1. Expression of functional full-length hSRC-1 in eukaryotic cells using modified vaccinia virus Ankara and baculovirus
- Author
-
Arnaud Poterszman, Judit Osz, Natacha Rochel, Marc Ruff, Karine Pradeau-Aubreton, Dino Moras, Nathalie Troffer-Charlier, Robert Drillien, and Isabelle Kolb-Cheynel
- Subjects
Genetic Vectors ,Biophysics ,Vaccinia virus ,Biology ,medicine.disease_cause ,Transfection ,Biochemistry ,Virus ,Cell Line ,chemistry.chemical_compound ,Nuclear Receptor Coactivator 1 ,Cricetinae ,Protein purification ,medicine ,Animals ,Humans ,Molecular Biology ,Escherichia coli ,Insect cell ,Biological activity ,Cell Biology ,Molecular biology ,Recombinant Proteins ,Cell biology ,Steroid Receptor Coactivator 1 ,chemistry ,Nuclear receptor ,Vaccinia ,Baculoviridae - Abstract
Purified protein expression level and quality are contingent upon specific host expression systems. This differential production is particularly observed for proteins of high molecular weight, hampering further structural studies. We developed an expression method aimed at producing proteins in Escherichia coli, insect, and mammalian systems. Our novel protocol was used to produce in large scale the full-length 160-kDa steroid receptor coactivator 1 (SRC-1), a coregulator of nuclear receptors. The results indicate that we can produce biologically active human SRC-1 in mammalian and insect cells in large scale.
- Published
- 2012