MXPA01002671A - Moraxella catarrhalis basb034 polypeptides and uses thereof - Google Patents

Abstract

The invention provides BASB034 polypeptides and polynucleotides encoding BASB034 polypeptides and methods for producing such polypeptides by recombinant techniques. Also provided are diagnostic, prophylactic and therapeutic uses.

Description

POL.PÉPT.DOS BASB034 OF MORAXELLA CATARRHALIS AND USES OF THEM FIELD OF THE INVENTION This invention relates to polynucleotides (hereinafter referred to as "BASB034" polynucleotide (s)), polypeptides encoded thereby (hereinafter referred to as "BASB034" or "BASB034 polypeptide (s)"), recombinant materials and methods for its production. In another aspect, the invention relates to methods for using said polypeptides and polynucleotides, including vaccines against bacterial infections. In a further aspect, the invention relates to diagnostic assays for detecting infection of certain pathogens.

BACKGROUND OF THE INVENTION Moraxella catarrhalis (also known as Branhamella catarrhalis) is a Gram negative bacterium frequently isolated from the human upper respiratory tract. It is responsible for several pathologies, the main ones being otitis media in babies and children and pneumonia in older people. It is also responsible for sinusitis, nosocomial infections and less frequently for invasive diseases. Otitis media is an important disease in children, both because of the number of cases and its potential sequelae. More than 3.5 million cases are registered each year in the United States, and it is estimated that 80% of children have experienced at least one episode of otitis before reaching the age of 3 years (Klein, JO (1994) Clin. Inf. Dis. 19: 823). If left untreated or chronic, this disease can lead to hearing loss that may be temporary (in the case of fluid accumulation in the middle ear) or permanent (if the auditory nerve is damaged). In babies, such hearing losses may be responsible for a slow learning to talk. Three bacterial species are mainly isolated from the middle ear of children with otitis media: Streptococcus pneumoniae, Haemophilus influenza (NTH¡) without type and M. Catarrhalis. They are present in 60 to 90% of cases. A review of recent studies shows that S. pneumoniae and NTHi both represent approximately 30% and M. catarrhalls approximately 15% of cases of otitis media (Murphy, TF (1996) Microbiol Rev. 60: 267). Other bacteria can be isolated from the middle ear. { H. influenza type B, S. pyogenes etc.) but at a much lower frequency (2% of cases or less). Epidemiological data indicate that, for pathogens found in the middle ear, colonization of the upper respiratory tract is an absolute prerequisite for the development of otitis; however, others have also required driving the disease (Dickinson, DP et al. (1988) J. Infect. Dis 158: 205, Faden, HL et al. (1991) Ann.Otorhinol.Laryngol.100: 612). These are important to activate the migration of bacteria to the middle ear through the eustachian tubes, followed by the initiation of an inflammatory process. These factors are unknown until now. It has been postulated that a transient anomaly of the immune system after a viral infection, for example, can cause the inability to control the colonization of the respiratory tract (Faden, HL et al. (1994Q J. Infecí Disa 169: 1312). Alternative explanation is that the exposure to environmental factors allows a more important colonization of some children, who subsequently are susceptible to the development of middle otiíis due to the permanent presence of paiogens in the middle ear (Murphy, TF (1996) Microbiol Rev. 60 : 267) The immune response of M. catarrhalis is poorly characterized The analysis of sequentially isolated strains of the nasopharynx of infants 0 to 2 years of age indicates that they frequently require and eliminate new strains.This indicates that an effective immune response against these bacferias it is set up by the colonized children (Faden, HL y oíros (1994) J. Infecí Disa 169: 1312) In most of the adults tested s, baclericidal antibodies have been idenified (Chapman, AJ and others, (1985) J. Infecí. Dis. 151: 878). Strains of M. catarrhalis showed variations in their ability to resist bacicide activity in the serum; in general, the isolates of sick individuals are more resistant than those who are simply colonized (Hol, C and oíros (1993) Lanceí 341: 1281, Jordán, KL et al. (1990) Am.J.Med. 88 (suppl. ): 28S). The resistance of serum, therefore, can be considered as a factor of virulence of the bacillary. An opsonization activity has been observed in the sera of children recovering from average hearing. The antigens identified by these different immune responses in humans have not yet been idenified with the exception of OMP B1, an 84 kDa proiein, whose expression is regulated by iron, and which is recognized by the sera of patients with pneumonia (Selhi, S, and oíros (1995) Infecí, Immun 63: 1516), and of UspA1 and UspA2 (Chen D. y oíros (1999), Infecí Immun 67: 1310). Some of the membrane proteins present on the M. catarrhalis surface have been characterized using biochemical methods, or for their potential involvement in the induction of proiebular immunity (for review, see Murphy, TF (1996) Microbiol, Reb.60: 267). . In a model of pneumonia of raion, the presence of antibodies developed with some of them (UspA, CopB) favors a more rapid elimination of the pulmonary infection. Olro polypepid (OMP CD) is alíamenie conserved eníre strains of M. catarrjhalis and presented homologies with a urine of Pseudomonas aeruginosa, which has been shown to be effective coníra es bacferia in animal models. The frequency of infections by Moraxella catarrhalis has risen dramatically in recent decades. This has been aided by the emergence of amphibian resis- phenefic strains and an increasing population of people with weak immune systems. It is no longer common to isolate Moraxella catarrhalis strains that are resistant to some or all of the standard antibiotics. This phenomenon has created a medical need that has not been met and a demand for new antimicrobial agents, vaccines, drug classification methods and diagnostic tests for this organism.

COMPENDIUM OF THE INVENTION The present invention relates to BASB034, in particular BASB034 polypeptides and BASB034 polynucleotides, recombinant materials and methods for their production. In another aspect, the invention relates to methods for using said polypeptides and polynucleotides, including the prevention and protection of microbial diseases, among others. In a further aspect, the invention relates to diagnostic assays for deifying diseases associated with microbial infections and conditions associated with other infections, such as assays for detecting the expression or activity of BASB034 polynucleotides or polypeptides.

Various changes and modifications in the spirit and scope of the present invention will be more readily apparent to those skilled in the art after reading the following descriptions and after reading the other parts of the present invention.

DESCRIPTION OF THE INVENTION The present invention relates to BASB034 polypeptides and polynucleotides as described in greater detail below. In particular, the invention relates to polypeptides and polynucleophides from BASB034 of Moraxella catarrhalis, which is related through amino acid sequence homology to Klebsiella pneumoniae, another outer membrane phospholipase A protein. The invention especially relates to BASB034 having the nucleotide and amino acid sequences disclosed in SEC.ID.NO:1, 3, 5 or 7 and SEC.ID.NO:2, 4, 6 or 8 respectively. It is understood that the sequences presented in the sequence listings below as "DNA" represent an illustration of one embodiment of the invention, as those skilled in the art will recognize that such sequences can be usefully employed in polynucleotides in general, including ribopolynucleotides.

Polypeptides In one aspect of the invention, Moraxella catarrhalis polypeptides, referred to as "BASB034" and "polypeptides," are provided.

BASB034"as well as its biological, diagnostic, prophylactic, clinical or ephemeriscally useful variants, and compositions comprising the same The present invention further provides for: (a) an isolated polypeptide comprising an amino acid sequence having at least 85% of identity, preferably at least 90% identity, preferably at least 95% denuded, most preferably at least 97-99% or exact identity, to that of SEQ ID NO: 2, 4, 6 or 8; (b) a polypeptide encoded by an isolated polynucleotide comprising a polynucleotide sequence having at least 85% identity, preferably at least 90% identity, preferably at least 95% identity, yet most preferably at least 97-99% or exact identity to SEQ ID NO: 1, 3, 5 or 7 over the length of SEQ ID NO: 1, 3, 5 or 7, respectively, or (c) a polypeptide encoded by an isolated polynucleotide that comprises of a polynucleotide sequence encoding a polypeptide having at least 85% identity, preferably at least 90% identity, preferably at least 95% identity, still most preferably at least 97-99% identity exact, to the amino acid sequence of SEC. ID. DO NOT. : 2, 4, 6 or 8. The BASB034 polypeptides provided in SEC. ID. NO .: 2, 4, 6 or 8, are the BASB034 polypeptides of Moraxella catarrhalis strains Mc2931 (ATCC 463617), Mc2908, Mc2913 and Mc2969. The invention also provides an immunogenic fragment of a BASB034 polypeptide, ie, a contiguous portion of the BASB034 polypeptide having the same immunogenic activity or substantially the same activity as the polypeptide comprising the amino acid sequence of SEQ. ID. NO .: 2, 4, 6 or 8.

That is, the fragment (if necessary when coupled to a carrier) is capable of raising an immune response that recognizes the BASB034 polypeptide. Said immunogenic fragment may include, for example, the BASB034 polypeptide lacking an N-terminal leader sequence, and / or a transmembrane domain and / or a C-terminal anchor domain. In a preferred aspect, the immunogenic fragment of BASB034 according to the invention, comprises sub-terminally the exfracellular domain of a polypeptide, which has at least 85% identity, preferably at least 90% identity, preferably at least less 95% identity, and most preferably at least 97-99% identity, so that SEC.ID.NO:2, 4, 6 or 8 over all the length of SEC.ID.NO:2. A fragment is a polypeptide that has an amino acid sequence that is complementary as well as does not cross the amino acid sequence of any polypeptide of the invention. As with the BASB034 polypeptides, the fragments can be "visible" or understood within a larger polypeptide from which they form a patch or region, most preferably as an individual conical region in a single larger polypeptide. Preferred fragments include, for example, endogenous polypepides which are a portion of an amino acid sequence of SEC.ID.NO:2, 4, 6 or 8 or their variants, as a coninuous series of residues including a sequence of amino-and / or carboxyl-terminal amino acid. Also preferred are degradation forms of the polypeptides of the invention produced by or in a host cell. Also preferred are fragments characterized by structural or functional alr bu buves, such as fragments comprising the alpha helix and alpha helix forming regions, beta sheet and beta sheet forming regions, spinning and spinning regions, regions of coil and coil formation, hydrophilic regions, hydrophobic regions, alpha-amphiphilic regions, bela-amphiphic regions, flexible regions, surface forming regions, subsymellation junction region and alionic regions. Other preferred fragments include an isolated polypeptide comprising an amino acid sequence having at least 15, 20, 30, 40, 50 or 100 contiguous amino acids from the amino acid sequence of SEC.ID.NO:2, 4, 6 or 8 , or an isolated polypeptide comprising an amino acid sequence having at least , 20, 30, 40, 50 or 100 contiguous amino acids truncated or deleted from the amino acid sequence of SEC.ID.NO:2, 4, 6 or 8.

