Synthesis in Escherichia coli of the reovirus nonstructural protein sigma NS.

AUTOR(ES)

Richardson, M A

RESUMO

The coding region of reovirus type 3 genomic segment S3, encoding the nonstructural protein sigma NS, was placed under the control of the bacteriophage lambda pL promoter in the Escherichia coli expression plasmid pRC23 (J.C. Lacal, E. Santos, V. Notario, M. Barbacid, S. Yamazaki, H.-F. Kung, C. Seamans, S. McAndrew, and R. Crowl, Proc. Natl. Acad. Sci. USA 81:5305-5309). Derepression of the pL promoter led to the synthesis of a protein of the same molecular weight as sigma NS produced in reovirus-infected L cells. The expressed protein was indistinguishable from authentic sigma NS by peptide mapping with Staphylococcus aureus V8 protease and by immunoblot analysis. Most importantly, the purified protein had nucleic acid-binding properties similar to that previously shown for sigma NS obtained from infected cells. Binding of single-stranded RNAs by recombinant sigma NS protein was inhibited by GTP.

Documentos Relacionados

• Nucleotide sequence of reovirus genome segment S3, encoding non-structural protein sigma NS.
• The reovirus nonstructural protein sigma1NS is recognized by murine cytotoxic T lymphocytes.
• Quantitative control of the stationary phase-specific sigma factor, sigma S, in Escherichia coli: involvement of the nucleoid protein H-NS.
• Mammalian Reovirus Nonstructural Protein μNS Forms Large Inclusions and Colocalizes with Reovirus Microtubule-Associated Protein μ2 in Transfected Cells
• Efficient expression of influenza virus NS1 nonstructural proteins in Escherichia coli.