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Nucleic Acids Research, 1989, Vol. 17, No. 4 1427-1443
© 1989

MOLECULAR BIOLOGY

Expression of biologically active middle T antigen of polyoma virus from recombinant baculoviruses

Jitka Forstová+, Nina Krauzewicz and Beverly E. Griffin

Department of Virology, Royal Postgraduate Medical School Du Cane Road, London W12 ONN, UK

Received December 8, 1988. Accepted January 26, 1989.

Two different recombinant baculoviruses have been generated for expressing the middle T antigen (MT) of polyoma virus in insect (Sf9) cells. One (pAcI-PyMT) produces moderate levels of MT and the other (pVL-PyMT) high levels. Indirect immunofluorescence and cellular fractionation studies with pAcI-PyMT infected Sf9 cells give results similar to those observed with wild type polyoma virus infected mouse cells, and show MT to be mainly associated with cytoplasmic membranes in the insect cell. In the latter, a sub-population of MT is phosphorylated in in vitro protein kinase assays. The yields of MT from pVL-PyMT infected cells are high enough to suggest that this protein can now be produced by this method in sufficient amounts for definitive biochemical and crystallographic analyses.

+Present address: Institute of Molecular Genetics, Czechoslovak Academy of Sciences, Flemingovo M.2, 16637 Praha 6, Czechoslovakia


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