High level expression of nonfused foreign genes with Autographa californica nuclear polyhedrosis virus expression vectors.
High levels of nonfused chloramphenicol acetyltransferase, beta-galactosidase, and beta-glucuronidase expressed under the control of new vector constructs of the polyhedrin promoter in Spodoptera frugiperda cells infected with Autographa californica nuclear polyhedrosis virus were investigated by SDS-PAGE and RNA dot blot analysis of total cytoplasmic RNA. When the polyhedrin ATG start codon was converted to ATT by site-directed mutagenesis, translation initiated at downstream ATG codons resulting in high yields of nonfused foreign proteins. When a stop codon was inserted downstream from and in phase with the polyhedrin ATG codon but upstream from the ATG of a foreign gene, nonfused proteins were also produced, but at lower levels. The level of steady-state polyhedrin gene-promoted mRNA was not affected by the mutation from ATG to ATT or the insertion of in phase stop codons downstream from the polyhedrin ATG.
Luckow
V A
VA
Department of Entomology, Texas A&M University, College Station.
Summers
M D
MD
D000818:Animals; D002478:Cells, Cultured; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D005966:Glucuronidase / Q000235:genetics; D066298:In Vitro Techniques; D007304:Insect Viruses / Q000235:genetics*; D007313:Insecta; D000077183:Occlusion Body Matrix Proteins; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics*; D014764:Viral Proteins / Q000235:genetics; D015678:Viral Structural Proteins; D001616:beta-Galactosidase / Q000235:genetics
D000077183:Occlusion Body Matrix Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins; C009829:polyhedrin protein, Nucleopolyhedrovirus; D015500:Chloramphenicol O-Acetyltransferase; D001616:beta-Galactosidase; D005966:Glucuronidase