Dissertation > Excellent graduate degree dissertation topics show

Cloning and Eukaryotic Expression of MCM5 and Confirmation of MCM5 as a New P53 Interactor

Author: YangYaLin
Tutor: WuMian
School: University of Science and Technology of China
Course: Cell Biology
Keywords: p53 MCM5 Mass Spectrometry Protein - protein interactions GST pull-down Co-immunoprecipitation
CLC: Q78
Type: Master's thesis
Year: 2011
Downloads: 34
Quote: 0
Read: Download Dissertation

Abstract


P53 has been called the \P53 like the one responsible for processing and the integration of information and then orders the hub and control center, according to the type of cells facing stress stimulus and stimulus intensity differences, start the different downstream signaling pathways, and to determine the response of cells, that is, apoptosis, cell cycle arrest, cell senescence, DNA damage repair, cell metabolic state changes or autophagy. Different stress stimulus conditions may be the p53 after the occurrence of different forms of translation modification changes, thereby causing a series of cellular responses. P53 protein including the N-terminal (amino terminal) is a proline-rich transcription activation domain, DNA binding domain, a tetramerization domain and C-terminal (carboxy-terminal) domain. P53 response to a variety of stress conditions conducted post-translational modification occurs mainly in the N-terminus of the p53 protein molecules. Although the contents of some of the biological effects of the p53 transcription function, but the p53 transcription of their target gene activation is cell responses in a variety of stress stimulated a very important way. The amino terminus of the p53 protein not only with transcriptional activation, is also involved in the interaction between p53 and MDM2. Addition to MDM2 outside, there are a lot of interaction between p53 protein can affect p53 stability. In order to find new protein may regulate p53 transcriptional activation, we utilize include Human p53 protein amino-terminal 60 amino acids of the fusion protein the GST-p53N60 with HeLa cell lysates GST pull-down experiments by mass spectrometry to obtain a new p53-binding protein of MCM5 in. MCM5 replication as a DNA regulatory proteins, it is evolutionarily conserved eukaryotic cells minichromosome maintenance protein (minichromosome maintainance) one of the family members. This family protein (MCM2-7) may be formed an annular heterologous hexamer complex, and as a DNA helicase involved in DNA replication initiation and elongation process. HeLa cDNA library as a template for PCR, to obtain a full-length human MCM5 gene. And cloned into pEGFP-C1 and p3 * flag-myc-of CMV-24 two eukaryotic expression vector, and GST pull-down and immune coprecipitation experiments further confirmed that p53 and MCM2-7 helicase enzyme complex MCM5 interact, one of the regulatory subunit of the objects.

Related Dissertations

  1. Discovery and Biological Activities of Natural Michael Addition Acceptors,R284
  2. The Regulate and Controlof Slug on P53-MDM2 System,R730.2
  3. Metabonomics Study on the Usnic Acid and Erthromycin Estolate-induced Liver Toxicity by GC-MS,R965
  4. Study of Triptolide Induced Hepato-toxicology by MS-based Metabonomics Approaches,R285
  5. The Application of Eggshell Membrane for Solid-phase Extraction and the Determination of Trace Metal Elements in Environment Water,X832
  6. Study on Separation、 Preconcentration and Analysis of Trace Elements with Carbon Nanometer Materials,TB383.1
  7. Isollation and Identification of Haemophilus Parasuis and Its Proteomic Analysis under in Vitro Growth Conditions Mimicking the in Vivo Environment,S858.28
  8. Research of Screening and Function of P53 Regulators in KZNF (AxZ Type、SABZ Type and ACxZ Type) Family,R730.2
  9. Pharmacokinetics of Progesterone Cream in Beagle Dogs Determined with LC-MS/MS,R96
  10. Study on the Determination and Migration of Harmful Chemicals in Paper Food Packaging Materials,TS206.4
  11. Study on the Protein Composition of Intestinal Flora in the Antibiotics Interfered Mice and the Bacteria-related Diseases,R446.5
  12. Preliminary Research on the Interaction Between hELP3 and hELP4, yELP4, CTK1,Q75
  13. Study on the Matrix Solid-Phase Dispersion Applications of Pesticide Residues in Food,TS207.53
  14. Study on the Determination and Migration of Hazardous Chemicals in Polyethylene Plastic Food Packaging Materials,TS206.4
  15. Variation of Aroma Components in Jiupei of Fen Liquor during Fermentation,TS262.32
  16. Rapid Genotyping of ESBL Gene in Escherichia Coli and Drug Resistance Prediction,R446.5
  17. The Expression of PI3K and P53 in Prostate Cancer and Its Clinical Significance,R737.25
  18. Protein Expressions of EGFR、K-ras、P53 and Ki-67 in Colorectal Cancer, Gastric Cancer and Esophageal Cancer,R735
  19. Influence of NF-κB Inhibitor (PDTC) on Protein Expression of ASPP1 and ASPP2 of Non-small Cell Lung Cancer,R734.2
  20. Research of Expression and Clinical Significance of p53、PCNA and PTEN in Gastric Cancer,R735.2
  21. Quantiative Proteomics of Liver Immortalized Cell Line Chang Liver and Liver Cancer Cell Line HepG2,R735.7

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