Prokaryotic Expression Of The Genes GB,gI And Pp38 Of Marek's Disease Virus And Preparation And Application Of Their Specific Monoclonal Antibodies

Prokaryotic expression of the genes gB, gIand pp38 of Marek抯 disease virus andpreparation and application of their specificmonoclonal antibodiesPhD Candidate: Lingxia HanDirected by Prof. :Zhizhong CuiProf. :Aijian QinYangzhou University, Yangzhou ,Jiang Su,China,225009Marek抯 disease(MD) is a common and important neoplastie disease of chickens.,the causative pathogen belongs to serotype I which virulence is increasing.Much effort has been made to differenciate the pathotype of an isolate strain .Since no significance mutations on genomic level has been found involved with this change, certain changes in expression of virus proteins may be responsible for that. To facilitate comparison of the protein band patterns of the major proteins of Marek抯 disease virus (MDV) in virally infected cells using Western-blotting, monoclonal antibodies which can distinguish different virulent Type I MDV strains are needed.The recombinant plasmids containing gB, gI and pp38 were constructed using vector pGEX-6p-l for fusion expression in E.coli. The expressed proteins were purified using preparative SDS-PAGE and used to immunize the mice. Hybridoma cell lines secreting monoclonal antibodies specific for the gB. 01 and pp38 proteins, respectively, were screened by using enzyme linked immunosorbent assay,(ELISA) and indirect immunofluorescence antibody( IFA)..A total of 17 hybridoma cell lines secreting the specific antibodies were obtained from 16 fusions. The Mab 7C8 recognized the gB proteins of serotypes I and III of4MDV and were able to detect three specific protein bands of 1 OOkD, 6OkD and 49kD in MDV infected-CEFs by Western-blotting. The Mabs 2H3, 2H7 and 6F10 were gI-specific and detected one protein band of 32 kD on Western blots. The Mats 9GA4, 1G9, 80D5 and 4AD2 were against to pp38. Interestingly, these Mabs recognized not only serotype I, but serotype 111 as well in WA. This was the first report that serotype I and serotype III herpes virus of turke(HVT) shared common antigenic epitope(s).The above Mabs were valuable tools for studying MDV strains with diffrent vxrulences.