A hybrid baculovirus-bacteriophage T7 transient expression system.
A hybrid recombinant baculovirus-bacteriophage T7 expression system was developed for transient expression in insect cells of plasmids with foreign genes provided with a T7 promoter. The coding sequence for T7 RNA polymerase, with or without a nuclear localization signal, was inserted into the genome of Autographa californica nuclear polyhedrosis virus. Recombinant viruses stably expressed T7 RNA polymerase in insect cells. Upon transfection of infected insect cells with plasmids containing the genes for chloramphenicol acetyltransferase (CAT), the hepatitis B virus precore-, core- or e- antigens under control of the T7 promoter, transient expression of these genes was detected by ELISA. The results obtained indicate that this baculovirus/T7 system provides a simple and widely applicable tool for transient gene expression studies.
van Poelwijk
F
F
Department of Virology, Binnenhaven, The Netherlands.
Broer
R
R
Belsham
G J
GJ
Oudshoorn
P
P
Vlak
J M
JM
Goldbach
R W
RW
D000818:Animals; D017123:Bacteriophage T7 / Q000235:genetics*; D016367:Baculoviridae / Q000235:genetics*; D002467:Cell Nucleus / Q000201:enzymology; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D003593:Cytoplasm / Q000201:enzymology; D004274:DNA, Recombinant; D012321:DNA-Directed RNA Polymerases / Q000235:genetics*; D015870:Gene Expression; D006512:Hepatitis B Core Antigens / Q000235:genetics; D006513:Hepatitis B e Antigens / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic; D021382:Protein Sorting Signals / Q000235:genetics; D011993:Recombinant Fusion Proteins; D018411:Spodoptera / Q000378:metabolism / Q000648:ultrastructure; D014162:Transfection; D014764:Viral Proteins
D004274:DNA, Recombinant; D006512:Hepatitis B Core Antigens; D006513:Hepatitis B e Antigens; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; D015500:Chloramphenicol O-Acetyltransferase; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases