University System of Maryland

Swiss Serum and Vaccine Institute Berne

James B Kaper

Myron M Levine

Karen Jarvis

Bernadette Baudry-Maurelli

Alessio Fasano

Several EHEC proteins which are secreted into the culture supernatant have been discovered. These proteins are not produced by non-pathogenic , and produce a strong serum antibody response in patients with HUS and bloody diarrhea.

Escherichia coli secreted protein B

Several EHEC proteins which are secreted into the culture supernatant have been discovered. These proteins are not produced by non-pathogenic,, and produce a strong serum antibody response in patients with HUS and bloody diarrhea.

Immunodiagnostic test for enterohemorrhagic Escherichia coli infection

Vibrio cholerae 01 (CVD111) and non-01 (CVD112 and CVD112RM) serogroup

Method of isolating deletion mutants of Vibrio cholerae, wherein the deletion is predetermined by digestion with restriction endonucleases of known specificity. The deletions are inserted into the Vibrio cholerae chromosome by in vivo recombination between a plasmid carrying the desired deletion, with adjacent flanking sequences, and the Vibrio cholerae chromosome. The invention includes the isolation and characterization of a new Vibrio cholerae strain, CVD103Hg (ATTC No. 55,456), having a deletion in the tox gene, as defined by Acc I, Xba I, Cla I and/or restriction endonuclease sites, and carrying a mercury resistance gene. The invention also includes vaccines for protecting against the symptoms of cholera as well as methods for achieving this protection.

Vibrio cholerae strain CVD103Hg.sup.r, method of making same, and

Methods of isolating deletion mutants of Vibrio cholerae. In one method, the deletion is predetermined by digestion with restriction endonucleases of known specificity. The deletions are inserted into the Vibrio cholerae chromosome by in vivo recombination between a plasmid carrying the desired deletion, with adjacent flanking sequences, and the Vibrio cholerae chromosome. In another method, an initial in vivo recombination event of homologous sequences from the recombinant plasmid into the chromosome provides a selectable marker at this site. A second in vivo recombination event between homologous flanking sequences results in excision of proficient genes from the chromosome with the end product being a deletion mutation. Also provided are methods for the isolation and characterization of a new Vibrio cholerae strain having a deletion in the ctx gene, as defined by Acc I, Xba I, Cla I and/or restriction endonuclease sites and further having a deletion in the gene encoding zonula occludens toxin (zot).

Method of isolating restriction fragment deletions in Vibrio cholerae,

Method of isolating deletion mutants of Vibrio cholerae, wherein the deletion is predetermined by digestion with restriction endonucleases of known specificity. The deletions are inserted into the Vibrio cholerae chromosome by in vivo recombination between a plasmid carrying the desired deletion, with adjacent flanking sequences, and the Vibrio cholerae chromosome. The invention includes the isolation and characterization of a new Vibrio cholerae strain, (ATCC No. 55456), having a deletion in the tox gene, as defined by Acc I, Xba I, Cla I and/or restriction endonuclease sites, and carrying a mercury resistance gene. The invention also includes vaccines for protecting against the symptoms of cholera as well as methods for achieving this protection.

This invention relates to a method of isolating deletion mutants of Vibrio cholerae, wherein the deletion is predetermined by digestion with restriction endonucleases of known specificity. The deletions are inserted into the Vibrio cholerae chromosome by in vivo recombination between a plasmid carrying the desired deletion, with adjacent flanking sequences, and the Vibrio cholerae chromosome. The invention includes the isolation and characterization of a new Vibrio cholerae strain having a deletion in the tox gene, as defined by Acc I, Xba I, Cla I and/or Hind III restriction endonuclease sites.

Method of isolating restriction fragment deletions in vibrio cholerae,

This invention relates to a method of isolating deletion mutants of Vibrio cholerae, wherein the deletion is predetermined by digestion with restriction endonucleases of known specificity. The deletions are inserted into the Vibrio cholerae chromosome by in vivo recombination betweem a plasmid carrying the desired deletion, with adjacent flanking sequences, and the Vibrio cholerae chromosome. The invention includes the isolation and characterization of a new Vibrio cholerae strain having a deletion in the tox gene, as defined by Acc I restriction endonuclease sites.

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Columbia, MD, United States of America

James B Kaper