Fragments of the polypeptides of the invention can be employed to produce the full-length polypeptide corresponding to peptide synsepsis; therefore, fragmenios can be used as inlermediates to produce the long-term polypeptides of the invention. Paríicularmenie preferred are varianíes where several amino acids 5-10. 1-5, 1-3, 1-2 or 1 are subsituted, eliminated, [or added in any combination. The polypeptides, or immunogenic fragments of the invention may be in the form of a "mature" prolein, or they may be derived from a larger protein such as a precursor or a fusion propion. It is usually desirable to include an additional amino acid sequence, which confers secretion or leader sequences, per-sequences, sequences that aid purification such as multiple histidine residues, or an additional sequence for stability during recombinant production. In addition, the addition of the exogenous polypeptide or lipid exome or polynucleotide sequences to elevate the immunogenic potential of the final molecule is also considered. In one aspect, the invention relates to genetically engineered soluble fusion proleins comprising a polypeptide of the present invention, or a fragment thereof, and several portions of conserved regions of heavy or light chains of immunoglobulins of various subclasses (IgG)., IgM, IgA, IgE). Preferred as an immunoglobulin is the consious partner of the heavy chain of human IgG, parficularly lgG1, wherein the fusion is présenla in the region of hinge. In a parficular embodiment, the Fc part can be moved simply through the incorporation of a cleavage sequence that can be separated with the coagulation factor Xa. In addition, this invention relates to processes for the preparation of these fusion proteins through genetic engineering, and their use to classify drugs, diagnosis and therapy. A further aspect of the invention also relates to polynucleotides that encode fusion proteins. Examples of fusion prolein technology can be found in International Patent Applications Nos. WO94 / 29458 and WO94 / 22914. The proteins can be chemically conjugated or expressed as recombinant fusion proteins allowing the production of high levels in an expression system, as compared to a non-fused protein. The fusion pattern can help provide T-helper epiploids (immunological fusion pads), preferably T-helper epiploids recognized by humans, or help to express the profine (expression enhancer) at higher levels than the native recombinant proiein. Preferably, the fusion pattern will be an immunological fusion partner as an expression enhancer pairing. The fusion periods include the D profiling of Haemophilus inlfuenzae and the non-structural prolein of the influenza virus, NS1 (haemagglutinin). Other fusion pattern is the protein known as LytA. Preferably, the C-terminal portion of the molecule is used. Lyta is derived from Streptococcus pneumoniae that synthesizes an N-acetyl-L-alanine amidase, amidase LytA (encoded by the lytA gene. {Gene, 43 (1986) pages 265-272.}.) An aufolysin that specifically degrades certain bonds in the base structure of the pyridoglycan. The C-lerminal domain of the LytA protein is responsible for the affinity to choline or some choline analogues such as DEAE. This property has been exploited for the development of plasmids expressing E. coli C-LytA useful for the expression of fusion proteins. The purification of hybrid proteins containing the fragment C-LylA at its amino terminus has been described. { Biotechnology: 10, (1992) p. 795-798} It is possible to use the repeat portion of the LytA molecule found at the C-ferminal terminus starting at residue 178, eg, residues 188-305. The present invention also includes variani of the above mentioned polypeptide annes, ie polypeptides which can vary from those present to conservative amino acid subslilutions, so one residue is suspended by another with similar characteristics. The lípicas of esía subsfituciones are enfre oirás Ala, Val, Ley and lie; between Ser and Thr; between the acid residues Asp and Glu; between Asn and Gln; and between the basic waste Lys and Arg; or aromatic residues Phe and Tyr. The polypeptides of the present invention can be prepared in any suitable form. Such polypeptides include isolated natural polypeptides, recombinantly produced polypeptides, synthetic polypeptides produced, or polypeptides produced through a combination of such methods. The means for preparing such polypeplides are well understood in the art. It is highly preferred that a polypeptide of the invention is derived from Moraxella catarrhalis, however, it can preferably be obtained from other organisms of the same taxonomic genus. A polypeptide of the invention can also be obtained, for example, from organisms of the same family or taxonomic order.

Polynucleotides It is an object of the invention to provide polynucleotides encoding BASB034 polypeptides, particularly polynucleotides that encode the polypeptide herein designated BASB034. In a particularly preferred embodiment of the invention, the polynucleotide comprises a region encoding BASB034 polypeptides comprising a sequence set forth in SEQ. ID. NO: 1, 3, 5 or 7, which includes a gene of full length, or a variant thereof. The BASB034 polynucleotides provided in SEQ ID NO: 1, 3, 5 or 7 are the BASB034 polynucleotides of Moraxella catarrhalis strains Mc2931 (ATCC 43617), Mc2908, Mx2913 and Mc2969. As a further aspect of the invention, isolated nucleic acid molecules encoding and / or expressing BASB034 and polynucleotide polypepides, particularly BASB034 polypepides from Moraxella catarrhalis and polynucleotides, including, for example, unprocessed RNAs, ribozyme RNAs, mRNAs, are provided. CDNAs, genomic DNAs, B and Z-DNAs. Other embodiments of the invention include biological, diagnostic, prophylactic, clinically or therapeutically useful polynucleotides and pplippeplins, and their variants, and compositions comprising them.

Another aspect of the invention relates to isolated polynucleotides, including at least one full-length gene encoding a BASB034 polypeptide having a deduced amino acid sequence of SEQ ID NO: 2, 4, 6 or 8 and polynucleotides closely related to the same and its variants. In another particularly preferred embodiment of the invention, BASB034 polypeptide of Moraxella catarrhalis comprising or consisting of an amino acid sequence of SEQ ID NO: 2, 4, 6 or 8, or a variant thereof is presented. Using the information provided herein, such as a polynucleotide sequence set forth in SEQ ID NO: 1, 3, 5 or 7, a polynucleotide of the invention encoding the BASB034 polypeptide can be obtained by utilizing standard cloning and classification methods, such as those to clone and sequence fragments of chromosomal ADM from bacleries using Caílin Moraxella catarrhalls cells as a parity maferial, followed by the obtention of a full-length clone. For example, to obtain a polynucleotide sequence of the invention, such as a polynucleolide sequence given in SEQ ID NO: 1, 3, 5 or 7, typically a collection of chromosomal DNA clones of Catlin Moraxella catarrhalis E. coli or some Another suitable host was rapidly chilled with a radiolabeled oligonucleotide, preferably a 17-mer or greater, derived from a partial sequence. Clones carrying DNA identical to that of the probe can then be distinguished using severe hybridization conditions. After sequencing the individual clones identified in this way by hybridization and designed sequencing primers of the original polypeptide or polynucleotide sequence of then, it is possible to extract the polynucleolide sequence in both directions to delermine a long-term gene sequence. Conveniently, such sequencing is carried out, for example, by using denatured double-stranded DNA, prepared from a plasmid clone. The appropriate techniques are described by Maniaíis, T., Frisích, E.F. and Sambrook and oíros, MOLECULAR CLONING, A LABORATORY MANUAL, 2a. Ed .: Cold Spring Harbor Laboraíory Press, Cold Spring Harbor, New York (1989). (see in parficular classification by Hybridization 1.90 and DNA templates of double-chain structure, denatured, and sequencing, 13.70). Sequencing of direct genomic DNA can also be performed to obtain a full-length gene sequence. Illustrative of this invention, each polynucleotide is fixed in SEQ ID NO: 1, 3, 5 or 7 and was discovered in a DNA library derived from Moraxella catarrhalis. In addition, each DNA sequence disclosed in SEQ ID NO: 1, 3, 5 or 7 contains an open reading frame that encodes a proiein having approximately the same amino acid residue as set forth in SEQ ID NO: 2, 4, 6 or 8 with a deduced molecular weight that can be calculated using the molecular weight values of amino acid residue well known to those skilled in the art.

The polynucleide of SEQ ID NO: 1, enriches the codon of paride at the nucleotide number 1 and the stop codon starting at the number of nucleotide 1327 of SEQ ID NO: 1, encodes the polypeptide of SEQ ID NO: 2. The polynucleotide of SEQ ID NO: 3, eny the start codon in nucleoid number 1 and the deink codon beginning at No. 1327 of SEQ ID NO: 3, encodes the polypeptide of SEQ ID NO: 4. The polynucleotide of SEQ ID NO: 5, enwrap the start codon at the number of nucleolide 1 and the stop codon starting at No. 1327 of SEQ ID NO: 5, encodes the polypeptide of SEQ ID NO: 6. The polynucleotide of SEQ ID NO: 7, between the start codon in nucleoid number 1 and the defense codon beginning at number 1327 of SEQ ID NO: 7, encodes the polypeptide of SEQ ID NO: 8. In a further aspect, the present invention provides an isolated polynucleotide comprising or consisting of: (a) a polynucleotide sequence having at least 85% identity, preferably at least 90% identity, preferably at least 95% identity; % identity, and most preferably at least 97-99% or identity exacerbates SEQ ID NO: 1, 3, 5 or 7 over all the length of SEQ ID NO: 1, 3, 5 or 7 respectively; or (b) a polynucleotide sequence encoding a polypeptide having at least 85% identity, preferably at least 90% identity, preferably at least 95% identity, and most preferably at least 97-99. % or 100% exaware the amino acid sequence of SEQ ID NO: 2, over all the length of SEQ ID NO: 2, 4, 6 or 8 respectively. A polynucleide encoding a polypeptide of the present invention, including homologs and orthologs from species other than Moraxella catarrhalis, can be obtained through a process comprising the steps of classifying an appropriate collection under severe hybridization conditions (e.g. temperature on the scale of 45-65 ° C and an SDS concentration of 0.1-1%) with a labeled or detectable probe consisting of or comprising the sequence of SEQ ID NO: 1, 3, 5 or 7 or a fragment of the same; and isolating a full-length genomic gene and / or clones containing said polynucleotide sequence. The invention provides an identical polynucleotide sequence over its entire length to a coding sequence (open reading frame) in SEQ ID NO: 1, 3, 5 or 7. Also, the present invention provides a coding sequence for a mature polypeptide or a fragment thereof, by itself as well as a coding sequence for a mature polypeptide or a fragment in a leukura frame with another coding sequence, as a sequence encoding a leader or secretion sequence, a sequence of pre, or pro, or prepro-prolein. The polynucleotide of the invention can also contain at least one non-coding sequence, including, but not limited to, at least one non-coding 5 'and 3' sequence, such as the transcribed sequences. but not induced, radiation signals (such as rho-dependent and rho-independence signals), ribosome binding sites, Kozak sequences, sequences that stabilize mRNA, introns, and polyadenylation signals. The polynucleolide sequence may also comprise an additional coding sequence encoding additional amino acids. For example, a marker sequence that facilitates the purification of the fused polypeptide can be encoded. In certain embodiments of the invention, the marker sequence is a hexa-histidine peptide, such as that provided in the pQE vector (Qiagen, Inc.) and described by Gentz et al., Proc. Natl. Acad. Sci., USA 86: 821-824 (1989), or an HA peptide tag (Wilson et al., Cell 37: 767 (1984), both may be useful for purifying the polypeptide sequence fused thereto. The invention also includes, but is not limited to, polynucleotides comprising a structural gene and its associated naphural sequences that encode expression of the gene.The nucleoid sequence encoding the BASB034 polypeptide of SEQ ID NO: 2, 4, 6 or 8 may be identical to the polypeptide coding sequence con? rmed in nucleosides 1 to 1326 of SEQ ID NO: 1, 3, 5 or 7, respectively.

Allernally, it may be a sequence, which as a result of the redundancy (degeneracy) of the genetic code, also encodes the polypeptide of SEQ ID NO: 2, 4, 6 or 8. The term "polynucleotide encoding a polypeptide" as used in the present invention encompasses polynucleotides which include a sequence encoding a polypeptide of the invention, particularly a bacterial polypeptide and more particularly a Moraxella catarrhalis BASB034 polypeptide by forming an amino acid sequence set forth in SEQ ID NO: 2, 4, 6, or 8. The term also encompasses polynucleotides that include an individual coninin region or discrete regions encoding the polypeptide (eg, polynucleotides interrupted by an intact phage, an infected insert sequence, an infected neighbor sequence, an incomplete transposon sequence, or June edition of RNA or genomic DNA reorganization) with additional regions, which It may also contain coding regions and / or which are not coding. The invention further relates to variants of the polynucleotides described herein that encode variants of a polypeptide having a deduced amino acid sequence of SEQ ID NO: 2, 4, 6 or 8. Fragments of polynucleotides of the invention can be used, for example , to synthesize polynucleotides from Iongifud complemen of the invention. Oftras particularly preferred embodiments are polynucleotides encoding variant BASB034, which has the amino acid sequence of the BASB034 polypeptide of SEQ ID NO: 2, 4, 6 or 8 where several, or some, or from 5 to 10, from 1 to 5 , from 1 to 3, 2, 1 or none of the amino acid residue are substiluted, modified, eliminated and / or added in any combination. Especially preferred, there will be subslilutions, additions, or silent eliminations, which will not ally the properties and activities of the BASB034 polypeptide. Ofras preferred embodiments of the invention are polynucleotides that are at least 85% identical in length to a polynucleotide encoding the BASB034 polypeptide having an amino acid sequence set forth in SEQ ID NO: 2, 4,6, or 8, and polynucleotides that are complementary to such polynucleotides. Alternately, highly preferred are polynucleotides which comprise a region that is at least 90% identical throughout its length to a polynucleotide encoding the BASB034 polypeptide and complementary polynucleolides thereto. In this respect, polynucleotides at least 95% identical to their length are particularly preferred. In addition, those with at least 97% are highly preferred among those with at least 95%, and among those those with at least 98% and at least 99% are particular and highly preferred, being highly preferred with at least 99% Preferred embodiments are polynucleotides that encode polypeptides that retain substantially the same biological function or activity as the mature polypeptide encoded by a DNA of SEQ ID NO: 1, 3, 5 or 7. According to certain preferred embodiments of this invention, there are provided polynucleophides which hybridize, particularly under severe conditions, to polynucleotide sequences BASB034 lales such as those polynucleolides in SEC.

