Reconstitution of active recombinant Shiga toxin (Stx)1 from recombinant Stx1-A and Stx1-B subunits independently produced by E. coli clones
Toru Nishikawa
Shionogi Research Laboratories, Shionogi and Co. Ltd., Futaba-cho, Toyonaka-shi, Osaka 561-0825, Japan
Search for more papers by this authorJun Fujii
Department of Microbiology, School of Medicine, University of Occupational and Environmental Health, Yahatanishi-ku, Kitakyushu 807-8555, Japan
Search for more papers by this authorShin-ichi Yoshida
Department of Bacteriology, Faculty of Medicine, Kyushu University, Maidashi, Higashi-ku, Fukuoka 812-8582, Japan
Search for more papers by this authorCorresponding Author
Takashi Yutsudo
Shionogi Research Laboratories, Shionogi and Co. Ltd., Futaba-cho, Toyonaka-shi, Osaka 561-0825, Japan
*Corresponding author. Tel.: +81 (6) 6331-8081; Fax: +81 (6) 6331-8612, E-mail address: [email protected]Search for more papers by this authorToru Nishikawa
Shionogi Research Laboratories, Shionogi and Co. Ltd., Futaba-cho, Toyonaka-shi, Osaka 561-0825, Japan
Search for more papers by this authorJun Fujii
Department of Microbiology, School of Medicine, University of Occupational and Environmental Health, Yahatanishi-ku, Kitakyushu 807-8555, Japan
Search for more papers by this authorShin-ichi Yoshida
Department of Bacteriology, Faculty of Medicine, Kyushu University, Maidashi, Higashi-ku, Fukuoka 812-8582, Japan
Search for more papers by this authorCorresponding Author
Takashi Yutsudo
Shionogi Research Laboratories, Shionogi and Co. Ltd., Futaba-cho, Toyonaka-shi, Osaka 561-0825, Japan
*Corresponding author. Tel.: +81 (6) 6331-8081; Fax: +81 (6) 6331-8612, E-mail address: [email protected]Search for more papers by this authorAbstract
Escherichia coli clones expressing recombinant Shiga toxin (Stx)1-A and recombinant Stx1-B subunits, were established. Culture supernatants of these clones were examined for inhibitory activity on in vitro protein synthesis using luciferase as a reporter enzyme. Culture supernatant of the clone expressing Stx1-A, but not Stx1-B, showed the inhibitory activity. Neither recombinant Stx1-A nor Stx1-B showed Vero cell cytotoxicity. For reconstitution of biologically active toxin, the culture supernatants of the Stx1-A clone and the Stx1-B clone were mixed. The reconstituted recombinant Stx1 showed both Vero cell cytotoxicity and inhibition of in vitro protein synthesis.
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