Dissertation > Excellent graduate degree dissertation topics show

Screening for Proteins Interacted with PrP Using Yeast Two-Hybrid in Mouse Brain cDNA Library

Author: SongBoCui
Tutor: ChenZhiBao
School: Heilongjiang Bayi Agricultural University
Course: Basic Veterinary Science
Keywords: Prion protein Yeast two hybrid system Screening co-immunoprecipitation
CLC: Q78
Type: Master's thesis
Year: 2010
Downloads: 129
Quote: 1
Read: Download Dissertation

Abstract


Transmissible spongiform encephalopathy (TSE) is a human and animal’s central nervous system degenerative disease. The typical feature is the acumulation of PrPsc which is abound with theβ-fold and possesses partial protease resistance according to the histopathology. PrPsc is from the PrPc that is abound withα-Helix, which mechanism is still unknown. PrPc is a high conservative protein that anchors to the cell membrane surface by glycosyl-phosphatidyl inositol anchor (GPI anchor) most expressed in neuron.Some researchers presume that there may be some other proteins affect in the pathogenesy of TSE, so searching for the proteins interacted with PrPc is significant for the research of the transform mechanism, and also the physiology function of PrPc.There are several technologies can be used to study protein-protein interaction, Yeast two-hybrid system has became one of the most drastic methods in the study of protein-protein interaction.The CytoTrap system uses the yeast S. cerevisiae temperature-sensitive mutant strain cdc25H,The cdc25 mutation present in the cdc25H strain prevents growth at 37℃, but allows normal growth at the permissive temperature (25℃). The CytoTrap system is based on the ability of the human Sos protein (hSos), to complement the cdc25 defect and to activate the yeast Ras-signaling pathway. Expression of hSos and its subsequent localization to the plasma membrane allows the cdc25H yeast strain to grow at 37℃. The localization of hSos to the plasma membrane occurs through the interaction of two-hybrid proteins. All the interactions of protein-protein happen in cytoplasm, so it can overcome the limitations of the traditionary yeast two-hybrid system.The prion gene subcloned into the pSos plasmid. The expression of the fusion protein PrP23-231 was not toxic to cdc25H and does not have self-activation were detected through transformation of the bait plasmid. The Cyto Trap yeast two-hybrid system could be used to fish the interacting proteins with PrP23-231 domain of mouse prion;At last, 1 positive clone was picked up among 10 putative positive clones after verification. By the extracting the plasmid from the positive clones, endonucleases digestion analysis and sequencing, it showed that Ras protein-specific guanine nucleotide-releasing factor 1(Ras-grf1).In order to validate interaction between protein Ras-grf1 and PrP23-231, we amplified co-immunoprecipitation to further validate interaction. The discoveries of the interaction protein may have important significances for the research of the transform mechanism of PrPc to PrPsc and elucidating the physiology function function of PrP.

Related Dissertations

  1. Study on Phylogenetic Diversity and Bioactivity of Symbiotic Fungi of Coral,R284
  2. Establishment of the Molecular Identifying System of Curvualria and Application in Difficult Species,Q949.32
  3. Study on the Screening and Properties of Degradation Coking Waste Water Advantage Strains,X172
  4. Screening of High Tield Protease Producing Strains and Research on Enzyme Characterization,TQ925
  5. Studies on the Techniques of Multi-season Cultivation for Nanling Mushroom,S646.11
  6. Incentive Mechanism of IT outsourcing based on principal - agent theory,F426.6
  7. Screening and Studying Plant Growth-promoting Potential of Managanese-resistant Strains,X173
  8. Screening and Identification of Zearalenone-degrading Strain and Optimization of Zen-degrading Conditions,TS201.3
  9. Interaction between Lrrc10 and Srf, and Cell Function Study of Nulp1,R541
  10. Study on the Function of FTH1 in the Course of Brucella Ovis 019 Infecting RAW264.7 Macrophages,S852.61
  11. Roles of Human Heart Developmental Candidate Genes ZNF382 and CXXC5,Q987
  12. Screening of Denitrifying Phosphate Accumulating Organisms from Printing and Dyeing Wastewater and Characteristics of Nitrogen and Phosphorus Removal,X172
  13. Preliminary Screening of Bioactivities of Enzymolysis Product of Walnut Protein,TS201.25
  14. Study on the Brewing Technique and Healthy Compositions of Sea-buckthorn Vinegar,TS264.22
  15. Study on Processing Technology and Efficacy Evalution of 30% Cangersu Emulsifiable Concentrate,S482.2
  16. Tianmu resistant Bacillus soil productivity Isolation and phylogenetic analysis,S154.3
  17. Confirmation and Functional Exploration of the Interaction between Angiogenin and Fibulin Protein Family,R735.3
  18. Construction on Enriched Microsatellite Library and Genetic Diversity Research in Swimming Crab (Portunus Trituberculatus),S917.4
  19. Analysis of Newborn Hearing Screening in Neonatal Intensive Care Unit,R764
  20. Screening of Strains with High Yield of HDMF and Optimization of the Fermentation Conditions,TQ655
  21. EOR properties of bacterial fermentation compatibility,TE357.9

CLC: > Biological Sciences > Molecular Biology > Genetic engineering (genetic engineering)
© 2012 www.DissertationTopic.Net  Mobile