ID NO: 1, 3, 5 or 7. The invention further relates to polynucleotides that hybridize to the polynucleotide sequences provided herein. In this regard, the invention especially relates to polynucleotides that hybridize under severe conditions to the polynucleotides described herein. As used herein, the terms "severe conditions" and "severe hybridization conditions" represent hybridization that occurs only if there is at least 95% and preferably at least 97% identity against the sequences. A specific example of severe hybridization conditions is incubation during the night at 42 ° C in a solution comprising: 50% formamide, 5x SSC (150mM NaCl, 15mM citrate friesic), 50mM sodium phosphate (pH7.6), 5x Denhardt's solution, 10% dextran sulfate, and 20 micrograms / ml salmon sperm DNA shared, denatured, followed by washing the hybridization soporie in 0.1x SSC at approximately 65 ° C. Hybridization and washing conditions are well known and will be Musilated in Sambrook, and other Molecular Cloning: A Laboratory Manual, 2a. Edition, Cold Spring Harbor, N.Y., (1989), particularly chapter 11 thereof. Hybridization of solution can also be used with the polynucleotide sequences provided by the invention. The invention also provides a polynucleide consisting of or comprising a polynucleotide sequence obtained by classifying an appropriate collection containing the complete gene for a polynucleotide sequence set forth in SEQ ID NO: 1, 3, 5 or 7, under conditions of severe hybridization with a probe that contains the sequence of said polynucleotide sequence disclosed in SEQ ID NO: 1, 3, 5 or 7, or a fragment thereof; and isolating such a polynucleotide sequence. Useful fragments for obtaining said polynucleotide include, for example, toylmene probes and initiators as described herein. As discussed herein with respect to polynucleotide assays of the invention, for example, the polynucleotides of the invention can be used as a hybridization probe for RNA, cDNA and genomic DNA to isolate full-length cDNAs and genomic clones that encode BASB034 and to isolate cDNAs and genomic clones from other genes that have an alias sequence identity, parficularly very alpha, for the BASB034 gene. Said probes will generally comprise at least 15 nucleotide residues or base pairs. Preferably, such probes will have at least 30 nucleotide residues or base pairs and can have at least 50 nucleotide residues or base pairs. Particularly preferred probes will have at least 20 nucleolide residues or base pairs and will be less than 30 nucleophilic residues or base pairs. A coding region of a BASB034 gene can be isolated through sorting, using a DNA sequence provided in SEQ ID NO: 1, 3, 5 or 7 to synthesize an oligonucleophil probe. A labeled oligonucleotide having a sequence complementary to that of a gene of the invention may then be used to classify a classification of cDNA, genomic DNA or mRNA to determine which member of the connection will hybridize the probe. Many methods are available and well known to those skilled in the art to obtain full-length DNAs, or extended-strain DNAs, for example, those based on the Rapid Amplification of cDNA End (RACE) method (see, for example, Forman, and others, PNSA USA 86) 5: 8998-9002, 1988). Recent modifications of the technique, illustrated by Marathon ™ technology (Clontech Laboratories Inc.) for example, have significantly simplified the search for longer cDNAs. In the Marathon ™ technology, the cDNAs have been prepared from mRNA extracted from a selected tissue and an "adapter" sequence linked to each effusion. Next, nucleic acid amplification (PCR) is performed to amplify the 5 '"missing" end of the DNA by using a combination of specific gene and specific adapter oligonucleotide primers. The PCR reaction is then screened by using "nested" primers, i.e., primers designed to heat and then cool within the amplified product (typically a specific initiator adapter that decays and further cools to 3 'in the adapter sequence and a specific primer). gene that heats and then chills an additional 5 'in the selected gene sequence). The products of this reaction can then be analyzed through DNA sequencing and long-length complemented DNA either by joining the product directly to the exisfenfe DNA to give a complete sequence, or by performing a full-length PCR separated using the new information from sequence for the 5 'primer design. The polynucleotides and polypeptides of the invention can be used, for example, as reagents and screening materials for the detection of diseases and diagnosis for diseases, particularly diseases of human beings, as described further in the present in relation to the tests of polynucleotide The polynucleotides of the invention which are oligonucleotides derived from a sequence of SEQ ID NO: 1-8 may be used in the processes of the present invention as described, but preferably for PCR, to determine whether the polynucleotides idenified in the present invention or all have been or not transcribed in bacteries in infected tissues. It is recognized that these sequences will also be useful in the diagnosis of the infection efapa and the type of infection that the pathogen obtains. The invention also provides polynucleotides that encode a polypeptide that is mature prolein plus additional amino- or carboxyl-terminal amino acids, or amino acids internal to the mature polypeptide (when the mature form has more than one polypeptide chain, for example). Such sequences can play an important role in the processing of a protein from the precursor to a mature form, they can allow the transport of proteins, they can enlarge or reduce the half-life of the protein or they can facilitate the manipulation of a protein for testing or production, among other things. As is generally the case in vivo, additional amino acids can be processed from the mature protein through cellular enzymes. For each and all polynucleotides of the invention, a polynucleotide complementary thereto is provided. It is preferred that these complementary polynucleotides are tofally complementary to each polynucleotide with which they are complementary. A precursor protein, which has a mature form of the polypeptide fused to one or more prosequences, can be an inactive form of the polypeptide. When the prosequences are removed said inactive precursors are generally activated. Some or all of the prosecutions can be removed after the affirmation. In general, the precursors are referred to as propropheins. In addition to the representations A, G, C, T / U, for nuclear nuclei, the term "N" can also be used to describe polynucleotide closes of the invention. "N" means that any of the 4 nucleotides of DNA or RNA may appear at said designated position in the DNA or RNA sequence, except if N is not a nucleic acid than when taken in combination with adjacent nucleotide positions, when read in the correction frame, it may have the effect of generating a premature termination codon in said lecfura frame. In summary, a polynucleotide of the invention can encode a mature protein, a mature protein plus a leader sequence (which can be termed a preprotein) a precursor of a mature protein having one or more prosequences that are not a leader sequence of a preprotein , or a preproprotein, which is a precursor to a proproprofein, comprising a leader sequence and one or more prosequences, which are generally removed during processing steps that produce acivive and mature forms of the polypeptide. According to an aspect of the invention, there is provided the use of a polynucleide of the invention for therapeutic or prophylactic purposes, in particular genetic immunization. The use of a polynucleotide of the invention in genetic immunization will preferably employ a suitable delivery method such as direct injection of plasmid DNA into the muscles (Wolff and others, Hum Mol Genet (1992) 1: 363, Manlhorpe and others, Hum. Ther. (1983) 4: 419), the supply of DNA in complex with specific proiein vehicles (Wu and Oros, J. Biol. Chem. (1989) 264: 16985), co-precipitating the DNA with calcium phosphate. (Benvenisty &; Reshef, PNAS USA, (1986) 83: 9551), encapsulation of DNA in various forms of liposomes (Kaneda et al., Science (1989) 243: 375), particle bombardment (Tang and Oros, Nature (1992) 356: 152 , Eisenbraun and others, DNA Cell Biol (1993) 12: 791) and in vivo infection using cloned relillar vectors (Seeger and others, PNSA USA (1984) 81: 5849).

Vectors, Host Cells, Expression Systems The invention also relates to vectors comprising a polynucleotide or polynucleotides of the invention, host cells that are genetically engineered with vectores of the invention and the production of polypeptides of the invention through techniques. recombinanies. It is also possible to employ cell-free irradiation systems to produce the profine proteins by using RNAs derived from the DNA constructs of the invention. The recombinant polypepides of the present invention can be prepared through processes well known to those skilled in the art from genetically engineered host cells comprising expression systems. Accordingly, in a further aspect, the present invention relates to expression systems comprising a polynucleotide or polynucleotides of the present invention, to host cells that are genetically engineered with said expression systems, and to the production of polypeptides of the invention. invention through recombinanid techniques. For recombinant production of the polypeptides of the invention, host cells can be genetically engineered to incorporate expression systems or portions thereof or polynucleotides of the invention. The inroduction of a polynucleotide into the host cell can be effected through methods described in many standard laboratory manuals, such as Davis et al., BASIC METHODS IN MOLECULAR BIOLOGY, (1986) and Sambrook et al., MOLECULAR CLONING: A LABORATORY MANUAL , 2a. Ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1989), such as, calcium phosphate transfection, DEAE-dextran-mediated transfection, transvection, microinjection, cationic lipid-mediated transfection, electroporiation, transduction, scraping charge, ballistic introduction and infection. Representative examples of appropriate hosts include bacterial cells, such as cells of syreptococci, siaphylococci, E. coli, syreptomyces, cuanobacteria, Bacillus subtilis, Neisseria meningitidis and Moraxella catarrhalis; fungal cells, such as yeast cells, Kluveromyces, Saccharomyces, a basidiomycete, Candida albicans and Aspergillus] insect cells such as Drosophila S2 and Spodoptera Sf9 cells; animal cells such as CHO, COS, HeLa, C127, BHK, 293, CV-1 and Bowes melanoma cells; and plant or vegetale cells, such as a gymnosperm or angiosperm cells. A wide variety of expression systems can be used to produce the polypeptides of the invention. Said vaccines include, among others, vectors derived from chromosome, episomes and viruses for example, vectors derived from bacterial plasmids, from bacteriophage, from transposons, from yeast episomes, from insertion elements, from chromosomal elements from yeast, from viruses such as baculoviruses, papovaviruses, viruses such as SV40, vaccinia virus, adenovirus, poultry positing virus, seudorabies virus, piconavirus, relrovirus, and alphaviruses and vectors derived from combinations thereof, such as those derived from gene elements of plasmid and bacleriophage , such as cosmids and phagemids. The expression system constructs may contain control regions that regulate as well as produce expression. In general, any system or vector suitable for maintaining, propagating or expressing polynucleotides and / or for expressing a polypeptide in a host can be used for expression to this respect. The appropriate DNA sequence can be inserted into the expression system through any of a variety of well-known techniques and ruin, such as, for example, those presented in Sambrook and others, MOLECULAR CLONING, A LABORATORY MANUAL, (supra). ). In recombinant expression systems in eukaryotes, for the secretion of a protein translated into the lumen of the endoplasmic relic, in the periplasmic space or in the extracellular environment, appropriate secretory signals can be incorporated into the expressed polypeptide. These signals may be endogenous to the polypeptide or may be heterologous signals.

The polypeptides of the present invention can be recovered and purified from recombinant cell culinae through well-known methods including ammonium sulfate or ethanol precipitate, acid extraction, anion exchange or cation chromatography, phosphocellulose chromathography, hydrophobic interaction, affinity chromatography, hydroxylapatty chromatography and lecithin chromalography. Most preferably, ion metal affinity chromatography (IMAC) is used for the purification. Well-known techniques for refolding proteins can be employed to regenerate active confirmation when the polypeptide is denatured during intracellular synthesis, isolation and purification. The expression system can also be a recombinant living microorganism, such as a virus or bacteria. The gene of interest can be inserted into the genome of a recombinant live virus or bacterium. Inoculation and infection in vivo with this live vector will lead to the in vivo expression of the antigen and the induction of immune responses. Viruses and bacteria used for this purpose are, for example: pustulation virus (eg, vaccine, poultry pustulation, canary pustulation), alphavirus (Sindbis virus, Semliki forest virus, Venezuelan equine encephalitis virus), adenovirus, virus associated with adeno, piconavirus (poliovirus, rhinovirus), herpes virus (varicella zoster virus, etc.), Listeria, Salmonella, Shigella, BCG. These viruses and bacteria can be virulent, or pollen in various forms in order to maintain live vaccines. Said live vaccines are also part of the invention.

