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Title: CHARACTERIZATION OF THE EXPRESSION OF THE MAREK'S DISEASE VIRUS MEQ GENE IN A RECOMBINANT BACULOVIRUS

Author
item CUI, XIAOPING - AVIAN DISEASE ONCOL LAB
item WEI, PING - AVIAN DISEASE ONCOL LAB
item Lee, Lucy
item HOLLAND, MARGO - MICHIGAN STATE UNIVERSITY
item Silva, Robert
item KUNG, H. - UC DAVIS CANCER CENTER

Submitted to: World Poultry Congress Proceedings
Publication Type: Proceedings
Publication Acceptance Date: 8/20/2000
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: MEQ (MDV EcoQ) gene, present only in serotype I MDV, is perhaps the only gene that has a significant open-reading-frame expressed in all MDV transformed tumor cells. As such it is a strong candidate for the putative oncogene of MDV. The structure of MEQ also suggests its role in growth and oncogenesis. Because of the lack of an in vitro system to study its function, we cloned the MEQ gene from pcDNA3-MEQ into baculovirus vector pBlue Bac4 followed by cotransfection with Bac-N-Blue linear DNA into inset Sf9 cells. The recombinant virus was identified by picking blue plagues and verified by polymerase chain reaction for the 1.1 kb insert. One of the highly expressed recombinant virus containing MEQ was characterized and used for the development of monoclonal antibodies specific to MEQ. We obtained one such specific antibody and used it to characterize the expression of MEQ in SF9 cells. Immunofluorescence staining of recombinant baculovirus revealed intense staining in the nucleus. This indicated that the MEQ could direct the localization of its protein to the nucleus and suggests the nuclear localization signal of MEQ is fully functional. This monoclonal antibody also reacted spefically with recombinant fowlpox virus containing MEQ and with the wild type fowlpox virus in immunofluorescence staining. Western blotting analysis of recombinant baculovirus expressing MEQ revealed a protein about 55-70kDa. The abundently expressed MEQ in the recombinant baculovirus should provide a source of MEQ for functional analysis of MEQ in vitro and in vivo.