dissertations

  1. cDNA Cloning, Expression of vp5 and vp7 Genes and Subcecullar Localization of VP5 and VP7 Proteins in Grass Carp Reovirus,S941.41
  2. Cloning, Expression of vp6 and ns38 Genes and Immunogenicity of VP6 and NS38 in Grass Carp Reovirus,S941.41
  3. Cellular and Proteomic Analyses of Endocytosis Mechanism Involved in Pollen-tube Growth in Nicotiana Tabacum,Q942
  4. Proteomic Analysis of Vibrio Alginolyticus in Response to Four Types of Antibiotics,S941
  5. Isolation and Characterization of Carbendazim-Degrading Strains, Cloning and Expression of the mheI Gene,X172
  6. Pathogenic Mechanism of Invasion Associated Genes in Avian Pathogenic E. Coli Duck Isolate DE205B,S852.61
  7. Isolation, Identification and Degrading Characteristic of a Cyhalofop-Butyl-Degrading Bacteria, and Cloning, Expression of a Cyhalofop-Butyl-Hydrolyzing Gene chbH,X172
  8. Cloning and Expression of Organic Solvent Tolerant Lipase Gene from Staphylococcus Saprophyticus M36,Q78
  9. Screening of Bacterial Strains with High α-Galactosidase Activity and Its Gene Cloning, Expression, Purification and Characterization,TQ925
  10. Salt-Tolerance Evaluation of Brassica Napus Germplasm in the Bud and Seeding and Establishment of the Two-Dimensional Electrophoresis System,S565.4
  11. Effects of Diclazuril on G3PDH in Second-generation Merozoites of Eimeria Tenella,S858.31
  12. Isollation and Identification of Haemophilus Parasuis and Its Proteomic Analysis under in Vitro Growth Conditions Mimicking the in Vivo Environment,S858.28
  13. Molecular Biological Characteristics of Multiple Porcine Fc Gamma RⅡB Sub-isoforms Generated by Alternative Splicing,S828
  14. Identification of New Protective Antigens of Streptococcus Equi SSP. Zooepidemicus,S855.11
  15. The Cloing、Expression and Initial Assessment of Immunoprection of Thioredoxin Glutathione Reductase of Schistosomiasis Japonicum,R392
  16. The Influence of Early Intervention of Vitamina Combined with Mycobacterium Bovis Bacillus Calmette-Guerin on Lung CD11C+ and Anti-OVA Special IgE Expression in Adult SD Mice Asthma Model,R562.25
  17. The Proteomics Analysis of Response to Cadmium Stress in Radish (Raphanus Sativus L.),S631.1
  18. Molecular Cloning, Sequence nad Expression Analyses of Two Transcription Factors ZmC4HC3 and ZmNAC,S513
  19. Cloning and Expression Analysis of Cold-Related Genes of Ammopiptanthus Nanus,S793.9
  20. The Regulation and Significance of NHE1 in Acidic Micro-environment of Gastric Carcinoma,R735.2
  21. Mycobacterium smegmatis MSMEG 6 402 gene function,R378

CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Livestock, poultry, wildlife diseases > Livestock > Cow
© 2012 www.DissertationTopic.Net  Mobile