Diagnostic tests. Prognosis, Serotyping and Mutation This invention also relates to the use of BASB034 polynucleotides and polypeptides of the invention to be used as diagnostic reagents. The detection of polynucleotides and / or BASB034 polypeptides in a eukaryote, particularly a mammal and especially a human being, will provide a diagnostic method for the diagnosis of disease, formation of diseases or response of an infectious organism to drugs. Eukaryotes, particularly mammals, and especially humans, particularly those infected or suspected of being infected with an organism comprising the BASB034 protein gene, can be detected at the nucleic acid or amino acid level through a variety of well-known techniques. known, as well as through methods provided here. The polypeptides and polynucleotides for prognosis, diagnosis and other analyzes can be obtained from bodily ma- terials of infected and / or infected individuals. The polynucleotides of any of these fuenles, parficularly DNA or RNA, can be used directly for defection or. can be amplified enzymatically using PCR or any other amplification technique before analysis. RNA, particularly mRNA, cDNA and genomic DNA can also be used in the same forms. Using amplification, the characterization of the species and strain of an infectious organism or resident present in an individual can be done through an analysis of the genotype of a polynucleotide selected from the organism. The deletions and insertions can be determined by a change in the size of the amplified production compared to a genotype of a reference sequence selected from a related organism, preferably a different species of the same genus or a different strain of the same species. Point mutations can be idenlified by hybridizing DNA by amplifying labeled BASB034 polynucleotide sequences. Perfectly or meaningfully identical sequences can be distinguished from duplexes in perfect or more significantly non-identical by DNase or RNase digestion, for DNA or RNA respeclimamenie, or by de fi ning differences in fusion or cinefication of renafuralization. The polynucleotide sequence differences can also be detected through alterations in the electrophoretic mobility of polynucleotide fragments in gels as compared to a reference sequence. This can be done with or without de-inking agents. Polynucleotide differences can also be determined through direct sequencing of DNA or RNA. See, for example, Myers and others, Science, 230: 1242 (1985). Sequence changes at specific sites can also be revealed through nuclease profiling assays, such as RNase, V1 or S1 protection assay or a chemical cleavage method. See, for example, Cotíon y oíros, Proc. Natl. Acad. Sci. USA, 85 / 4397-4401 (1985). In another embodiment, an array of oligonucleotide probes comprising the nucleoide sequence of BASB034 or its fragments can be constructed to conduct an efficient classification of, for example, genetic mutations, serotype, classification and taxonomic identification. Disposal technology methods are well known and have general applicability and can be used to address a variety of issues in molecular genetics including gene expression, genetic linkage and genetic variability (see, for example, Chee et al., Science, 274). : 610 (1996)). In this manner, in other aspect, the present invention relates to a diagnostic kit comprising: (a) a polynucleotide of the present invention, preferably the nucleotide sequence of SEQ ID NO: 1, 3, 5 or 7, or a fragment of it; (b) a nucleotide sequence complementary to that of (a); (c) a polypeptide of the present invention, preferably the polypeptide of SEQ ID NO: 2, 4, 6 or 8 or a fragment thereof; or (d) an antibody to a polypeptide of the present invention, preferably to the polypeptide of SEQ ID NO: 2, 4, 6 or 8. It will be appreciated that in any of this type of equipment, (a), (b) m (c) or (d) may comprise a substantial component. Said equipment will be used in the diagnosis of a disease or susceptibility to a disease, among others. This invention also relates to the use of polynucleotides of the present invention as diagnostic reagents. The defection of a mulated form of a polynucleotide of the invention, preferably, SEQ ID NO: 1, 3, 5 or 7, which is associated with a disease or pathogenicity, will provide a diagnostic tool that can be added to, or define a diagnosis of a disease, a prognosis of a disease course, a determination of the stage of a disease, or a susceptibility to a disease, which results from subexpression, overexpression or altered expression of the polynucleotide. Organisms, particularly infectious organisms, which carry mulations in said polynucleotide, can be detected at the level of the polynucleotide through a variety of techniques, such as those described herein. The cells of an organism carrying mutations or polymorphisms (allelic variations) in a polynucleotide and / or polypeptide of the invention may also be deduced at the level of polynucleotide or polypeptide through a variety of techniques, to allow the seroxylation, for example. For example, RT-PCR can be used to determine mutations in RNA. It is particularly preferred to use RT-PCR in conjunction with automatic defection systems, such as, for example, GeneScan. RNA, cDNA or genomic DNA can also be used for the same proposyl, PCR. As an example, PCR primers complementary to a polynucleotide encoding the BASB034 polypeptide can be used to identify and analyze mutations. The invention further provides primers with 1, 2, 3, or 4 nucleotides removed from the 5 'and / or 3' exíremo. These initiators can be used, enire you will hear things, to amplify BASB034 DNA and / or RNA isolated from a sample derived from an individual, such as a body material. The primers can be used to amplify a polynucleotide isolated from an infected individual, so that the polynucleotide can then be subjected to various techniques to produce the polynucleotide sequence. In this way, mutations in the polynucleotide sequence can be detected and used for the diagnosis and / or prognosis of the infection or its stage or course, or the serotype and / or classifying the infectious agent. The invention further provides a process for diagnosing, a disease, preferably bacterial infections, most preferably infections caused by Moraxella catarrhalis, which comprises removing from a sample derived from an individual, such as a material of the body, a high level of expression of polynucleotide having a sequence of SEQ ID NO: 1, 3, 5 or 7. The high or decreased expression of a BASB034 polynucleotide can be measured using any of the methods well known in the art for quantification of polynucleotides, such as, for example, amplification , PCR, RT-PCR, RNase protection, Northern staining, spectrometry and other hybridization methods. In addition, a diagnostic assay according to the invention for detecting overexpression of the BASB034 polypeptide, compared to normal control tissue samples, can be used to detect the presence of an infection for example. The assay techniques that can be used to determine the levels of a BASB034 polypeptide, in a sample derived from a host, or as a body material, are well known to those skilled in the art. Said test methods include radioimmunoassays, competitive binding assays, Western staining analysis, antibody sandwich assays, antibody defection and ELISA assays. The polynucleotides of the invention can be used as components of polynucleotide arrays, preferably arrays or high density gratings. These high density arrays are paricularly useful for diagnostic and prognostic purposes. For example, a group of dots each comprising a different gene, and further comprising a polynucleotide or polynucleotides of the invention, can be used to place a probe to identify, such as using hybridization or amplification of nucleic acid, using a probe obtained or derived from a body sample, to determine the presence of a particular polynucleotide sequence or related sequence in an individual. Such presence may indicate the presence of a pathogen, particularly Moraxella catarrhalis, and may be useful for diagnosis and / or prognosis of disease or the course of a disease. A grid comprising a number of variants of the polynucleotide sequence of SEQ ID NO: 1, 3, 5 or 7 is preferred. It is also preferred that it comprises a number of variants of a polynucleotide sequence encoding the polypeptide sequence of SEQ ID NO: 2, 4, 6 or 8.

Antibodies The polypeptides and polynucleotides of the invention or their variants, or cells expressing them, can be used as immunogens to produce nonspecific antibodies for polypeptide or polynucleotide viruses, respectively. In some preferred embodiments of the invention, antibodies to polypeptides or BASB034 polynucleotides are provided. The antibodies generated by the polypeptides or polynucleotides of the invention can be obtained by administering the polypeptides and / or polynucleotides of the invention, or fragments carrying epitopes of either or both, analogs of either or both, or cells expressing either or both, an animal, preferably one that is not human, using rulin protocols. For the preparation of monoclonal antibodies, any technique known in the art that provides antibodies produced through continuous cell line cultures can be used. Examples include various techniques, such as those by Kohler, G. and Milstein, C, Nature 256: 495-497 (1975); Koxbor et al., Immunology Today 4:72 (1983); Cole and you, pg. 77-96 in MONOCLONAL ANTIBODIES AND CANCER THERAPY, Alan R. Liss, Inc. (1985). The techniques for the production of individual chain antibodies (U.S. Patent No. 4,946,778) can be adapted to produce single chain antibodies for polypeptides or polynucleotides of this invention. Also, transgenic mice, and other animals or organisms, such as other mammals, can be used to express humanized antibodies immunospecific to the polypeptides or polynucleotides of the invention. Alternatively, phage display technology can be used to select genes of anibody with binding acfivities to a polypeptide of the invention, either from a repertoire of cow genes and amplified by PCR of human lymphocytes classified or screened for possessing anti -BASB034 or from natural collections (McCafferty et al., (1990), Naíure 348, 552-554; Marks et al., (1992) Biotechnology 10, 779-783). The affinity of these antibodies can also be improved, for example, through chain intermixing (Clackson et al., (1991) Naturel 352: 628). The above-described antibodies can be used to isolate or identify clones expressing the polypeptides or polynucleotides of the invention, to purify polypeptides or polynucleotides by, for example, affinity chromo-ography. In this way, among others, antibodies against the BASB034 polypeptides or BASB034 polynucleotides can be used to bring infections, particularly bacterial infections. Polypeptide variants include equivalent antigenic, epitope or immunological variants that form a particular aspect of this invention. Preferably, the variant antibody is modified to be the least immunogenic in the individual. For example, if the individual is a human being, the antibody most preferably can be "humanized", wherein the region or regions of complementarity determination of the antibody hybridoma derived has been transplanted to a human monoclonal antibody, for example, as described by Jones and others (1986), Nature 325, 522-525 or Tempesí and oíros, (1991) Biotechnology 9, 266-273.

