Nucleotide sequences of moraxella catarrhalis genome

Abstract

The present invention provides the genomic sequences of a library of purified, polynucleotides, or their complements, comprising the genome of Moraxella catarrhalis. The invention also provides the identification of open reading frames contained within the polynucleotides of the library. The present invention further provides for the use of the polynucleotides, their complements or fragments, and proteins or portions thereof for identifying ligands and useful diagnostic and therapeutic compositions. In addition the invention provides for vectors, host cell sand methods for producing M. catarrhalis proteins or portions thereof.

Description

[0001] This application is a continuation of U.S. patent application Ser. No. 09 / 596,002, filed on Jun. 16, 2000, which claims priority under 35 U.S.C. .sctn.119(e) to U.S. Provisional Application Serial No. 60 / 140,121, filed Jun. 18, 1999, both of which are hereby expressly incorporated herein by reference in their entireties.[0002] A portion of the disclosure of this patent document contains material which is subject to copyright protection. The copyright owner has no objection to the facsimile reproduction by anyone of the patent document or the patent disclosure, as it appears in the Patent and Trademark Office patent file or records, but otherwise reserves all copyright rights whatsoever.[0003] The present invention discloses nucleotide sequences from the genome of Moraxella catarrhalis. These sequences may be used in various assays and in the development of diagnostic and therapeutic agents.Sequence Listing[0004] The present application is being filed along with duplicate copi...

AI Technical Summary

Benefits of technology

[0028] "Derivative" refers to the chemical modification of a nucleic acid or amino acid molecule. Chemical modifications can include replacement of hydrogen by an alkyl, acyl, or amino group or glycosylation, pegylation, or any similar process which retains or enhances biological activity, stability, or lifespan of the molecule.

[0057] A nucleic acid molecule encoding a M. catarrhalis protein may be cloned into a vector and used to express the protein or portions thereof in host cells. The nucleic acid sequence can be engineered by such methods as DNA shuffling (U.S. Pat. No. 5,830,721) and site-directed mutagenesis to create new restriction sites, alter glycosylation patterns, change codon preference to increase expression in a particular host, produce splice variants, extend half-life, and the like. The expression vector may contain transcriptional and translational control elements (promoters, enhancers, specific initiation signals, and polyadenylated sequence) from various sources which have been selected for their efficiency in a particular host. The vector, nucleic acid molecule, and regulatory elements are combined using in vitro recombinant DNA techniques, synthetic techniques, and / or in vivo genetic recombination techniques well known in the art and described in Sambrook (supra ch. 4, 8, 16 and 17).

[0067] Various hosts including goats, rabbits, rats, mice, humans, and others may be immunized by injection with M. catarrhalis protein or any portion thereof. Adjuvants such as Freund's, mineral gels, and surface active substances such as lysolecithin, pluronic polyols, polyanions, peptides, oil emulsions, keyhole limpet hemacyanin (KLH), and dinitrophenol may be used to increase immunological response. The oligopeptide, peptide, or portion of protein used to induce antibodies should consist of about five to fifteen amino acids which are identical to a portion of the natural protein. Oligonucleotides may be fused with proteins such as KLH in order to produce antibodies to the chimeric molecule.

[0069] Alternatively, techniques described for the production of single chain antibodies may be adapted, using methods known in the art, to produce epitope specific single chain antibodies. Antibody fragments which contain specific binding sites for epitopes of the M. catarrhalis protein may also be generated For example, such fragments include, but are not limited to, F(ab').sub.2 fragments produced by pepsin digestion of the antibody molecule and Fab fragments generated by reducing the disulfide bridges of the F(ab').sub.2 fragments. Alternatively, Fab expression libraries may be constructed to allow rapid and easy identification of monoclonal Fab fragments with the desired specificity (Huse et al. (1989) Science 246:1275-1281).

Problems solved by technology

When host-microbe balance becomes disturbed, infection may ensue.

Some organisms are highly virulent and cause clinical illness when they colonize most or all hosts.

Alternatively, when host defenses are compromised, normally symbiotic microbes can induce serious, or even life-threatening, infections.

Failure or absence of appropriate host defense allows the bacteria to replicate and produce an inflammatory response in the alveoli.

Ampicillin, which had been universally effective in treating M. catarrhalis pneumonia, can no longer be used.