High level expression of Streptococcus pyogenes erythrogenic toxin A (SPE A) in Escherichia coli and its rapid purification by HPLC.
The speA gene encoding streptococcal erythrogenic toxin A (SPE A) from Streptococcus pyogenes bacteriophage T12 was overexpressed in Escherichia coli under the control of the T7 promoter. Since most of the expressed protein was found in the periplasmic space, an osmotic shock extraction with 0.5 M sucrose resulted in a highly enriched preparation of SPE A. An additional two-step purification employing high pressure liquid chromatography resulted in a purified SPE A protein.
Yamamoto
M
M
Department of Microbiology and Immunology, University of Oklahoma Health Sciences Center, Oklahoma City 73190, USA.
Ferretti
J J
JJ
D001426:Bacterial Proteins; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005098:Exotoxins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification* / Q000494:pharmacology; D006801:Humans; D008213:Lymphocyte Activation; D008565:Membrane Proteins; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D017108:Streptococcus Phages / Q000235:genetics*; D013297:Streptococcus pyogenes / Q000821:virology*
D001426:Bacterial Proteins; D005098:Exotoxins; D008565:Membrane Proteins; D011994:Recombinant Proteins; C117980:SpeA protein, Streptococcus pyogenes; C027951:erythrogenic toxin