| Objective Haematogenesis, immune and stem /progenitor cells of the liversystem are mainly from the liver tissue of the embryo. Fetal liver cells werecommonly used in the clinic treatment of various diseases. Up to now, it isnot clear about the molecular mechanism of the liver embryogeny and theFLC therapeutics. Liver experiences changes in morphology andbiochemistry depend on protein expression during these stages. To discussthe whole rule of change about protein in the process of development onthe level of protein molecule and to find out the molecule base of fetal liveron haematogenesis and immune, We analyzed the proteome from the liverin the different developmental stages of embryo by comparative proteomics.Our aim is to established the base of action in the clinic application of FLCand treatment of congenital disease of liver.Methods1. Found the NIH mouse pregnant animal model, livers were separated from embryo mouse E12d, E13d, E14d, E16d, E18d and adult mouse, then immediately stored in -80℃.2. The livers were separated in the first dimension by isoelectric focusing in linear pH 3~10 immobilized pH gradient (IPG). The separation in the second dimension was performed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Coomassie Blue Staining of two-dimensional gel electrophoresis gels wereanalysised by PDQuest Software.3. Target protein spots were excised from the gel and the digested with trypsin. The MALDI-TOF mass spectrometer was operated in the reflector mode. A mass list of peptides was obtained for the target protein digest. Then peptide mass fingerprint was submitted to an appropriate software (Mascot: Peptide Mass Fingerprint) for identify the protein (http://www.matrixscience.com/cgi/search_form.).Results1. 2-D PAGE profile show that these proteins in the liver were located mostly in the range of mass 14.4~75.4 kDa and isoelectric points (pI) 4~8. 280,466,790,805,850 and 925 protein spots with a molecular mass were catalogued from Coomassie Brilliant Blue-stained gels of the E12d,E13d,E14d,E16d,E18d fetal liver and adult liver respectively. When E18d gel acted as master,The number of matched protein spots E14d,E16d and adult gel was about 57%,61% ,69% respectively.2. Fourteen differentially expressed protein spots were analysed by MALDI -TOF-MS. Six protein spots of them were identified clearly by sequence database searching. They are Cyclin-dependent kinases regulatory subunit 1 (CKS-1),Fanconi anemia group A protein homolog (FACA protein),Tyrosine-protein kinase BLK (p55-BLK),General transcription factor 3C polypeptide 4 (TF3C-delta), TBC1 domain family member 13 , Macrophage receptor MARCO (Macrophage receptor with collagenous structure). CKS-1 and FACA protein were detected in gels of E18 fetal liver and adult mouse liver, p55-BLK,TF3C-delta were significantly higher expressed on gel of E14d fetal liver, TBC1 domain family member 13 was detected in gels of E14d, 16d, E18d fetal liver and adult mouse liver, Macrophage receptor MARCO was significantlyhigher expressed in gels of E14d fetal liver and adult mouse liver.Conclusion1. The liver go through an evident change in the expression of proteins during the embryo development,the patterns and amount of proteins are increasing. These changes are relevant with the development, structure differentiation and function perfection of fetal liver.2. CKS-1 belongs to the CKS family. It can bind to the catalytic subunit of the cyclin dependent kinases(CDK) and is essential for their biological function. CDK is an important serine/threonine protein kinase. The main function of CDK is to start up the DNA copy and place a premium on cell mitosis. CKS-1 may play a key role in proliferation, growth of the fetal liver cell by regulation CDK.3. FACA protein belongs to a multisubunit complex and mainly expressed in karyon, and at lower levels in cytoplasm. It is the DNA repairing protein that may operate in a post-replication repairing or a cell cycle checkpoint function. It may be related with DNA repair and the mainten...
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