Large-scale preparation of Shiga toxin 2 in Escherichia coli for toxoid vaccine antigen production
Article first published online: 27 JAN 2013
© 2013 The Societies and Wiley Publishing Asia Pty Ltd
Microbiology and Immunology
Volume 57, Issue 1, pages 38–45, January 2013
How to Cite
Arimitsu, H., Sasaki, K., Shimizu, T., Tsukamoto, K., Shimizu, T. and Tsuji, T. (2013), Large-scale preparation of Shiga toxin 2 in Escherichia coli for toxoid vaccine antigen production. Microbiology and Immunology, 57: 38–45. doi: 10.1111/j.1348-0421.2012.12004.x
- Issue published online: 27 JAN 2013
- Article first published online: 27 JAN 2013
- Accepted manuscript online: 15 NOV 2012 08:41PM EST
- Manuscript Accepted: 6 NOV 2012
- Manuscript Revised: 24 OCT 2012
- Manuscript Received: 11 SEP 2012
Additional supporting information may be found in the online version of this article.
|mim12004-sm-0001-suppfig.jpg||30K||Figure S1. (a) The expression plasmid of 6 x His tag-free Stx2 (pBSK-Stx2) was prepared from pBSK-Stx2(His) by replacing the first histidine-coding codon of the 6 x His gene to a stop codon (*) by site-directed mutagenesis using a QuikChange II Site-directed Mutagenesis Kit and a primer set of Stx2(His-)-f; 5′-CTGAAGTGCAGTTTAATAATGACTAACACCACCACCACCACTG-3′ and Stx2(His-)-r; 5′-CAGTGGTGGTGGTGGTGTTAGTCATTATTAAACTGCACTTCAG-3′. (b) Preparation of the cell lysate and western blot analysis were conducted as described in Materials and Methods. Lane 1; EHEC-derived Stx2 (500 ng), lane 2; lysate of the transformant with pBSK-Stx2 (corresponding to 1.5 × 108 cells).|
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