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Screening and Identification of Glycoprotein B Epitopes and Eukaryotic Expression of Glycoprotein D Derived from Herpes Simplex Virus-2

Author: JiangDeYu
Tutor: YuSanKe
School: Northwest University of Science and Technology
Course: Preventive Veterinary Medicine
Keywords: Herpes simplex virus type II Glycoprotein B Epitope screening Glycoprotein D Eukaryotic expression
CLC: R346
Type: Master's thesis
Year: 2010
Downloads: 59
Quote: 0
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Abstract


Type II herpes simplex virus ( Herpes simplex virus , HSV - 2 ) infection can be caused by a common and ancient sexually transmitted diseases - genital herpes , gonorrhea and syphilis , the incidence of sexually transmitted diseases after . 11 glycoprotein of HSV-2 gD can induce relatively high antibody levels , GB can induce neutralizing antibodies and cytotoxic T cell responses can cause therefore preferred subunit vaccine . Glycoprotein B ( glycoprotein B , gB ) as the total outer membrane protein of herpes simplex virus two subtypes in maintaining viral morphology and viruses enter the shrinkage of the main cells plays a very important role . And animal inoculation GB can induce high levels of neutralizing antibodies and can cause the protective T cell immune response . Therefore, select the HSV - 2 GB complete genome biology software to B and HLA - A * 0201 - restricted T cell epitopes were predicted , and Table B-cell screening potential in mice induced a high level of antibodies and mediated Th2-type immune response IgG1 subtype . Simultaneously , and two B cell epitopes induced antibody specificity and virus ( infection) . In addition , the use of immunological methods to detect whether the screening of T -cell epitopes from the gB able to stimulate the surface of T2 cells stably generate the HLA-A * 0201 complex ; / surgery can stimulate IFN produce effective screening of T -cell epitope in vivo - gamma and CD8 T cells in vitro. The results show that the predicted T -cell epitopes can stimulate the T2 cells stably produce the HLA-A * 0201 complex , and can stimulate T cells to secrete IFN - gamma and CD8 T cell cytotoxicity . HSV-2 333 strain genome DNA as a template was amplified encoding HSV-2 glycoprotein D ( glycoprotein D , Gd ) gene , and the vector pFastBac I is connected , transforming E.coil DH5a competent cells , and correctly by PCR and sequencing . The of of pFastBac I - GD Block recombinant plasmid to DH10Bac recombinant shuttle plasmid transfected Sf9 insect cells , recombinant baculovirus . Sf9 cells infected with recombinant baculovirus expressing the recombinant protein to optimize the expression conditions by SDS - PAGE and Western blot identification, successful expression of HSV - 2 GD protein . Laid the foundation for the next step .

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