Tufts University

Other

Harvard College

Andrew Camilli

John J Mekalanos

Eric J Rubin

Brian J Akerley

Claudette L Gardel

Minmin Yen

Stefan Schild

David W Lazinski

Lynne Cairns

Eric Jorge Nelson

Compositions and methods are provided for preventing, ameliorating, or treating a disease caused by a species of bacterial genus, for example, cholera caused by, the compositions containing two or more strains of lytic bacteriophage that infect and killcells. The bacteriophage are virulent, which replicate intracellularly and lyse and kill the bacteria. Use of two or more strains in a single treatment, as a result of a rate of mutation of the bacteria to simultaneous resistance to all of the bacteriophage to be so low as to be negligible, reduces appearance of phage-resistant bacteria to statistical negligibility. Normal human microbial flora species were not affected. In alternative embodiments of the method and the composition, antibiotic agents or other treatment agents can be administered with a cocktail of the plurality of bacteriophage strains.

Methods and compositions for preventing infection by aspecies

According to some aspects of the invention, provided herein are methods of amplifying nucleic acids using homopolymer-mediated ligation. The methods, in some embodiments, comprise adding a first homopolymer of at least 12 nucleotides to each 3' end of blunt-ended double-stranded nucleic acid containing a target nucleic acid, thereby producing a partially double-stranded nucleic acid.

Homopolymer mediated nucleic acid amplification

The methods and compositions of the present invention are directed to a vaccine againstcomprised ofouter membrane vesicles (OMVs). Such vaccines are relatively stable, facilitating distribution. Inventive methods generally include administration of a vaccine againstby intranasal, intraperitoneal, oral or intragastric routes. Such vaccines confer immunity to the individual, and when administered to pregnant subjects, can be conferred to the offspring of individuals.

Cholera vaccines

The invention features a general system for the identification of essential genes in organisms. This system is applicable to the discovery of novel target genes for antimicrobial compounds, as well as to the discovery of genes that enhance cell growth or viability.

Systematic identification of essential genes by in vitro transposon mutagenesis

vaccine strains which have a soft agar penetration-defective phenotype and methods for making such strains are described. Also described are methods for identifying new genes involved in,motility and the cloning, identification, and sequencing of,motB and fliC genes.

Vibrio cholerae mutants which are soft-agar penetration defective and lack a functional CtxA subunit

A reporter system relating to in vivo expression technology was devised to aid in the identification and study of genes that display temporal or spatial patterns of expression during infection of host tissues. The method of this invention comprises constructing a strain or pool of strains of a microorganism which contains an artificial cointegrate comprising a reporter gene flanked by direct repeats of sequences to which a resolvase enzyme binds, thus catalyzing excision of the reporter gene, and further contains a coding sequence under the control of a promoter sequence which encodes transcripts, the expression of which are easily monitored in vitro and which result in a permanent genetic change, excision of the reporter gene, that is heritable and easily detectable subsequent to induction of the synthetic operon.

Method of detecting gene expression

A reporter system relating to in vivo expression technology was devised to aid in the identification and study of genes that display temporal or spatial patterns of expression during infection of host tissues. The method of this invention comprises integrating a site-specific DNA recombinase expression vector, and a reporter gene that is permanently removable by the recombinase, by way of homologous recombination into a microorganism's chromosome and inducing the expression of a synthetic operon which encodes transcripts, the expression of which are easily monitored in vitro and which result in a permanent genetic change, excision of the reporter gene, that is heritable and easily detectable subsequent to induction of the synthetic operon.

Growing community of inventors

Sharon, MA, United States of America

Andrew Camilli