Antagonists and Agonists - Assays and Molecules The polypeptides and polynucleotides of the invention can also be used to determine the binding of small molecule substrates and ligands in, for example, cells, free cell preparations, chemical libraries and natural product mixtures. These substrates and ligands can be substrates and naïve ligands or they can be functional or functional mimetics. See, for example, Coligan and others, Current Protocols in Immunology 1 (2): Chapter 5 (1991). Classification methods can simply measure the binding of a candidate compound to the polypeptide or polynucleotide, or to cells or membranes which carry the polypeptide or polynucleotide, or a profins of fusion of the polypeptide through a direct or indirect label associated with the candidate compound. Allernatively, the classification method may involve competition with a marked competitor. In addition, these classification methods can test whether the candidate compound results in a signal generated by the activation or inhibition of the polypeptide or polynucleotide, using appropriate defecation systems for the cells comprising the polypeptide or polynucleotide. The inhibitors of acíivación generally are analyzed in the presence of a known agony and the effect on the activation through the agonist by the presence of the candidate compound is observed. The constilutively active polypeptide and / or polypeptides and polynucleotides consiIyufiably expressed can be employed in classification methods for agonisces or reverse inhibitors, in the absence of an agonism or inhibitor, testing whether the candidate compound results in the inhibition of the acfivation of the polypeptide or polynucleotide. , according to the case. In addition, classification methods may simply comprise the steps of mixing a candidate compound with a solution containing a polypeptide or polynucleotide of the present invention, to form a mixture, measuring the activity of the polypeptide and / or polynucleotides BASB034 in the mixture and comparing the aclivity of the BASB034 polypeptide and / or polynucleotide of the mixture with a standard. Fusion proteins, such as those made from the Fc portion and BASB034 polypeptide, as described above, can also be used for high throughput screening assays to identify antagonists of the polypeptide of the present invention, as well as phylogenetic polypeptides and / or functionally related (see, D. Benneft and others, J Mol Recogniíion, 8: 52-58 (1995); and K. Johanson and Oíros, J Biol Chem. 270 (16): 9459-9471 (1995)). Polynucleotides, polypeptides and antibodies that bind to and / or infect with a polypeptide of the present invention can also be used to configure classification methods to detect the effect of aggregate compounds on the production of mRNA and / or polypeptide in cells. For example, an ELISA assay can be constructed to measure levels secreted or associated with polypeptide cells using monoclonal and polyclonal antibodies by standard methods known in the art. This can be used to discover agents that can inhibit or improve the production of the polypeptide (also called antagonism or agony, respectively) from cells or tissues conveniently manipulated. The invention also provides a method for classifying compounds to identify those that improve (agonists) or block (antagonisms) the action of polypepides or polynucleotides BASB034, particularly those compounds that are bacteriostatic and / or bactericidal. The method for classifying may involve high production techniques. For example, for the classification of agonists or antagonisms, a syn- thetic reaction mixture, a cellular com- pound, such as a membrane, cell envelope or cell wall, or a preparation of any of this, comprising the BASB034 polypeptide and a sub- strate or labeled ligand of said polypeptide is incubated in the absence or presence of a candidate molecule that can be an agonist or anangonist BASB034. The ability of the candidate molecule to agonize or antagonize the BASB034 polypeptide is reflected in the decreased binding of the labeled ligand or decreased production of the product of said subtraction. The molecules that come together freely, that is, without inducing the effects of the BASB034 polypeptide, are probably very good antagonisms. Molecules that bind well and, as the case may be, raise production output of the subtracter, increase signal transduction or increase the activity of the chemical channel, are agonists. The defection of the speed or level of, as the case may be, production of the product from the substratum, signal transduction or chemical channel activity can be improved by using a report system. Reporting systems that may be useful in this regard include, but are not limited to, colorimetry, marked subsystem bound to the product, a reporter gene that is sensitive to changes in the activity of polynucleotide or BASB034 polypeptide, and binding assays. known in the art. Another example of a BASB034 agonist assay is a competitive assay that combines BASB034 and a potential agonist with BASB034 binding molecules, recombinant BASB034 binding molecules, natural substrates or ligands or substrate or ligand mimics, under conditions appropriate for an assay of competitive inhibition. BASB034 can be labeled, such as through radioactivity or a calorimetric compound, such that the number of BASB034 molecules bound to a binding molecule or converged to production can be accurately deferred to determine the effectiveness of the potential antagonist. Potential antagonists include, among others, small organic molecules, peptides, polypeptides, and antibodies that bind to a polynucleotide and / or polypeptide of the invention and thus inhibit or quench their activity or expression. Potential antagonists may also be small organic molecules, a peptide such as a prolein or closely related antibody that binds the same sites in the binding molecule, such as a binding molecule, without inducing BASB034-induced acfivities, thus avoiding the action or expression of BASB034 polypeptides and / or polynucleotides excluding BASB034 polypeptides and / or polynucleotides from the binding. Potential antagonisms include a small molecule that binds to and occupies the binding site of the polypeptide thus preventing binding to cell binding molecules, so that a normal biological activity is avoided. Examples of small molecules include, but are not limited to, small organic molecules, peptides or peptide-like molecules. Other poisonous anonymias include anisolinide molecules (see, Okano, J. Neruchem, 56: 560 (1991)).; OLIGODEOXYNUCLEOTIDES AS ANTISENSE INHIBITORS OF GENE EXPRESSION, CRC Press, Boca Raion, FL (1988), for a description of these molecules). Preferred poisidic antagonisms include related compounds and variants of BASB034. In a further aspect, the present invention relates to genetically engineered soluble fusion proteins, comprising a polypeptide of the present invention, or a fragment thereof, and several portions of the constant regions of heavy or light chains of immunoglobulins of several subclasses (IgG, IgM, IgA, IgE). The preferred one as immunoglobulin is the constant part of the heavy chain of human IgG, particularly IgG1, where fusion occurs in the hinge region. In a particular embodiment, the Fc part can be removed simply by incorporation of a cleavage sequence, which can be divided with the blood coagulation factor (Xa). Furthermore, the invention relates to processes for the preparation of these fusion proteins through genetic engineering design, and to use same for drug classification, diagnostics and therapy. A further aspect of the invention also discloses polynucleotides encoding said fusion proteins. Examples of fusion protein technology can be found in the patent applications Nos. WO94 / 29458 and WO94 / 22914. Each of the polynucleotide sequences provided herein can be used in the discovery and development of aliphatic compounds. This encoded protein, after expression, can be used as a target for the classification of antibacterial drugs. In addition, the polynucleotide sequences encoding the amino terminal regions of the encoded protein or Shine-Delgarno and other sequences that facilitate translation of the respective mRNA can be used to construct antisense sequences to control the expression of the coding sequence of interest. The invention also provides the use of the polypeptide, polynucleotide, agonist or antagonist of the invention to interfere with the initial physical interaction between a pathogen or pathogens and a eukaryotic, preferably mammalian, host responsible for the sequelae of infection. In particular, the molecules of the invention can be used in: the prevention of adhesion of bacteria, gram-positive and / or gram-negative bacteria to eukaryotes, preferably mammals, extracellular matrix proteins in resident devices or in extracellular maize cells in wounds; for blocking bacterial adhesion between eukaryotic extracellular matrix proteins, preferably mammalian and BASB034 bacterial proteins that mediate the damage of the tumor and / or to block the normal progression of paleogenesis in infections initiated elsewhere through implantation of resident devices or through ofras surgical techniques. According to another aspect of the invention, BASB034 agonists and antagonists are provided, preferably bacteriostatic or bactericidal agonists and antagonists. The anonymiasis and agonists of the invention can be used, for example, to prevent, inhibit and / or eradicate diseases. In a further aspect, the present invention relates to mimotopes of the polypeptide of the invention. A mimotope is a peptide sequence, sufficiently similar to the native peptide (sequentially or structurally), which is capable of being recognized by antibodies that recognize the naive peptide; or is able to develop antibodies that recognize the naive peptide when coupled to a suitable vehicle. The peptide mimophobes can be designed for a particular purpose through the addition, elimination or sub-dilution of selected amino acids. In this manner, the peptides can be modified for the purpose of facilitating conjugation to a protein carrier. For example, it may be desirable for some chemical conjugation methods to include a terminal cisine. further, it may be desirable that the peptides conjugated to a protein poriner include a hydrophobic term away from the conjugate term of the peptide, such that the free unconjugated end of the peptide remains associated with the surface of the carrier protein. In this way, the peptide is presented in a conformation that very closely resembles that of the peptide according to the nafiva molecule found in the background. For example, the peptides can be altered to form an N-terminal cysteine and a C-terminal hydrophobic amidated eximere. Alternatively, the addition or subsiding of a stereoisomer D form of one or more of the amino acids can be performed to create a beneficial derivative, for example, to improve the stability of the peptide. Alternatively, the peptide mimotopes can be idenified using antibodies which are themselves capable of binding to the polypeptides of the present invention using technical techniques such as phage display technology (EP 0 552 267 B1). This technique generates a large number of peptide sequences that resemble the structure of the native peptides and, therefore, are capable of binding to antinative peptide antibodies, but not necessarily by themselves share significant sequence homology to the native peptide. .

Vaccines Other aspect of the invention refers to a method for inducing an immune response in an individual, particularly a mammal, preferably humans, comprising inoculating the individual with the polynucleotide and / or BASB034 polynucleotide, or a fragment or variant thereof. , suitable for producing the antibody and / or a T cell immune response to protect said individual from infection, particularly bacterial infection and very particularly infection by Moraxella catarrhalis. Methods are also provided by which said immunological response decreases bacterial replication. In yet another aspect, the invention relates to a method for inducing immune responses in an individual, comprising supplying said individual with a nucleic acid, sequence or ribozoma neighbor to direct the expression of the BASB034 polynucleotide and / or polypeptide, or a fragment or variant. thereof, to express the polynucleotide and / or BASB034 polypeptide, or a fragment or variant thereof in vivo for the purpose of inducing an immune response, such as to produce an antibody and / or a T cell immune response, including, for example, T-cells for the production of kylosin or cyclo-toxic T-cells, to protect said individual, preferably a human being, from the disease, if that disease is already established within the individual or not. An example for administering the gene is by accelerating it to the desired cells as a cover on particles or in another way. Said nucleic acid vecfor may comprise DNA, RNA, a ribozyme, a modified nucleic acid, a DNA / RNA hybrid, a DNA / proiein complex or an RNA / proiein complex. A further aspect of the invention relates to an immunological composition that when produced to an individual, preferably a human being, capable of inducing an immune response therein, induces an immune response in said individual to a polynucleotide and / or polypeptide. BASB034 encoded therein, wherein the composition comprises a recombinant BASB034 polynucleotide and / or polypeptide encoded therefrom and / or comprises DNA and / or RNA encoding and expressing an antigen of the BASB034 polynucleotide, encoded polypeptide thereof or another polypeptide of the invention . The immunological response can be used therapeutically or prophylactically and can take the form of antibody immunity and / or cellular immunity such as cellular immunity arising from CTL or CD4 + T cells. A BASB034 polypeptide or fragment thereof can be fused to a co-protein or chemical moiety that may or may not produce antibodies, but which is capable of stabilizing the first protein and producing a fused or modified protein that will have antigenic properties and / or immunogenic, and preferably protective properties. In this manner, the fused recombinant protein, further preferably comprises an antigenic co-protein, such as lipoprotein D from Haemophilus inlfuenzae, Glutafiona-S-transferase (GST) or beta-galactosidase, or any other relatively large co-prolein which solubilizes the Prolein and facilitates the production and purification of it. In addition, the co-protein can act as an auxiliary in the sense of providing a generalized stimulation of the immune system of the organism receiving the protein. The co-protein can be linked to any of the amino or carboxy terms of the first protein.

The present invention provides compositions, particularly vaccine compositions, and methods comprising the polypeptides and / or polynucleotides of the invention and immunosfimulatory DNA sequences, such as those described by Safo, Y. et al., Science 273: 352 (1996). Also, the present invention provides methods using the described polynucleotide or its particular fragments, which have been shown to encode non-variable regions of bacterial cell surface proteins., in polynucleotide constructs used in said genetic immunization experiments in animal models of infection with Moraxella catarrhalis. Such experiments will be particularly useful for identifying protein epitopes capable of eliciting a prophylactic or therapeutic immune response. It is believed that this aspect will follow the subsequent preparation of monoclonal antibodies of particular value, derived from the requisite organ of the animal that successfully resists or eliminates the infection, for the development of prophylactic agents or therapeutic treatments of bacterial infection, particularly infection by Moraxella catarrhalis, in mammals, in particular human beings. The invention also includes a vaccine formulation comprising an immunogenic recombinant polypeptide and / or polynucleotide of the invention in June with a suitable vehicle, such as a pharmaceutically acceptable carrier. Since the polypeptides and polynucleotides can be separated in the stomach, each is preferably administered parenterally, including, for example, administration which is subcutaneous, intramuscular, intravenous or intradermal. Formulations suitable for parenteral administration include sterile aqueous and non-aqueous injection solutions which may contain antioxidants, pH regulators, bacteriosylic compounds and solvents which render the formulation isolithic with the body fluid, preferably the blood, of the individual.; and aqueous and non-aqueous sterile suspensions which may include suspending agents or thickening agents. The formulations may be present in single-dose or multi-dose containers, for example, sealed ampoules and flasks and may be stored in a freeze-dried condition that requires only the addition of the sterile liquid vehicle immediately before use. The vaccine formulation of the invention may also include auxiliary systems to improve the immunogenicity of the formulation. Preferably, the auxiliary system arises preferentially to a response type TH1. An immune response can be broadly distinguished into two extreme categories, being a humoral or immune response mediated by cell (traditionally characterized by mechanism of antibody effectors and protective cells, respectively). These response categories have been termed TH1-type responses (cell-mediated response), and TH2-type immune responses (humoral response).

The immune responses of type TH1 exfremas can be characterized by the generation of specific anigen, cytopioxic T lymphocytes restricted to haplotype and natural cell killer response. In mice, TH1-type responses are usually characterized by the generation of IgG2a subtype antibodies, whereas in humans, they correspond to IgG1-like antibodies. The immune responses of lipo TH2 are characterized by the generation of a wide scale of immunoglobulin isotypes including in rails lgG1, IgA and IgM. It can be considered that the activation force beyond the development of these two types of immune responses are the cytosines. High levels of TH1-type cytosines tend to favor the induction of cell-mediated immune responses to the given antigen, while all levels of TH2-type cytosines tend to favor the induction of humoral immune responses to the antigen.

