Expression of cauliflower mosaic virus gene I using a baculovirus vector based upon the p10 gene and a novel selection method.
A new baculovirus expression vector based upon the p10 gene of Autographa californica nuclear polyhedrosis virus (AcNPV) and a novel system for the screening of p10 recombinants have been developed. The insertion of a cassette containing the lacZ gene under the control of a heat-shock promoter of Drosophila melanogaster downstream from the cloning site in p10 transfer vectors allows the convenient identification of putative recombinants by virtue of their expression of beta-galactosidase. Using this p10 transfer vector an AcNPV recombinant was engineered with a cDNA copy of gene I of cauliflower mosaic virus (CaMV) in place of the p10 coding sequence. This p10 recombinant expressed CaMV gene I at levels equivalent to those of p10 and polyhedrin, and was shown to be as effective in producing this protein as recombinants exploiting the polyhedrin promoter. CaMV gene I protein formed large numbers of hollow fiber-like structures in the cytoplasm of infected cells. Because the polyhedrin gene remains intact, these p10 expression vectors may be exploited for the expression of heterologous proteins in insects infected per os and for the enhancement of baculovirus pathogenicity for insect control.
Vlak
J M
JM
Department of Virology, Agricultural University Wageningen, The Netherlands.
Schouten
A
A
Usmany
M
M
Belsham
G J
GJ
Klinge-Roode
E C
EC
Maule
A J
AJ
Van Lent
J W
JW
Zuidema
D
D
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D005814:Genes, Viral; D005822:Genetic Vectors; D007304:Insect Viruses / Q000235:genetics*; D007313:Insecta; D008969:Molecular Sequence Data; D009029:Mosaic Viruses / Q000235:genetics*; D015345:Oligonucleotide Probes; D010957:Plasmids; D015183:Restriction Mapping; D014162:Transfection
D004279:DNA, Viral; D015345:Oligonucleotide Probes