LKTA deletion mutant of P. haemolytica

Drug – bio-affecting and body treating compositions – Antigen – epitope – or other immunospecific immunoeffector – Bacterium or component thereof or substance produced by said...

Reexamination Certificate

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C424S234100, C435S173300, C435S252300, C435S069100

Reexamination Certificate

active

06331303

ABSTRACT:

TECHNICAL FIELD OF THE INVENTION
This invention is related to the field of bacterial genetics and more particularly to the field of respiratory pathogens of farm animals.
BACKGROUND OF THE INVENTION
P. haemolytica
as a pathogen causes serious economic damage to the animal farming industry. Vaccines which have been developed in an effort to control the disease have met with variable but limited success. Because the disease is caused in significant part by the animals' own reaction to
P. haemolytica
infection, inappropriately designed vaccines may actually worsen the clinical condition of infected vaccinates. Thus, there is a continuing need in the art for safe and effective vaccines which can reduce the morbidity and/or mortality of ruminants due to
P. haemolytica.
SUMMARY OF THE INVENTION
It is an object of the present invention to provide a
P. haemolytica
bacterium useful as a vaccine strain.
It is another object of the present invention to provide a method of inducing immunity to pneumonic pasteurellosis in ruminants.
It is an object of the present invention to provide a vaccine strain against pneumonic pasteurellosis.
Another object of the invention is to provide a ruminant feed.
Another object of the invention is to provide a temperature sensitive plasmid for manipulation of
P. haemolytica.
These and other objects of the invention are achieved by one or more of the embodiments described below. One embodiment of the invention provides a
P. haemolytica
bacterium which expresses no biologically active leukotoxin, expresses a form of leukotoxin molecule which induces antibodies which specifically bind to leukotoxin, and contains no foreign DNA.
Another embodiment of the invention provides a method of inducing immunity to pneumonic pasteurellosis in ruminants. A bacterium is administered to a ruminant. Immunity to the bacterium is thereby induced. The bacterium expresses no biologically active leukotoxin, expresses a form of leukotoxin molecule which induces antibodies which specifically bind to leukotoxin, and contains no foreign DNA.
Yet another embodiment of the invention provides a feed for ruminants. The feed comprises a bacterium which expresses no biologically active leukotoxin, expresses a form of leukotoxin molecule which induces antibodies which specifically bind to leukotoxin, and contains no foreign DNA.
Even another embodiment of the invention provides a vaccine for reducing morbidity in ruminants. The vaccine comprises a
P. haemolytica
bacterium which expresses no biologically active leukotoxin, expresses a form of leukotoxin molecule which induces antibodies which specifically bind to leukotoxin, and contains no foreign DNA.
Still another embodiment of the invention provides a temperature sensitive plasmid. The plasmid replicates at 30° C. but not at 40° C. in
P. haemolytica
. Moreover, it is of the same incompatibility group as the plasmid which has been deposited at the ATCC with Accession No. 98895.
The present invention thus provides the art with tools for genetically manipulating an agriculturally important pathogen. It also provides useful mutant strains which can be used effectively to reduce morbidity among ruminants, such as cattle, sheep, and goats, due to
Pasteurella haemolytica.


REFERENCES:
patent: 5422110 (1995-06-01), Potter et al.
patent: 5733780 (1998-03-01), Briggs et al.
patent: WO97 16531 (1997-05-01), None
patent: 97/41823 A (1997-11-01), None
George L. Murphy et al. “Hemolytic Activity of thePasteurella haemolyticaLeukotoxin” Infection and Immunity, Aug. 1995, pp. 3209-3212.
Natalie D. Fedorova & Sarah K. Highlander “Generation of Targeted Nonpolar Gene Insertions and Operon Fusions inPasteurella haemolyticaand Creation of a Strain that Produces and Secretes Inactive Leukotoxin” Infection and Immunity, Jul. 1997, pp. 2593-2598.
Fred M. Tatum et al. “Construction of an isogenic leukotoxin deletion mutant ofPasteurella haemolyticaserotype 1: characterization and virulence” Microbial Pathagenesis 1998; 24:37-46.
Robert E. Briggs et al. “Development and testing of a unique strain ofPasteurella haemolyticafor use in studies on colonization of the respiratory tract of cattle” AJVR, vol. 59, No. 4, Apr. 1998.
Robert E. Briggs et al. “Rapid spread of a unique strain ofPasteurella haemolyticaserotype 1 among transported calves” AJVR, vol. 59, No. 4, Apr. 1998.
Glynn H. Frank et al. “Colonization of the tonsils and nasopharynx of calves by a rifampicin-resistantPasteurella haemolyticaand its inhibition by vaccination” Am J Vet Res., vol. 56, No. 7, Jul. 1995.
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Beaumont et al. “Identification and Characterization of alcR, a Gene Encoding an AraC-Like Regulator of Alcaligin Siderophore Biosynthesis and Transport inBordella ppertussisandBordetella bronchiseptica” Journal of Bacteriology, Feb. 1998, pp. 862-870, vol. 180, No. 4.
Link et al. “Methods for Generating Precise Deltions and Insertions in the Genome of Wild-TypeEscherichia coli: Application to Open Reading Frame Characterization” Journal of Bacteriology, Oct. 1997, pp. 628-6237, vol. 179, No. 20.
Cotter and Miller “BvgAS-Mediated Signal Transduction: Analysis of Phase-Locked Regulatory Mutants ofBordetella bronchisepticain a Rabbit Model” Infection and Immunity Aug. 1994, pp. 3381-3390, vol. 62, No. 8.
Hamilton et al. “New Method for Generating Deletions and Gene Replacements inEscherichia coli” Journal of Bacteriology, Sep. 1989, pp. 4617-4622, vol. 171, No. 9.
Murphy G.L. et al. “Haemolytic activity of thePasteurella haemolyticaleukotoxin” Infection and Immunity, vol. 63, No. 8, Aug. 1995, pp. 3209-3212.
Cruz W.T. et al. “Deletion analysis resolves cell-binding and lytic domains of the Pasteurella leuktoxin” Molecular Microbiology, vol. 4, No. 11, Nov. 1990, pp. 1933-1939.
Petras S.F. et al. “Antigenic and virulence properties ofPasteurella haemolyticaleukotoxin mutants” Infection and Immunity, vol. 63, No. 5, Mar. 1995, pp. 1033-1039.

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