The distinction between immune responses of type TH1 and TH2 is not absolute. In fact, an individual will suppress an immune response that is described as being predominantly TH1 or predominantly TH2. However, it is usually convenient to consider cytosine families in terms of those described in CD4 + ve T cell clones by Mossman and Coffman (Mosmann, TR and Coffman, RL (1989) TH1 and TH2 cells: different patterns of lymphokine secretion lead to different functional properties, Annual Review of Immunology, 7, p145-173). Traditionally, the responses of type TH1 are associated with the production of the cyninsines INF-? and IL-2 through T lymphocytes. Other cytokines usually directly associated with the induction of immune responses of the TH1 type are not produced by T cells, such as IL-12. In coníraste, TH2 type responses are associated with the secretion of IL-4, IL-5, IL-6 and IL-13. It is known that certain vaccine auxiliaries are particularly suitable for the stimulation of cytosine responses of either TH1 or TH2 type. Traditionally, the best indicators of the TH1: TH2 balance of the immune response after a vaccination or infection include direct measurement of the production of TH1 or TH2 type cytosines through T lymphocytes in vitro after restimulation with antigen, and / or the measurement of the IgG1: IgG2a ratio of antigen-specific antibody responses. In this way, a TH1-type helper is one that preferentially stimulates isolated T-cell populations to produce high levels of TH1-type cytosines when they are re-stimulated with antigen in vitro and promotes the development of both CD8 + cytotoxic T lymphocytes and response of immunoglobulin specific antigen associated with the TH1 type isotype. Auxiliaries that are capable of preferential stimulation of the TH1 cell response are described in the international patent application No. WO94 / 00153 and WO95 / 17209. The lipid A of monophosphoryl 3 De-O-acylated (3D-MPL) is one of these auxiliaries. This is known to come from GB 2220211 (Ribi). Chemically, it is a lipid A mixture of 3 De-O-acylated monophosphoryl with 4, 5 or 6 acylated chains and is manufactured by Ribi Immunochem, Montana. A preferred form of 3 De-O-acylated monophosphoryl lipid A is described in European patent 0 689454 B1 (Smihikini Beecham Biologicals SA). Preferably, the 3D-MPL particles are small enough to be sterile filtered through a 0.22 micron window (European patent number 0 689 454). 3D-MPL will be present in the range of 10μg - 100μg, preferably 25-50μg per dose, where the antigen will typically be present on a scale of 2-50μg per dose. Another preferred aid comprises QS21, a non-toxic fraction purified by Hplc derived from the bark of Quillaja Saponaria Molina. Optionally, it can be mixed with 3 De-O-acylated monophosphoryl lipid A (3D-MPL), optionally June with a vehicle. The production method of QS21 is described in the patent of E.U.A. No. 5,057,540. The non-reactogenic auxiliary formulations that confect QS21 have been previously described (WO96 / 33739). Said formulations comprising QS21 and cholesterol have been shown to be useful TH1 stimulant aids when formulated together with an antigen. Other auxiliaries that are preferential stimulants of the TH1 cell response include immunomodulatory oligonucleotides, for example, non-methylated CpG sequences as described in WO 96/02555. Combinations of different TH1 stimulatory aids, such as those mentioned above, are also contemplated by providing an auxiliary which is a preferential stimulant of the TH1 cell response. For example, QS21 can be formulated together with 3D-MPL. The relationship of QS21: 3D-MPL will typically be of the order 1:10 to 10: 1; preferably from 1: 5 to 5: 1 and usually substantially 1: 1. The preferred scale for optimal synergy is 2.5: 1 to 1: 1 of 3D-MPL: QS21. Preferably, a vehicle may also be present in the vaccine composition according to the invention. The vehicle can be an oil-in-water emulsion, or an aluminum salt, such as aluminum phosphate or aluminum hydroxide. A preferred oil-in-water emulsion comprises a metabolizable oil, such as squalene alpha-tocopherol and Tween 80. In a particularly preferred aspect, the antigens in the vaccine composition according to the invention are combined with QS21 and 3D-MPL in said emulsion. In addition, the oil-in-water emulsion may contain span 85 and / or leciin and / or íricaprilin. Typically for human administration, QS21 and 3D-MPL will be present in a vaccine in the range of 1 μg - 200μg, eg as 10-100μg, preferably 10μg 50μg per dose. Typically, the oil in water will comprise 2 to 10% squalene, 2 to 10% alpha-locoferol and 0.3 to 3% tween 80. Preferably, the squalene: alpha-tocopherol ratio is equal to or less than 1 and that this provides a more stable emulsion. Span 85 can also be present at a level of 1%. In some cases, it may be advantageous if the vaccines of the present invention also contain a stabilizer. Non-toxic oil-in-water emulsions preferably contain a non-toxic oil, for example, squalane or squalene, an emulsifier, for example Tween 80, in an aqueous vehicle. The aqueous vehicle can be, for example, saline regulated in its pH with phosphate. A particularly powerful auxiliary formulation involving QS21, 3D-MPL and tocopherol in an oil-in-water emulsion are described in WO 95/17210. The present invention also provides a polyvalent vaccine composition comprising a vaccine formulation of the invention in combination with other animals, in particular antigens useful for bringing about cancers, immune diseases and related conditions. Said polyvalenie vaccine composition may include a TH-1 induction aid as described above. Although the invention has been described with reference to certain BASB034 polypeptides and polynucleotides, it should be understood that it covers fragments of the naturally occurring polypeptides and polynucleotides, and similar polypeptides and polynucleotides with additions, deletions or substi tutions that do not substantially affect the immunogenic properties of the recombinant polypeptides or polynucleotides.

Compositions, Equipment and Administration In a further aspect of the invention, there are provided compositions comprising a BASB034 polynucleotide and / or a BASB034 polypeptide for administration to a cell or a multicellular organism. The invention also relates to compositions comprising a polynucleotide and / or polypeptides discussed herein or their agonists or ani-anagonists. The polypeptides and polynucleotides of the invention may be employed in combination with a non-sterile or sterile vehicle or vehicles for use with cells, tissues or organisms, such as a pharmaceutical carrier suitable for administration to an individual. Said compositions comprise, for example, a media additive or a therapeutically effective amount of a polypepide and / or polynucleotides of the invention, and a pharmaceutically acceptable carrier or excipient. Such vehicles may include, but are not limited to, saline, salt regulated in their pH, dextrose, water, glycerol, ethanol and combinations thereof. The formulation must be adapted to the mode of administration. The invention further relates to diagnostic and pharmaceutical packages and equipment comprising one or more containers filled with one or more of the ingredients of the aforementioned compositions of the invention. Polypeptides can be employed as polynucleotides and other compounds of the invention alone or in combination with other compounds, such as therapeutics. The pharmaceutical compositions may be administered in any effective, convenient manner including, for example, administration via routes topically, orally, anally, vaginally, intravenously, intraperiponeally, intramuscularly, subcutaneously, intranasally or intradermally between ears. In therapy or as a prophylactic, the active agent can be administered to an individual as an injectable composition, for example, as a sterile, preferably isotonic aqueous dispersion. In a further aspect, the present invention provides pharmaceutical compositions comprising a therapeutically effective amount of a polypeptide and / or polynucleotide, ta I as the soluble form of a polypeptide and / or polynucleotide of the present invention, peptide, agonist or antagonist or compound of small molecule, in combination with a pharmaceutically acceptable carrier or excipient. Such vehicles include, but are not limited to, saline, salt regulated in their pH, dextrose, water, glycerol, ethanol and combinations thereof. The invention further relates to pharmaceutical packages and equipment comprising one or more containers filled with one or more of the ingredients of the aforementioned compositions of the invention. The polypeptides, polynucleotides and other compounds of the present invention can be used alone or together with other compounds, as therapeutic compounds. The compositions will be adapted to the route of administration, for example, through a sysiemic or oral route. Preferred forms of systemic administration include injection, typically through intravenous injection. Other routes of injection, such as subcutaneous, intramuscular or intraperitoneal, can be used. Alfernative means for systemic administration include transmucosal and transdermal administration, utilizing penetration agents such as bile salts or fusidic acids or other detergents. In addition, if a polypeptide or other compounds of the present invention can be formulated in an enteric or encapsulated formulation, oral administration is also possible. The administration of composite spheres can also be lopic and / or localized, in the form of balsams, pastes, gels, solutions, powders, and the like. For administration to mammals and particularly humans, it is expected that the daily dose level of the active agent will be 0.01 mg / kg s 20 mg / kg, typically around 1 mg / kg. The doctor will in any case delermine the actual dose that will be the most appropriate for an individual and will vary with the age, weight and response of the particular individual. The above doses are illusive of the average case. In this way they can be individual cases, where scales of doses more aliases or lower are added, and these are within the scope of the invention. The dose scale required depends on the selection of the peptide, the route of administration, the nature of the formulation, the nature of the condition of the subject, and the judgment of the attending physician. However, the appropriate doses are in the range of 0.1-100 μg / kg of the subject. A vaccine composition conveniently is in injectable form. Conventional auxiliaries can be used to improve the immune response. A suitable unit dose for vaccination is 0.5-5 micrograms / kg of the antigen and said dose is preferably administered 1-3 times and in a range of 1-3 weeks. With the indicated dose scale, no adverse toxicological effects will be observed with the compounds of the invention that could prevent their administration to suitable individuals. Wide variations in the dose required, however, are expected to be in view of the variety of compounds available and the different efficiencies of various routes of administration. For example, oral administration is expected to require higher doses than administration by intravenous injection. Variations in these dose levels can be adjusted using standard empirical ruin for optimization, as is well known in the art.

Sequence Databases, Sequences in a Tangible Medium and Algorithms The polypeptide and polynucleotide sequences form a valuable information resource with which one can determine their 2- and 3- dimensional structures as well as identify other sequences of similar homology. These aspects are greatly facilitated by storing the sequence in a computer-readable medium and then using the data stored in a program of known macromolecular structure or to search for a sequence-based database using well-known search tools, such as the GCG program package. . The invention also provides methods for the analysis of character sequences or chains, particularly genetic sequences or encoded protein sequences. Preferred methods of analysis include, for example, sequence homology analysis methods, such as identity and similarity analysis, DNA, RNA, and protein structure analysis, sequence assembly, cladistic analysis, sequence motif analysis, determination of reading frame, called nucleic acid base, codon usage analysis, nucleic acid base separation and peak analysis of sequencing chromatogram. A computer-based method is provided to perform homology identification. This method comprises the steps of: providing a first polynucleotide sequence comprising the sequence of a polynucleotide of the invention in a computer-readable medium; and comparing the first polynucleotide sequence with at least one second polynucleide or polypeptide sequence to idenify the homology. A computer-based method may also be provided to perform homology identification, the method comprising the steps of: providing a first polypeptide sequence comprising the sequence of a polypeptide of the invention in a computer-readable medium; and comparing the first polypeptide sequence with at least one second polynucleotide or polypeptide sequence to identify the homology. All publications and references, including, but not limited to, patents and patent applications, cited in this specification are incorporated herein by reference in their entirety, as if each publication or individual reference was specifically and individually indicated as being incorporated herein by reference. as it has been fully established. Any patent application which is claim priority claim is also incorporated by reference herein in its entirety in the manner described above for publications and references.

DEFINITIONS "Identity", as is known in the art, is the relationship between two or more polypeptide sequences or two or more polynucleotide sequences, as the case may be, as determined by comparing the sequences. In the art, "identity" also represents the degree of sequence relationship between polypeptide and polynucleotide sequences, as the case may be, as determined by the coincidence of strips of said sequences. The "identity" can be easily calculated by known methods, including, but not limited to, those described in Computational Molecular Biology, Lesk, A.M., ed., Oxford Universily Press, New York, 1988; Biocomputing; Informatics and Genome Projects, Smith, D. W., ed. Academic Press, New York, 1993; Computer Analysis of Sequence Data, Part I, Griffin, A.M., and Griffin, H.G., eds., Humana Press, New Jersey, 1994; Sequence Analysis in Molecular Biology, von Heine, G., Academic Press, 1987; and Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M Stockfon Press, New York, 1991; and Carillo, H., and Lipman, D., SIAM J. Applied Math, 48: 1073 (1988). Methods have been designed to determine the identity to give the greatest match between the tested sequences. In addition, methods to determine identity are encoded in publicly available computer programs. The computer program methods for determining the identity of between two and sequences include, but were not limited to the GAP program in the GCG program package (Devereux, J. et al., Nucleic Acids Research 12 (1): 387 (1984)), BLAST, BLASTN (Alfschul, SF and others, J. Molec. Biol. 2l5 / 403-410 (1990), and FASTA (Pearson and Lipman Proc. Natl. Acad. Sci. USA 85, 2444-2448 (1988) .The BLAST family of programs is publicly available from NCBI and other sources (BLAST. Manual, Altschul, S. et al., NCBI NLM NIH Bethseda, MD 20894; Atlschul, S., and others, J. Mol. Biol. 21: 403-410 (1990). It can also be used to determine identity, the well-known Smith Waferman algorithm.

