Proteomics Research Of Low-Abundance Protein From Chicken Egg White During Chick Embryo Development

Study of Proteomics included the collection of Analytical Chemistry, Molecular Biology, Bioinformatics and other related frontier disciplines in the area of research. This work study on the changes of hen egg white proteome during specific biological processes by utilizes classical proteomics research tools. Comparative proteomic investigation of low-abundance proteins from hen egg white was explored during different incubations, using combinatorial peptide ligand libraries, two-dimensional electrophoresis, LTQ mass spectrometry, MALDI-TOF tandem mass spectrometry detection technology. To carry out this work was benefit in deep understanding of proteomic changes in low abundance proteins during chicken embryo development and biological function of low-abundance proteins from chicken egg white.The thesis was divided into eight chapters, the main contents of each chapters are summarized as follows:Establishment of a high throughput technique platform including combinatorial peptide ligand libraries technology with LTQ identification, found an applicability of the method in experimental basis for comparative proteomics investigations. Low-abundance proteins enrichment process optimized by examining the type of pH buffering system, different order of elution and egg white protein sample with different contents of buffer salt. Higher efficiency of combinatorial peptide ligand libraries in low-abundance proteins were obtained under free pretreated condition. LTQ linear quadrupole ion trap detection technology was performed in discrepancy SDS-PAGE bands identification. Combinatorial peptide ligand libraries technology with LTQ identification was suggested as a breakthrough currently used in limited basic research to better serve "foodomics" research. Feature detection in identifying low-abundance proteins from chicken egg white characterized as an evaluation of egg quality testing standards in applications.Applications of comparative proteomics in relative low-abundance proteins from hen egg white under chicken embryonic early development using combinatorial peptide ligand libraries technology, two-dimensional electrophoresis and matrix-assisted laser desorption-time of flight tandem mass spectrometry (Matrix-assisted laser desorption/ionization-time of flight, MALDI-TOF). For early hatching embryo has not yet formed a large number of features with the ability to absorb the egg white protein, egg white proteome changes relatively stable. It was found that relative abundance values of many protein precursor protein were significantly (p<0.01or<0.05) increase during early embryonic development phase. Meanwhile, it was found that some protein precursors were identified as new location coordinates on the two-dimensional electrophoresis maps. Thus, these biological function of low-abundance proteins were discussed and proposed.Exploration of low-abundance proteome from fertilized egg white on the last day of residue under chicken embryonic development. Expanding the two-dimensional electrophoresis of the observation area of low-abundance proteins comparing before and after combinatorial peptide ligand libraries treatment. In addition, comparative proteomic analysis of low-abundance egg white proteins during the rapid embryonic growth were curried out. It was found that the proteome of low-abundance proteins in incubation day16were more complexity in compare incubation day7.Relative abundance increase of proteins involved in five protein family, they were: Serpin Family; Clusterin Family; Transferrin Family and bacterial permeability-increasing protein (BPI) family; Lysozyme family. Relative abundance values were significantly decreased involved three protein families, they are:Serpin Family, Transferrin Family and bacterial permeability-increasing protein (BPI) family. According to such experiments, proposed and designed Chapter Ⅵ and Chapter Ⅶ of research ideas and content. It was found that these proteins may also play an important biological nervous system during embryonic development learning effect. Therefore, the proteome of low-abundance in hen egg residue egg white was different between early phase of embryonic development and the rapid growth period of embryonic development.Comparative proteomic analysis of low-abundance egg white proteins during the rapid embryonic growth period using peptide ligand library technology, two-dimensional electrophoresis, MALDI-TOF mass spectrometry techniques. The protein composition increased and the amount of albumin solids diminished in the residual egg white. There was a significant increase in the relative abundance of88protein spots (P<0.01and<0.05) over16days of incubation, representing11proteins from9protein families were identified. Moreover,16protein spots appear only in the egg white protein is completely absorbed after the last day of the two-dimensional electrophoresis spectra in controlled trials and not found in other groups. Of the three protein families:uPAR/Ly-6super family, histidine phosphatase family and OLFML3family5proteins. In addition, clusterin precursors were observed over a wide range of pH values and tenp protein increased continuously during embryonic development. Several of these dysregulated proteins were confirmed at the transcriptional level.Low-abundance proteins from two control groups were compared across different incubators in order to better understand the function of these proteins in egg whites. The relative increase in abundance of these differentially expressed proteins during the rapid embryonic growth period was different compared to store-incubated eggs, suggesting the pivotal role of hen egg white during embryonic development. These findings provide insights into regulating and supporting embryonic development during embryogenesis.Regularity analysis on low-abundance proteomic of chicken egg white during the whole process of embryo development based on PDQuest, NCBI data, GO and KEGG. Changes in the molecular function of low-abundance proteins in egg white is mainly divided into six species, while distributed in the cytoplasm13location area. Functional changes in egg white proteins covering:ion-binding protein, a lipid-binding protein, regulating the activity of the enzyme protein, hydrolyze activity, the key role in protein glycosylation, protein phosphatase activity, the enzyme-binding protein. These23process at least11egg whites biological development of low-abundance proteins involved in at least five low-abundance proteins involved in the transport process. Chick embryos detected the first16days of lysophosphatidic acid phosphatase type,,6metabolic pathways involved in riboflavin riboflavin metabolism-Gallus (chicken) fit≥75%. The study found that the combinatorial peptide ligand library technology advantaged in the exploration of low-abundance proteome changes in obvious. However, proteomic information may also missed due to limit of technology.Summarize multi-faceted research and explore multiple perspectives of variation of egg white proteome, low-abundance proteome through chicken embryonic development on the day0,3,7,10,14,16d hatching process. In addition, summary of shortage during experiments. Finally, prospects for further research and development of research strategy on chicken egg white, egg white protein and low-abundance proteome.