Dissertation > Excellent graduate degree dissertation topics show

Screening of the Genes Highly Expressed in Tumors by mRNA Fluorescent Differencial Display

Author: WenYan
Tutor: WangWenBing
School: Jiangsu University
Course: Biochemistry and Molecular Biology
Keywords: malignant tumor fluorescence differential display fluorescence quantitative PCR Calcium binding protein C3orf1 gene baculovirus
CLC: Q78
Type: Master's thesis
Year: 2006
Downloads: 35
Quote: 0
Read: Download Dissertation

Abstract


Malignant tumor has at present become one of the most seriousdiseases that pose great theat to human’s health.Survey on death causesof Chinese population showed that malignant tumor is the most serviousdisease with one in five persons dying from malignant tumors. Commonmalignant tumors include; esophageal cancer, lung cancer, gastric cancer,leukemia and so on, all of which have become the focus of researchers inthe world.In this thesis, in order to investigate genes that express specificallyin malignant tumors, we studied gene expressed highly in esophagealcancer and gastric cancer as well the lung cancer cell line 95D with highmetastatic potential by fluorescence differential display (FDD). Thefollowing are what we have achieved and concluded:Thirty cDNA fragments were excised by performing FDD-PCRreactions with those cancer tissues and their respective normal tissuesadjacent to the tumor in esophageal cancer and gastric cancer, andtwenty-one differential bands were obtained by carrying out the secondPCR amplification. These fragments were cloned and sequenced, andnine cDNA fragments were obtained. Six of them come from esophagealcancer tissue, three from gastric cancer tissue. Seven cDNA fragmentsthat expressed more differentially than others were confirmed byfluorescence quantitative PCR, and the result showed that: five of theseven were significantly higher than that in the corresponding normaltissue adjacent to the tumor. The other two had no significantly difference.The result showed that it has highest homology (88.7%) with the calciumbinding protein of house mouse. Fluorescence quantitative PCR was used to detect the relative expression amount of it in six kinds of tumor cells,and the result showed that expression level of it in esophageal carcinomacell line TE1 was significantly higher than that of it in the other fivetumor cells(p<0.05) . The results above indicate that carcinogenesis oftumors may be associated with the specific expression of certain genes.Seven cDNA fragments were excised by FDD-PCR reactions withfrom lung cancer cell line with high metastatic potential (95D) and thelung cancer line(LTEP-a-2) . These fragments were cloned and sequenced,and three cDNA fragments were obtained. All of them come from the cellline 95D. Two cDNA fragments that expressed more highly than otherswere confirmed by fluorescence quantitative PCR in the cell line 95D.One gene highly homologous with the gene C3orf1 attracted our attention.The experimente get its span. The ORF of the gene was 858 bp, coding285aa. Expression level of genes of interest C3orf1 in some differenttumor cells was investigated by employing fluorescence quantitative PCR(p<0.05) .Both prokaryotic expression plasmid pET-28a-C3orf1 and baculovirusexpression transfer plasmid ETB-C3orf1 were successfully constructed,and the stable expression of C3orf1 in prokaryotic and baculovirusexpression systems was proved by SDS-PAGE.

Related Dissertations

  1. Establishment of Inderect ELISA for Detection of Antibodies Angainst STLV-1 and Preparation of Hybridoma Cell of Monoclonal Antibodies,R373
  2. Expression of VP2 Gene of Infectious Bursal Disease Virus and Preparation of Monoclonal Antibodies Against the Recombinant VP2 Protein,S852.65
  3. Analysis of the Differentially Expressed Genes in Fingered Citron under Cold Stress,S666.9
  4. Effects of the Therapeutic Communication System Intervention on the Postoperative Gynecological Malignant Tumor Patients with Depression,R473.73
  5. Analysis Distribution of Malignant Tumors in Kunshan, 2000-2010,R73-31
  6. Detection Technology of MicroRNA-21 for Non-small Cell Lung Cancer,R734.2
  7. Effects of ω-3 Fish Oil Fatty Emulsion Administration on the Influence of Acute Stress Response and the Function of Gastrointestinal Tract Recovery T in the Patients with Gastrointestinal Tract Malignant Tumor Surgery,R735
  8. Effects of Lactobacilli on the Gene Expression of Antimicrobial Peptide AvBD9 in Chicken Intestinal Epithelial Cells,S831
  9. Chinese medicine decoction of the cellular immune function of patients with malignant tumors,R273
  10. Epidemic Tendency for Malignancies in Tanghai County from 1995 to 2006,R73-31
  11. Ascites of Unknown Origin in Female Patients: A Retrospective Study of 132 Cases,R442.5
  12. Preliminary Research on Biological Features of Recombinant EG95-(C3d)3 Protein,S852.5
  13. Detection of Infetious Flacherie Virus(Chinese Isolate) by Nested RT-PCR and Expression of the Viral RNA Polymerase,S884
  14. Co-expression and Identified Bioactivity of VP1 Gen of FMDV Type O and Goat IL-18 Gene in Sf9 Cells,S852.5
  15. Construction and Identification of Recombinant Adenovirus or Baculovirus Expressing Structure Protein of Human Respiratory Syncytial Virus,R373
  16. Risk Factors for Nosocomial Infectious Complications after Abdominal Surgery for Malignant Disease,R735
  17. The Clinical Research of Pancreatic Cancer Patients Treated with 125I Radioactive Seed Implantation,R735.9
  18. The Constraction and Clinical Application of the Microcolumn Gel Immunodetection System in the Serum Tumor Marker-CA125 of Ovarian Malignant Tumor,R737.31
  19. Expression and Variance Analysis of FMDV Combining Gene in Bombyx Mori Baculovirus Expression Vector System,S852.65
  20. The Study of Ultrasonic Imaging in the Diagnosis of Abdominal Tumor of Children,R735
  21. Clinical Retropective Analysis of Malignant Ovarian Germ Cell Tumors,R737.31

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