Expression of the Kirsten ras viral and human proteins in Escherichia coli.
AUTOR(ES)
Nakano, E T
RESUMO
The expression vectors pINIII-A and pINIII (lpp p5) were used to construct plasmids which direct the synthesis in Escherichia coli of the Kirsten ras viral (v-Ki-ras) and human cellular (c-Ki-ras) oncogene products as fusion proteins containing 9 and 10 extra amino acids, respectively, at their N termini. Authenticity of the bacterially produced proteins was determined by immunoprecipitation and immunoblot analyses with ras-specific monoclonal antibodies. After induction with isopropyl-beta-D-thiogalactopyranoside, the viral protein represented approximately 20% of the total cellular protein. The majority of the protein was found in the postsonication low-speed centrifugation pellet. The synthesized viral protein was active in GTP binding, as judged by autophosphorylation and photoaffinity labeling assays.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=253950Documentos Relacionados
- Molecular cloning of the temperature-sensitive 371 Kirsten murine sarcoma virus and expression in Escherichia coli of the mutant and wild-type viral Kirsten ras p21 proteins.
- Purification and characterization of human H-ras proteins expressed in Escherichia coli.
- Photoaffinity labeling with GTP of viral p21 ras protein expressed in Escherichia coli.
- Expression of the human erythrocyte glucose transporter in Escherichia coli.
- Expression of the human fibroblast interferon gene in Escherichia coli.