Dissertation > Excellent graduate degree dissertation topics show

Cloning and Expression of the gB Gene of Porcine Cytomegalovirus of SiChuan Strain and the Analysis of Its Antigenicity

Author: ShiXiaoHong
Tutor: XuZhiWen；GuoWanZhu
School: Sichuan Agricultural University
Course: Preventive Veterinary Medicine
Keywords: Porcine cytomegalovirus gB gene Sequence analysis Cloning and expression Antigenic analysis
CLC: S852.659.2
Type: Master's thesis
Year: 2011
Downloads: 22
Quote: 0
Read: Download Dissertation

Abstract


Pig cytomegalovirus (PCMV) is a second pig herpes virus, were found to belong to the beta herpes virus subfamily, can cause pig inclusion body rhinitis disease. The disease since the first time in 1995 in the United Kingdom found distributed in pigs a wide range of serum antibody positive rate in Japan and Europe and the United States in more than 90%. Envelope protein gB as a necessary protein, plays an irreplaceable role in all herpes virus infection of the host cell adhesion, penetration, resulting in cell fusion and diffusion process, the protein has a good immune protection and immunogenicity. This experiment PCMV gB gene carried the following studies: 1. Pig cells Juhua virus gB's full clone and pET30-gB recombinant expression plasmid construct (1) primers were designed PCR amplification of the PCMV gB genes purpose fragments and original expression gene fragment. (2) the use of bioinformatics software analysis PCMV gB gene and the deduced protein function prediction, that the size of the PCMV gB gene was 2580bp, encoding 860aa, human herpes virus type 6 or 7 MCP gene closer relationship, the N-terminus of the peptide chain of 1-23 amino acids for the signal peptide, containing the transmembrane region and in between 729-751aa, preferably the overall hydrophilicity of the protein molecules, antigenic sites distributed in the entire sequence of the gB polypeptide chain. (3) EcoR Ⅰ and Sal Ⅰ restriction endonuclease digested the destination fragments and plasmid of pET30 () gB gene target fragment pET30 () to build fusion expression recombinant plasmid of pET30-gB. Found that when the separation and purification of recombinant plasmid pET30-gB expression in E. coli and the expression products IPTG-induced expression of the recombinant plasmid, recombinant plasmids in 37 ° C, IPTG concentration 0.6mmol / L induced 4h efficiently In addition, the experiment using His affinity chromatography separation and purification of the recombinant protein expression of the target protein, a molecular weight of about 80KD. Recombinant protein antigen 3.PCMV gB analysis of the purified protein rabbit polyclonal antibody, agarose diffusion experiments, the highest positive dilution titer of 1:16, showed that the the PCMV gB recombinant protein has a good immune immunogenicity. Western-Blot description recombinant PCMV gB protein has good reactogenicity. The completion of the study the PCMV gB gene cloning and more comprehensive bioinformatic analysis of the sequence and the deduced protein fusion gene PCMV gB, and preparation of the recombinant protein polyclonal antibody the antigenicity of the recombinant protein, which laid the foundation for further preparation of the high specificity and sensitivity PCMV genetic engineering antigen kit, also laid the foundation for the study of pig cytomegalovirus subunit vaccines.

Related Dissertations

  1. cDNA Cloning, Expression of vp5 and vp7 Genes and Subcecullar Localization of VP5 and VP7 Proteins in Grass Carp Reovirus,S941.41
  2. Cloning, Expression of vp6 and ns38 Genes and Immunogenicity of VP6 and NS38 in Grass Carp Reovirus,S941.41
  3. Survey in Nanjing and Analysis of the Mitochondrial Genome of Frankliniella Occidentalis (Pergande),S433
  4. Isolation and Identification of Low Pathogenicity Avian Influenza (H9) Virus in Henan and Study on the Biological Charactistics,S852.65
  5. Isolation and Characterization of Carbendazim-Degrading Strains, Cloning and Expression of the mheI Gene,X172
  6. Isolation and Identification of Infectious Bronchitis Virus and Sequence Analysis of Its S1 Gene and N Gene,S852.65
  7. Pathogenic Mechanism of Invasion Associated Genes in Avian Pathogenic E. Coli Duck Isolate DE205B,S852.61
  8. Isolation, Identification and Degrading Characteristic of a Cyhalofop-Butyl-Degrading Bacteria, and Cloning, Expression of a Cyhalofop-Butyl-Hydrolyzing Gene chbH,X172
  9. Cloning and Expression of Organic Solvent Tolerant Lipase Gene from Staphylococcus Saprophyticus M36,Q78
  10. Screening of Bacterial Strains with High α-Galactosidase Activity and Its Gene Cloning, Expression, Purification and Characterization,TQ925
  11. Cloning 1Dx5 Gene from the Xinjiang Wheat Variety and Constructing Its Expression Vector,S512.1
  12. Mitochondrial Genome Sequence Analysis of a Brassica Juncea Landrace,S565.4
  13. Effects of Diclazuril on G3PDH in Second-generation Merozoites of Eimeria Tenella,S858.31
  14. Molecular Biological Characteristics of Multiple Porcine Fc Gamma RⅡB Sub-isoforms Generated by Alternative Splicing,S828
  15. Identification of New Protective Antigens of Streptococcus Equi SSP. Zooepidemicus,S855.11
  16. Molecular Epidemiology about White Spot Syndrome in Jiangsu Province,S945.1
  17. The Cloing、Expression and Initial Assessment of Immunoprection of Thioredoxin Glutathione Reductase of Schistosomiasis Japonicum,R392
  18. The Proteomics Analysis of Response to Cadmium Stress in Radish (Raphanus Sativus L.),S631.1
  19. Molecular Cloning, Sequence nad Expression Analyses of Two Transcription Factors ZmC4HC3 and ZmNAC,S513
  20. Cloning and Expression Analysis of Cold-Related Genes of Ammopiptanthus Nanus,S793.9
  21. Invasive plant Eupatorium cold acclimation ICE1 gene cloning,S451

CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Basic Veterinary Science > Animal Microbiology ( Veterinary Microbiology, ) > Livestock Virology > Single - stranded DNA virus
© 2012 www.DissertationTopic.Net  Mobile