PMID: 9634800Jun 1, 1995Paper

Improved adenovirus vector provides herpes simplex virus ribonucleotide reductase R1 and R2 subunits very efficiently

Bio/technology
B MassieY Langelier

Abstract

We have constructed a new adenovirus (Ad) expression vector, pAdBM5, that allows for the production of unprecedented levels of recombinant protein in the human 293 cell line using the Ad expression system. The main feature of this vector is a combination of enhancer sequences that increases the activity of the ectopic major late promoter (MLP) in recombinant Ad. In 293 cells infected with helper-free Ad recombinants generated with the pAdBM5 transfer vector, both herpes simplex virus (HSV) ribonucleotide reductase R1 and R2 subunits represent the most abundant polypeptides, accounting for as much as 15-20% of total cellular proteins. Our data suggest that this level of expression is probably very close to the upper limit of the system. Furthermore, when compared to the widely utilized baculovirus (Bac)/Sf9 expression system, the improved Ad vector showed a better performance for the production and purification of active HSV-2 ribonucleotide reductase R1 and R2 subunits. The R2 subunit was about 5-fold more abundant in recombinant Ad-infected 293 cells than in Bac-infected Sf9 cells while the R1 subunit was produced at roughly similar levels with either system. However, the amount of active soluble R1 obtained from recombinant A...Continue Reading

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Citations

Apr 1, 1999·Biotechnology and Bioengineering·J CôtéA Kamen
Apr 21, 1999·Current Opinion in Biotechnology·F Wurm, A Bernard
Nov 13, 2008·Cytotechnology·B MassieY Langelier
Oct 25, 2003·Nucleic Acids Research·Matt CottenKlaus Godl
Apr 16, 2003·Journal of Neurochemistry·Karolina WosikJosephine Nalbantoglu
Nov 5, 1999·Protein Expression and Purification·S C Makrides
Feb 10, 2007·Expert Review of Proteomics·Michael ForstnerLorenz M Mayr
Mar 3, 2004·Journal of Neuroscience Research·Sara C ShalinJ David Sweatt

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