Coexpression of the B subunit of Shiga toxin 1 and EaeA from enterohemorrhagic Escherichia coli in Vibrio cholerae vaccine strains.

Infection and Immunity
J R ButtertonS B Calderwood

Abstract

A promoterless gene for the Shiga toxin 1 B subunit (stxB1) has been placed under transcriptional control of the Vibrio cholerae heat shock gene htpG. A chromosomal enterohemorrhagic Escherichia coli fragment containing eaeA and 400 bp of upstream DNA was added to the construct, downstream of stxB1; no transcription terminators were located between the two genes. The plasmid construct was confirmed by DNA sequencing; in vitro transcription-translation studies demonstrated expression of EaeA from the plasmid. The htpGp-->stxB1, eaeA construct was inserted into lacZ on the chromosome of Peru2, an El Tor V. cholerae strain with both attRS1 sequences and the entire cholera toxin genetic element deleted, and into lacZ in JRB10, a Peru2 derivative that has a second copy of htpGp-->stxB1 also inserted in the V. cholerae virulence gene irgA. Two plasmid constructs, one containing stxB1 under the control of the tac promoter and another containing htpGp-->stxB1,eaeA, were transformed into Peru2. Expression of StxB1 by these constructs was quantified by enzyme-linked immunosorbent assay and was highest in the plasmid construct with stxB1 under the control of the tac promoter. Localization of EaeA to the outer membrane of the vector strain...Continue Reading

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Citations

Apr 4, 2001·Journal of Travel Medicine·E T Ryan, S B Calderwood
Dec 14, 2002·Microbiology·Mark P StevensTimothy S Wallis
Aug 7, 2003·Expert Review of Vaccines·Sims K KochiUna S Ryan
Nov 4, 2004·Molecular Microbiology·Sarah M McLeod, Matthew K Waldor
Oct 25, 2000·Journal of Pediatric Gastroenterology and Nutrition·M M Levine
Oct 7, 2006·Current Opinion in Gastroenterology·E C Boedeker
Sep 23, 2006·Expert Review of Vaccines·Edward T RyanFirdausi Qadri
Nov 17, 2007·Biotechnology Letters·Anisia J SilvaJorge A Benitez
Jun 23, 2009·Journal of Bacteriology·Leyla Slamti, Matthew K Waldor
Jun 9, 2009·Journal of Bacteriology·William LainhartGerald B Koudelka
Dec 15, 2000·Journal of Bacteriology·J BehariS B Calderwood
Jul 17, 1998·Clinical Microbiology Reviews·J C Paton, A W Paton
Feb 11, 1998·Clinical Microbiology Reviews·J P Nataro, J B Kaper
Oct 21, 2004·Seminars in Pediatric Infectious Diseases·James P Nataro

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