The parameters for polypeptide sequence comparison include the following: Algorithm: Needleman and Wunsch, J. Mol. Biol. 48: 443-453 (1970) Comparison matrix: BLOSSUM62 by Henikoff and Henikoff, Proc. Natl. Acad. Sci. USA. 89: 10915-10919 (1992) Gap penalty: 8 Gap length penalty: 2 A useful program with these parameters is publicly available as the "gap" program of Genetics Computer Group, Madison Wl. The aforementioned parameters are the default parameters of peptide comparisons (along with no penalty for voids). The parameters for the polynucleotide comparison include the following: Algorilmo: Needleman and Wunsch, J. Mol. Biol. 48: 443-453 (1970) Comparison maiz: coincident = +10, mismatch = 0 Gap penalty: 50 Gap length penalty: 3 Available as: The gap program of Genetics Compuer Group, Madison Wl. These are the default parameters for nucleic acid comparisons. A preferred meaning for "identity" for polynucleotides and polypeptides, as the case may be, are provided in (1) and (2) below. (1) Polynucleotide embodiments further include an isolated polynucleotide comprising a polynucleotide sequence having at least one identity of 50, 60, 70, 80, 85, 90, 95, 97, 0 100% > to the reference sequence of SEQ ID. NO: 1, wherein said polynucleotide sequence may be identical to the reference sequence of SEQ ID NO: 1, or may include up to a number of nucleotide alions as compared to the reference sequence, wherein such alterations are selected of the group consisting of at least one deletion, subsliution, including ransition and ransversion, or nucleotide insertion, and in such alleles may occur at the 5 'or 3'-iesminal positions of the reference nucleotide sequence or in any part between those terminal positions, interspersed either individually between the nucleotides in the reference sequence or in one or more contiguous groups within the reference sequence, and wherein number of nucleotide alterations is determined by multiplying the tofal number of nucleoides in SEC ID NO: 1, by the number that defines the percentage of identity divided by 100 and then subtracts the produlum of the total number of nucleotides in SEQ ID NO: 1, or: nn < xp - (xn • y), where, nn is the number of nucleotide alferations, xn is the total number of nucleotides in SEQ ID NO: 1, and is (0.50 for 50%, 0.60 for 60%, 0.70 for 70%, 0.80 for 80%, 0.85 for 85%, 0.90 for 90%, 0.95 for 95% 0.97 for 97% or 1.00 for 100%, and • is the symbol for the multiplication operator, and where any product that is not integer xn and e is rounded down to the nearest whole before subtracting it from xn Alterations of a polynucleotide sequence encoding the polypeptide of SEQ ID NO: 2 can create non-sense, missing sense or frame shift mutations in this coding sequence and in this way they alter the polypeptide encoded by the polynucleotide followed by said alterations For example, a polynucleotide sequence of the present invention will be identical to the reference sequence of SEQ ID NO: 1, which can be 100% identical, or can include up to number number of nucleic acid alterations as compared to the reference sequence, so that the percent identity is less than 100% identity. Said alterations are selected from the group consisting of at least one deletion, substitution, including transition and transversion, or nucleic acid insertion, and wherein alterations may occur at the 5 'p 3' terminal positions of the polynucleotide sequence of reference or anywhere between those terminal positions, interspersed either individually between the nucleic acids in the reference sequence or in one or more contiguous groups within the reference sequence. The number of nucleic acid alterations for a given percentage of identity is determined by multiplying the nucleic acid number in SEQ ID NO: 1 by the integer which defines the percent identity divided by 100 and then subtracting that product from the total number of acids nucleic acids in SEQ ID NO: 1, or: nn < xn - (x "• y), wherein, nn is the number of nucleic acid alterations, xn is the tofal number of nucleic acids in SEQ ID NO: 1, and is, for example, 0.70 for 70%, 0.80 for 80%, 0.85 for 85%, etc. ., • is the symbol for the multiplication operator, and where any product that does not integer from xn and y is rounded down to the nearest integer before subtracting it from xn. (2) Additional polypeptide embodiments include an isolated polypeptide comprising a polypeptide that is at least 50, 60, 70, 80, 85, 90, 95, 97, or 100% identity for a polypeptide reference sequence. of SEC ID. NO: 2, wherein said polypeptide sequence may be identical to the reference sequence of SEQ ID NO: 2, or may include a close number of amino acid alterations as compared to the reference sequence, wherein said alterations are selected of the group consisting of at least one deletion, substilution, including conservative and non-conservative substitution, or nucleic acid insertion, and wherein such alterations may occur at the amino- or carboxy-terminal positions of the reference polypeptide sequence anywhere between those terminal positions, interspersed either individually between the amino acids in the reference sequence or in one or more contiguous groups, with the reference sequence, and where number of amino acid alterations is determined by multiplying the total number of amino acids in SEQ ID NO: 2, by the whole defining the percentage of identity divided by 100 and after s subtracting that product from toíal number of amino acids in SEQ ID NO: 2, or: na < xa - (xa • y), where, na is the number of amino acid aliases, xa is the photo number of amino acids in SEQ ID NO: 2, and is (0.50 for 50%, 0.60 for 60%, 0.70 for 70%, 0.80 for 80%, 0.85 for 85%, 0.90 for 90%, 0.95 for 95% 0.97 for 97% or 1.00 for 100%, and • is the symbol for the multiplication operator, and where any product that does not whole from x to y is rounded down to closest integer before substrating it from xa By way of example, the polypeptide sequence of the present invention may be identical to the reference sequence of SEQ ID NO: 2, which may be 100% identical, or may include up to a certain number The amino acid alignments are compared to the reference sequence, so that the identity percentage is less than 100% identity.These alterations are selected from the group consisting of at least one deletion, substifution, including conservative substifution and no conservative, or ami insertion noacid, and where alterations may occur at the amino or carboxy terminal positions of the reference polypeptide sequence or anywhere between those terminal positions, interspersed either individually with the amino acids in the reference sequence or in one or more contiguous groups within the reference sequence. The number of amino acid aliases for an identity percent is determined by multiplying the total number of amino acids in SEQ ID NO: 2, by the integer defining the percentage of identity divided by 100 and then subtracting that product from the total number of amino acids in SEQ ID. NO: 2, or: na < xa - (xa • y), where, na is the number of amino acid aliases, xa is the tofal number of amino acids in SEQ ID NO: 2, and is, for example, 0.70 for 70%, 0.80 for 80%, 0.85 for 85%, etc. , and • is the symbol for the multiplication operator, and where any non-energetic production of xa and y is rounded down to the nearest star before subsuming it from xa. The "individuals", when used in the present with reference to an organism, means a multi-cellular eukary, including, but not limited to, a metazoan, a mammal, an ovipositor, a bovid, an ape, a primate, and a creature. human. "Isolated" means altered "by the hand of man" of his nalural state, that is, if it occurs by nature, has been changed and removed from its original environment or both. For example, a polynucleolide or a natural polypeptide present in a living organism is not "isolated", but the same polynucleotide or polypeptide separated from the coexisting cells of its naíural state is "isolated", as the term is used herein. In addition, a polynucleide or polypeptide that is introduced into an organism through transformation, genetic manipulation or any other recombinant method is "isolated" even if it is still present in another organism, this organism can be living or non-living. The "polynucleotides" generally refer to polyribonucleotide or polydeoxyribonucleotide, which may be unmodified RNA or DNA or modified RNA or DNA, including regions of double-stranded structure, and singular. "Variant" refers to a polynucleotide or polypeptide that differs from a reference polynucleotide or polypeptide, but retains its essential properties. A typical variant of a polynucleotide differs in nucleotide sequence from another reference polynucleotide. Changes in the nucleotide sequence of the variant may or may not alter the amino acid sequence of a polypeptide encoded by the reference polypeptide. The nucleotide changes may result in or amino acid substitutions, additions, deletions, fusions and truncations in the polypeptide encoded by the reference sequence, as discussed below. A typical variant of a polypeptide differs in the amino acid sequence of another reference polypeptide. Generally, the differences are limited such that the sequence of the reference polypeptide and the variant are closely similar in total and, in many regions, identical. A reference variant and polypeptide may differ in an amino acid sequence by one or more substitutions, additions, deletions, in any combination. A substituted or inserted amino acid residue may or may not be one encoded by the genetic code. A variant of a polynucleotide or polypeptide can be of natural existence, as an allelic variant, or it can be a variant that is not known to be of naive existence. Varianies that are not naturally occurring in polynucleolides and polypeptides can be made through mulagenesis techniques or through direct synthesis. "Disease (s)" means any disease caused by or related to infection by a bacterium, including, for example, otiíis media in children and infants, pneumonia in genie major, sinusitis, nosocomal infections, and invasive diseases, chronic otilis media with loss of the ear, fluid accumulation in the middle ear, damage to the auditory environment, slow learning to speak, infection of the upper respiratory tract and inflammation of the middle ear.

EXAMPLES The following examples are made using standard techniques, which are well known to those experienced in the technical field, except when describing what is heard with de alle ue. The examples are illusory, but do not limit the invention.

Example 1: Discovery and sequencing of DNA confirming the BASB034 gene from Moraxella catarrhalis strain ATCC 43617 The BASB034 gene was first discovered in the Incyte PathoSeq database containing unfinished genomic DNA sequences from Moraxella catarrhalis strain ATCC 43617 (also referred to as strain MC2931). The translation of the BASB034 polynucleotide sequence showed an important similarity (42% identity in an overlap of 214 amino acids) to an outer membrane phospholipase A protein Klefsiella pneumoniae. The sequence of the BASB034 gene was also confirmed experimentally. For this purpose, genomic DNA was extracted from 1010 cells of the M. catarrhalis cells (strain ATCC 43617) using the QIAGEN genomic DNA extraction equipment (Qiagen Gmgh), and 1 μg of this material was subjected to DNA amplification of the DNA reaction. polymerase chain using the primers E481124 (5'-GAT TTA AGA GTA TGT TAT GAT G-3 ') [SEQ ID NO: P] and E481125 (5'-GTA TGG GTT GAT CAA ATA CAG-3') [SEQ ID NO: 10] . This PCR product was purified on a Biorobot 9600 (Qiagen GmbH) and subjected to DNA sequencing using the Big Dye Cycle Sequencing kit (Perkin-Elmer) and an ABI 377 / PRISM DNA sequencer. DNA sequencing was performed on both chain strands with a redundancy of 2 and the full length sequence was assembled using Sequencher ™ software (Applied Biosystems). The resulting DNA sequence became 100% identical to SEQ ID NO: 1.

