Expression of the tobacco mosaic virus movement protein using a baculovirus expression vector.
A cDNA clone of the tobacco mosaic virus 30K movement protein (MP) gene was constructed and introduced into an Autographa californica nuclear polyhedrosis baculovirus expression vector. Infection of Spodoptera frugiperda cells with the vector resulted in the synthesis of low levels of MP, which was detected by anti-MP serum as two closely related species of Mr approximately 34K and a third species of 32K. The authenticity of the recombinant MP was confirmed by comparison of the protein, on the basis of migration during SDS-PAGE, with authentic MP from several sources. It appeared that the recombinant MP was not modified by N-linked glycosylation, but was phosphorylated. The recombinant MP was produced in both a phosphorylated and an unphosphorylated state and the former species was shown to comigrate with plant-expressed MP during SDS-PAGE.
Atkins
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D
John Innes Institute, AFRC Institute of Plant Science Research, Norwich, U.K.
Roberts
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K
Hull
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R
Prehaud
C
C
Bishop
D H
DH
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D015153:Blotting, Western; D036022:Capsid Proteins; D002478:Cells, Cultured; D003001:Cloning, Molecular; D005822:Genetic Vectors; D009036:Moths; D011233:Precipitin Tests; D014027:Tobacco Mosaic Virus / Q000235:genetics*; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics*
D036022:Capsid Proteins; C009496:H protein, Tobacco mosaic virus; D014764:Viral Proteins