Overproduction of the protein encoded by the maize transposable element Ac in insect cells by a baculovirus vector

Charlotte Hauser, Heidi Fusswinkel, Jian Li, Cornelia Oellig, Reinhard Kunze, Markus Müller-Neumann, Manfred Heinlein, Peter Starlinger, Walter Doerfler*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

16 Scopus citations

Abstract

The polypeptide encoded in the Activator (Ac) element of Zea mays L. has been expressed in Spodoptera frugiperda insect cells using plasmids which carry the strong polyhedrin promoter of the baculovirus Autographa californica nuclear polyhedrosis virus (AcNPV). Recombinant AcNPVs with the Ac-cDNA integrated and under the control of the viral polyhedrin promoter have been isolated and their genomes have been partly characterized as to the location of the foreign DNA insert. Upon infection of S. frugiperda cells with the recombinant AcNPV, maize Ac element specific messenger RNAs, as well as a newly synthesized polypeptide with an apparent molecular weight of about 116 kDa, have been detected in extracts of recombinant infected cells. This polypeptide is absent from extracts of wild-type infected cells expressing the polyhedrin polypeptide which can be recognized by the presence of nuclear inclusion bodies. Recombinant infected cells lack this protein. The Ac specific polypeptide is detected by antisera, which have been raised against fusion proteins containing Ac sequences synthesized in Escherichia coli, both in immunoprecipitation and in Western blotting experiments. The Ac specific protein is a nuclear phosphoprotein and represents about 1%-2% of the newly synthesized protein.

Original languageEnglish (US)
Pages (from-to)373-378
Number of pages6
JournalMgg Molecular & General Genetics
Volume214
Issue number3
DOIs
StatePublished - Nov 1 1988

Keywords

  • Autographa californica nuclear polyhedrosis virus
  • Eukaryotic expression vector
  • Immunofluorescence
  • Plant gene expression in insect cells
  • Western blotting

ASJC Scopus subject areas

  • Genetics

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