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PMID: 2497580 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

High level expression of nonfused foreign genes with Autographa californica nuclear polyhedrosis virus expression vectors.

Virology ·Vol. 170 ·No. 1 ·1989-05-00 ·Pages 31-9

Luckow VA, Summers MD

Abstract

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.

MeSH Terms
Animals Cells, Cultured Chloramphenicol O-Acetyltransferase/genetics Gene Expression Regulation Genetic Vectors Glucuronidase/genetics In Vitro Techniques Insect Viruses/genetics Insecta Occlusion Body Matrix Proteins Promoter Regions, Genetic RNA, Messenger/genetics Recombinant Proteins/genetics Viral Proteins/genetics Viral Structural Proteins beta-Galactosidase/genetics
Chemicals
Occlusion Body Matrix Proteins RNA, Messenger Recombinant Proteins Viral Proteins Viral Structural Proteins polyhedrin protein, Nucleopolyhedrovirus Chloramphenicol O-Acetyltransferase beta-Galactosidase Glucuronidase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Luckow V A
Department of Entomology, Texas A&M University, College Station.
Summers M D
Article Info
Journal
Virology
Abbr.
Virology
ISSN
0042-6822
Published
1989-05-00
Pages
31-9
Language
English
Region
United States
NLM ID
0110674
Subset
IM
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