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Proteomics of Echinococcus Granulosus and Cyst Wall

Author: DuJuan
Tutor: ZhaoWei;WangZuoNa
School: Ningxia Medical University
Course: Genetics
Keywords: Echinococcus granulosus Proteomics Two-dimensional electrophoresis MALDI-TOF-MS
CLC: R392
Type: Master's thesis
Year: 2011
Downloads: 42
Quote: 0
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Purpose 1. Looking for a dimensional electrophoresis profiles applicable to research proteomics experiments Echinococcus granulosus get ideal Echinococcus former head larva wall tissue. 2, the use of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) to obtain protein peptide mass fingerprint the map (PMF). Bioinformatics technology former head larva, the wall of the protein component ratio analysis and understanding of the of Echinococcus granulosus protein composition and species, and these proteins function division. In the end, the role of anti hydatid disease vaccine research, a method of early diagnosis and new drugs echinococcosis target protein screening to provide research data. The grinding method processing samples organizations, by the Bradford method for protein quantitation. The two-dimensional electrophoresis separation in accordance with the different size of each protein in the sample and isoelectric point and molecular weight, the protein sample. Then the gel imaging scanners collection and save the two-dimensional electrophoresis image, and data analysis of protein spots using PDQuest software. Finally, the gel on the automatic plastic cutting instrument, cutting the target protein selected by software, plastic cutting instrument automatically cut collection colloidal particles. Micelles subject to the tryptic treatment, so that the proteolytic cleavage of a plurality of peptides. Then through the MALDI-TOF detection, for each protein, peptide mass fingerprinting (PMF) data. Than the protein that the PMF data with database data match, and preliminary identification of the nature of the protein components. Results. Former head larva, organized by two-dimensional electrophoresis wall chart, respectively isolated proteins of 233 and 107 points. 2 protein spots by MALDI-TOF-MS detection, the former head larva and wall were 106 of protein points and 42 protein spots success peptide mass fingerprinting the map (PMF). A total of 23 spots in the former head larva spots to be preliminary identification include: ① cytoskeletal proteins: actin, tropomyosin, myosin, paramyosin dynein activator protein. ② metabolism enzymes: peroxide reductase transketolase alcohol enzyme, phospholipase A2, glycogen synthase. ③ signal transduction proteins: calcium-binding protein, tyrosine protein kinase, serine / threonine protein phosphatase. ④ material transport protein: cytochrome C, ADP / ATP transport protein, fatty acid binding protein; preliminary identification of a total of 14 protein spots in the wall spots, they are: ① signal transduction proteins: calcium-binding protein, tyrosine protein kinase, serine / threonine protein phosphatase. ② metabolism enzymes: peroxide reductase transketolase alcohol enzyme phospholipase A2. (3) material transport proteins: cytochrome P450 fatty acid binding protein, apolipoprotein AI, and albumin. Conclusion 1. Summed set applies to the study of the Echinococcus granulosus proteomics experimental program, to get a better quality of the former head larva dimensional electrophoresis profiles, wall, respectively, 233 and 107 protein spots were isolated. Gets the former head larva, a wall of 106 and 42 protein spots peptide mass fingerprinting and related data. Preliminary identification Echinococcus granulosus former head larva wall tissue protein spots, which, former head larva in a total of 23 protein spots, 14 protein spots in the wall. These proteins may be related to the Echinococcus granulosus movement, growth, development, metabolism and regulation.

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