Expression of cauliflower mosaic virus gene I using a baculovirus vector based upon the p10 gene and a novel selection method
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Expression of cauliflower mosaic virus gene I using a baculovirus vector based upon the p10 gene and a novel selection method. / Vlak, Just M.; Schouten, Alexander; Usmany, Magda; Belsham, Graham J.; Klinge-Roode, Els C.; Maule, Andrew J.; Van Lent, Jan W.M.; Zuidema, Douwe.
In: Virology, Vol. 179, No. 1, 11.1990, p. 312-320.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Expression of cauliflower mosaic virus gene I using a baculovirus vector based upon the p10 gene and a novel selection method
AU - Vlak, Just M.
AU - Schouten, Alexander
AU - Usmany, Magda
AU - Belsham, Graham J.
AU - Klinge-Roode, Els C.
AU - Maule, Andrew J.
AU - Van Lent, Jan W.M.
AU - Zuidema, Douwe
N1 - Funding Information: We appreciated the expert techntcal assrstance of Joop Groene-wegen In electron microscopy. We acknowledge Dr. 1. Roosien for valuable suggestions and Dr. R. W. Goldbach for his continuous interest. The research of D.Z. is made possrble by a fellowship of the Royal Netherlands Academy of Arts and Sciences. Thus research was supported part by the Bromolecular Action Programme (Contract No. BAP-01 1%NL (Wageningen) and Contract No. BAP-01 19-UK (Pirbright)) of the commission the European Communities.
PY - 1990/11
Y1 - 1990/11
N2 - A new baculovirus expression vector based upon the pl0 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 IacZ 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 β-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 pi 0 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.
AB - A new baculovirus expression vector based upon the pl0 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 IacZ 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 β-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 pi 0 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.
UR - http://www.scopus.com/inward/record.url?scp=0024997847&partnerID=8YFLogxK
U2 - 10.1016/0042-6822(90)90299-7
DO - 10.1016/0042-6822(90)90299-7
M3 - Journal article
C2 - 2219726
AN - SCOPUS:0024997847
VL - 179
SP - 312
EP - 320
JO - Virology
JF - Virology
SN - 0042-6822
IS - 1
ER -
ID: 381228683