Dissertation > Excellent graduate degree dissertation topics show

The Preparaion and Separation of the Gecko Protein Composition

Author: LiLei
Tutor: XieYuAn
School: Guangxi Medical University
Course: Oncology
Keywords: Gecko Protein Gel chromatographic separation Gecko protein MTS Apoptosis c-myc p53 bax bcl-2
CLC: R285.5
Type: Master's thesis
Year: 2011
Downloads: 39
Quote: 1
Read: Download Dissertation

Abstract


Objective Gecko tissue proteins were extracted and separated and purified into a different molecular weight of the protein component. Fresh Gecko tissue minced, homogenized, repeated freezing and thawing three times high-speed centrifuged and the supernatant by filtration, ultrafiltration, and freeze-drying method to obtain the Gecko protein crude extracts. Dissolving the crude extract, centrifuged, and the supernatant was further purified and isolated through a SephadexG-50 gel filtration chromatography, and collecting the the Gecko protein component of the different molecular weight based on the separation curve, with the UV spectrophotometer and by SDS-PAGE and other methods to analyze the group content and molecular weight of the protein. Three protein peaks results Gecko protein extracted crude extract after SephadexG-50 gel chromatography mainly the collected three peaks Gecko protein components were named as group A, group B, C group. The content of the group A 44.8mg, B-group content of 206.18mg C group content of 126.38mg. SDS-PAGE electrophoresis showed that the molecular weight of the group A as 20KD-50KD, the B-group MW mainly for 3 kD-20KD C group of a molecular weight of less than 3 kD. The conclusions extracted protein after SephadexG-50 gel chromatography separation and purification main three groups Gecko protein components, molecular weight for 20KD-50KD, 3KD-20KD and less than the 3KD, of which the highest content of B group. The purpose of in vitro HepG-2 cells of liver cancer and leukemia K562 cell growth inhibition observed three groups of the Gecko protein component, filter out the strongest role in the Gecko protein component. MTS assay different concentrations of the three groups Gecko protein components HepG-2 cells and leukemia K562 cell growth inhibition after 48h, the strongest inhibitory effect on cell growth screened a group of Gecko protein components, further MTS assay after 72h of HepG-2 cells and leukemia K562 cell growth inhibition, biological inverted phase contrast microscope and fluorescence microscope after 48h HepG-2 cells and K562 cell morphology of cells withered mortality. Results the three groups Gejie protein fractions have a different degree of inhibition of the growth of HepG-2 cells and K562 cells, the higher the concentration, the stronger the inhibition. Molecular weight has significantly 3KD-20KD Gecko protein component HepG-2 cells and K562 cells strongest inhibition, compared with the control group, the difference was statistically significant. HepG-2 cells and K562 cells showed morphological changes of cell apoptosis 3KD-20KD the Gecko protein component after 48h after 48h, the apoptosis rate was higher, the difference was significant statistically significant. Conclusion Gecko protein components of HepG-2 cells and K562 cells with varying degrees of growth inhibition, which 3KD-20KD the Gecko protein components strongest inhibition in a dose-dependent manner by inducing HepG- 2 cell necrosis and apoptosis of K562 cells play a cell growth inhibition. Purpose vitro studies 3KD-20KD the Gecko protein component acting on HepG-2 cells and leukemia K562 cells after 48h of c-myc, p53, bax and bcl-2 gene mRNA and protein expression. Method 3KD-20KD the Gecko protein component role 48h HepG-2 cells and K562 cells for the experimental group, Gecko protein HepG-2 cells and K562 cells as the control group. RT-PCR was used to detect the control group and the experimental group of bax, bcl-2, cmyc of p53 the four gene mRNA expression, filter out the mRNA expression of different genes, and further by immunohistochemistry assay 3KD-20KD the Gecko protein group points affect its protein expression. Results (1) RT-PCR detection found 48h after 3KD-20KD the Gecko protein component role HepG-2 cells increased mRNA expression of bax gene bax mRNA expression increased in K562 cells, bcl-2 mRNA expression decreased, with the control group, the difference was significant. HepG-2 cells and K562 cells, the experimental group c-myc and p53 gene mRNA expression compared with the control group changed little, the difference was not statistically significant. (2) The results of immunohistochemistry 48h after 3KD-20KD the Gecko protein component role HepG increased bax protein-2 cells, K562 cells, increased expression of bax protein, bcl-2 protein expression decreased, compared with the control group, The difference was statistically significant. Conclusion Gecko protein components HepG-2 cell apoptosis induced by the increase of bax mRNA and protein expression levels necrosis, through the increase of bax mRNA and protein expression, to reduce the bcl-2 mRNA and protein expression, change 3KD-20KD bax / bcl-2 ratio of K562 cell apoptosis induced necrosis.

Related Dissertations

  1. Research on Combinatorial Regulation of Multiple Transcription Factors,Q78
  2. Recovery Whey Soy Protein by Microfiltration and Development of Whey Powder,TQ936.2
  3. Research on Disease Related miRNA Mining Based on Protein-Protein Interaction Network,R341
  4. The Research and Implementation of Protein Classification Algorithm on the Basic of String Kernel,TP301.6
  5. Study of Oridonin on SGC-7901 Cell Proliferation by Inhibitting Cell Cycle Proteins,R285
  6. Expression of hBMP4 and hBMP7 in Chinese Hamster Ovary Cells,Q78
  7. The Study of Color Change and Control Technology for Oyster Protein Beverage,TS254.4
  8. The Value of the Heart-Type Fatty Acid Binding Protein in Early Diagnosis of Acute Myocardial Infarction,R542.22
  9. Serum A-FABP Levels and Its Expression in Atherosclerotic Plaques of Diabetes Mellitus,R587.1
  10. The Regulate and Controlof Slug on P53-MDM2 System,R730.2
  11. Impact on Drosophila Brain Developement Through the Overexpression and Loss of Function of Gsα,Q75
  12. Study on Deodorization Techniques of Oyster Protein Beverage,TS254.4
  13. The Study on Cryopreservation and Mechanism of Freezing Injury on the Spermatozoa of Coelomactra Antiquate,S968.3
  14. Preliminary Study of Phosphorylation Effect on UGT1A3 Metabolism Activity Towards Drugs,R346
  15. The Anti-tumor Effect of CADPE and Inducing Apoptosis in Human Gastric Cancer Cells,R735.2
  16. Cloth and β- elemene combined administration of anti-tumor effect and mechanism of celecoxib,R96
  17. Study on Preparation and Properties of Protein Isolates from Tilapia,TS254.4
  18. Expression and Purification of Influenza a Virus M2 Protein and Detection of Its Antigenicity,R392
  19. Protein phosphatase 2A Cα subunit knockout cardiac energy metabolism due to remodeling,Q78
  20. Broiler Responses to Reduced-protein Diets Suppleme Nted with Synthetic Amino Acids and Small Peptides,S831.5
  21. The Preparation of Hpv58e7 Protein by GST-tagged Prokaryotic Expression System,R392

CLC: > Medicine, health > Chinese Medicine > Of Pharmacy > Pharmacology > Chinese medicine Experimental Pharmacology
© 2012 www.DissertationTopic.Net  Mobile