Example 2 Variability analysis of the BASB034 gene between several strains of Moraxella catarrhalis. 2A: Restriction Fragment Length Analysis (RFLP). Genomic DNA was extracted from 16 M. catarrhalis strains (presented in Table 1) as described further below. M. catarrhalis was streaked for individual clones on BHI plates and developed overnight at 37 ° C. Three or four individual colonies were collected and used to inoculate a BHI broth seed culture of approximately 1.5 ml (brain-heart infusion) that was grown overnight in a shaking incubator, approximately 300 rpm at 37 ° C. A 500 ml Erlenmeyer matras containing approximately 150 ml of the BHI broth was inoculated with the seed culture and allowed to develop for approximately 12-16 hours at 37 ° C in a shaking incubator, approximately 175 rpm, to generate cell mass for the isolation of DNA. The cells were collected by centrifugation in a Sorvall GSA rotor at approximately 2000 Xg for 15 min. At ambient temperature. The supernatant was removed and the cell pellet was suspended in approximately 5.0 ml of sterile water. An equal volume of the lysis pH regulator (200 mM NaCl, 20 mM EDTA, 40 mM Tris-HCl, pH 8.0, 0.5% (w / v) SDS, 0.5% (v / v) 2-mercaptoefanol, and 250 μg / ml prokinase K) was added and the cells were suspended through moderate agitation and fitulation. The cell suspension was then incubated approximately 12 hours at 50 ° C to use the bacteria and release chromosomal DNA. The major protein was precipitated through the addition of 0.5 ml of saturated NaCl (approximately 6.0 M in sterile water) and centrifugation at approximately 5,500 xg in a sorvall SS34 rotor at ambient temperature. Chromosomal DNA was precipitated from the clarified supernatant through the addition of two volumes of 100% ethanol. The added DNA was collected and washed using moderate agitation in a small volume of a 70% ethanol solution. The purified chromosomal DNA was suspended in sterile water and allowed to dissolve / combine overnight at 4 ° C through moderate oscillation. The concentration of the dissolved DNA was determined spectrophotomelically at 260 nm using an exclusion coefficient of 1.0 or O.D. unit approximately 50 μg / ml. This material was then subjected to amplification by PCR using oligogonotides MC-Pla-BamF (5 '- AAG GGC CCA ATT ACG CAG AGG GGA TCC CAA GCT GTA CCA AAT CCT GTG GCA TTT GTT G -3') [SEQ ID NO: 11) and MC-Pla-SalRC (5'-AAG GGC CCA ATT ACG CAG AGG GTC GAC TTA TTA TAG ACC CAT CCA GTC GTT AAG CAT AAG -3 ') [SEQ ID NO: 12]. The amplicons of the corresponding BASB034 gene were then subjected to hydrolysis independently using resuscitation enzymes (Hphl)., Alul, Rsa, EcoRV, Sau3A1) and the resiriction products were separated through agarose or polyacrylamide gel electrophoresis using standard molecular biology procedures as described in "Molecular Cloning, a Laboratory Manual, Second Edition, Eds .: Sambrook, Fristch &; Maniatis, Cold Spring Harbor Press 1989. "The photographs of the resulting electrophoresis gels are presented in Figure 1. For each strain, the RFLP patterns corresponding to 6 restriction enzymes were classified and combined.The groups of strains that share a combination Identical RFLP patterns were then identified.Using this meioticology, the strains tested in this study fall into fres genomic groups (Group 1, Mc2931, Ms2904, Mc2905, Mc2907, Mc2909, Mc2910, Mc2911, Mc2912, Mc2926, Mc2931, C2956, Mc2969, Mc2975, Group 2: Mc2906, Mc2913, Group 3: Mc2908. (Mc2960 failed to be amplified and consequently was not classified.) These data support that the population of Moraxella catarrhalis used in this study reveals a limited diversity of nucleotide sequence. for the BASB034 gene. 2B: DNA sequencing in other strains. Using the experimental procedure described in the Example, the BASB034 gene sequence is also determined for the three additional strains of Moraxella catarrhalis. The nucleotide sequences of the BASB034 gene of strains Mc2908, Mc2913 and Mc2969 representative of the three genomic groups identified previously, are shown in SEQ ID NO: 3, 5 and 7, respectively. These nucleophil sequences were translated into amino acid sequences, which are shown in SEQ ID NO: 4, 6 and 8 respectively. Using the MegAlign program of the DNASTAR Lasergene package, a multiple alignment of nucleophilic sequences SEQ ID NO: 1, 3, 5 and 7 was performed and shown in Figure 2. A comparison in identity pairs is summarized in the table 2, showing that the four BASB034 nucleophilic gene sequences are similar to a level of identity greater than 98%. Using the same program, a multiple alignment of the protein sequences of SEQ ID NO: 2, 4, 6 and 8 was performed, and is presented in Figure 3. A comparison in identity pairs is summarized in Table 3, showing that the 4 BASB034 protein sequences are similar at the level of idenity and greater than 98%. In conjunction, these data indicate a very strong sequence conservation of the BASB034 gene between strains of Moraxella catarrhalis.

TABLE 1 Characteristics of the Moraxella catarrhalis strains used in this study TABLE 2 Identities in pairs of BASB034 polynucleotide sequences (in%) TABLE 3 Identities in pairs of BASB034 polynucleotide sequences (in%) EXAMPLE 3 Construction of Plasmid for Expressing Recombinant BASB034 A: Cloning of BASB034 The SamHI and Sa / I restriction sites engineered into the forward amplification primers (SEQ ID NO: 11) and reverse (SEWC ID NO: 12)} , respectively, allowed the directional cloning of a PCR product of BASB034 in the commercially available E. coli expression plasmid pQE30 (QiaGen, resistant to ampicillin), so that a mature BASB034 protein can be expressed as a fusion protein containing an affinity chromatography (His) 6 label in the N-terminus. The PCR product of BASB034 was purified from the amplification reaction using a spin column based on silica gel (QiaGen) according to the instructions of the manufacturers. To produce the required SamHI and Sa / I terms necessary for cloning, the purified PCR product was sequentially digested to complete with the BamHl and Sali resynchronization enzymes as recommended by the manufacturer (Life Techonologies). After the first restriction digestion, the PCR product was purified through the spin column as previously done to remove the salts and eluted in spherical water before the second enzyme digestion. The digested DNA fragment was again purified using spin columns based on silica gel before ligation with the plasmid pQE30.

B: Production of Expression Vector To prepare the expression plasmid pQE30 for ligation, it was similarly digested to complete tannin with BamHl and Sa / I and then it was treated with calf intestinal phosphatase (CIP, at approximately 0.02 units / pmol end). 5 ', Life Technologies) in accordance with the manufacturer's instructions to avoid auloligation. A molar excess of approximately 5 times the digested fragment for the prepared neighbor was used to program the ligation reaction. A ligation reaction of approximately 20μl standard (approximately 16 ° C, approximately 16 hours, using methods well known in the field, was performed using T4 DNA ligase (approximately 2.0 units / Life Technologies reaction). An aliquot of the ligation was used (about 5μl) to transform M15 cells (pREP4) according to methods well known in the art.After a period of overgrowth of about 2-3 hours at 37 ° in about 1.0 ml of LB broth the transformed cells were placed on LB agar plates containing kanamycin (50μl / ml) and ampicillin (100μl / ml) Both antibiotics were included in the selection media to ensure that all transformed cells will carry both the pREP4 (KnR) plasmid, which carries the laclq gene necessary for the repression for the IPTG-inducible expression of proteins in pQE30, and the plasmid pQE30-BASB034 (ApR). The plates were incubated overnight at 37 ° C for about 16 hours. Individual colonies of KnR / ApR were collected with sterile sticks and used to "cover with patches" the KbR / ApR plates of fresh LB inoculated as well as approximately 1.0 ml of KnI / ApR stock of LB broth. Both the patch plates and the broth culm were incubated overnight at 37 ° C in a standard incubator (plate) or in a shaking water bath. A whole-cell-based PCR analysis was used to verify that the transformants contained the BASB034 DNA insert. Here, the overnight LB Kn / Ap broth culture of approximately 1.0 ml was transferred to a 1.5 ml polypropylene tube and the cells were collected by centrifugation in a Beckman microcentrifuge (approximately 3 minutes, ambient temperature, approx. 12,000 X g). The cell pellet was suspended in approximately 200μl of sterile water and an aliquot of approximately 10μl was used to program a PCR reaction with a final volume of approximately 50μl containing the forward amplification primers as inverse BASB034. The final concentrations of the PCR reaction components were essentially the same as those specified in the example, except that approximately 5.0 Taq polymerase was used. The initial 95 ° C denaillization step was increased to 3 min. To ensure thermal breakdown of the bacterial cells and release of the plasmid DNA. A model 9700 ABI thermal cyclizer and a three step, 32 cycle thermal amplification profile, ie, 95 ° C, 45sec, were used.; 55-58 ° C, 45sec, 72 ° C, 1min., To amplify the BASB034 PCR method of the lysate-transforming samples. After the laser amplification, an aliquot of ~ 20μl of the reaction was analyzed through agarose gel electrophoresis (0.8% agarose in a pH regulator of Tris-acephate-EDTA (TAE)). The DNA fragments were visualized through UV illumination after gel electrophoresis and staining with iodide bromide. A DNA molecular size standard (1 Kb size, Life Technologies) was elecrofrown in parallel with the test sample, and used to estimate the size of the PCR products. The transformants that produced the expected PCR product were identified as strains containing a construction of the BASB034 expression. The strains containing the expression plasmid were then analyzed for expression capable of induction of the recombinant BASB034.

C: Expression Analysis of Positive Transformants to PCR For each PCR positive transformant identified above, approximately 5.0 ml of the LB broth was inoculated with kanamycin (50 μg / ml) and ampicillin (100 μg / ml) with cells from the patch plate and grown overnight 37 ° C with agitation (-250 rpm). An aliquium of the nocturnal seedling cullivo (-1.0 ml) was inoculated into a 125 ml Erlenmeyer flask containing approximately 25 ml of the LB Kn / Ap broth and developed at 37 ° C with agitation (-250 rpm) until the ripening of the crop reached an OD 600 of approximately 0.5, that is, average registration phase (usually around 1.5-2.0 hours). At that time, approximately half of the culture (~ 12.5 ml) was transferred to a second 125 ml flask and the expression of the BASB034 protein recombined induced by the addition of IPTG (1.0 M abasification solution prepared in sterile water, Sigma) at a final concentration of 1.0 mM. Incubation of the cultures tanfo induced by IPTG as non-induced continued for approximately 4 additional hours at 37 ° C with shaking. Samples (approximately 1.0 ml) were removed from induced and uninduced cullives after the induction period and the cells were collected by centrifugation in a microcentrifuge at ambient temperature approximately 3 min. The individual cell pellets were suspended in ~ 50μl of sterile water, after mixing with an equal volume of a pH regulator from the 2X Laemelli SDS-PAGE sample containing 2-mercaptoefanol, and placed in a boiling water bath for approximately 3 minutes to de-neutralize the protein. Equal volumes (~ 5μl) of the cell lysates induced with IPGT as uninduced crude were loaded in duplicate in 12% or glycine polyacrylamide tris-gel (minigels with a thickness of 1 mm, Novex). The lisal samples induced as uninduced were elecroformed together with pre-stained molecular weight markers (see Blue, Novex) under conventional conditions by monitoring a standard running pH of SDS / Tris / glycine (BioRad). After electrophoresis, a gel was stained with commassie R250 brillanfe blue (BioRad) and then stained to visualize the novel inducible IPTG media of BASB034. The second gel electroengineed in a PVDF membrane (0.45 micron pore size, Novex) lasts approximately 2 hours at 4 ° C using a BioRad Mini-Protean II ion and Towbin methanol transfer pH regulator (20%) ). The blocking of the membrane and antibody incubations were carried out according to the methods well known in the art. An anti-RGS (His) 3 monoclonal antibody, followed by a second conjugated rabbit ani-mouse antibody for HRP (QiaGen) was used to confirm the expression and identity of the BASB034 recombinant protein. Visualization of the anti-His antibody reagent pattern was achieved using either an insoluble ABT substrate or using a hyperfilm with the Amersham ECL chemiluminescence system.

D: Sequence Confirmation To additionally verify that PROBINE BASB034 recombines inducible mediated IPTG being expressed in the correct open reading frame and is not a spurious molecule arising from a cloning arp (ie, a frame-shift), the DNA sequence of the cloned insert was determined. The DNA sequence for the BASB034 gene of M. catarrhalis was obtained from a chain transfer using conventional asymmetric PCR cycle sequencing methodologies (ABI Prism Dye-Finishing Cycle Sequencing, Perkin-Elmer). Sequencing reactions were programmed with undigested expression plasmid DNA (~ 0.5μg / rxn) as a template and specific sequencing primers in the pQE30 vector and specific in appropriate ORFs (-3.5 pmol / rxn). In addition to the template and the sequencing primer, each sequencing reaction (~ 20μl) contained the 4 different dNTPs (ie, A, G, C, and T) and the 4 ddNTPs (ie, ddA, ddG, ddC and ddT) corresponding terminator nucleotides; with each terminator being conjugated to one of the four fluorescent colorani, Joe, Tam, Rox or Fam. The individual chain structure sequencing elongation products were determined in random positions throughout the template through the incorporation of the ddNTP eterminers marked with dye. Fermentation products labeled with fluorescent dye were purified using microcentrifuge size exclusion chromatography columns (Princeton Genetics), dried under vacuum, suspended in a resuspension pH buffer template (Perkin-Elmer) for capillary electrosphoresis or Deionized formamide for PAGE, were denatured at 95 ° C for approximately 5 minutes, and analyzed with capillary elecirophoresis of alia resolution (ABI 310 Auíomaíed DNA Sequenator, Perkin-Elmer), or alpha resolution PAGE (ABI 377 Automated DNA Sequenator) according to recommended by the manufacturer. The DNA sequence data produced from individual reactions were collected and the relative fluorescent peak intensities were automatically analyzed on a PowerMAC computer using the software ABI Seqúense Software Analysis (Perkin-Elmer). Individually autoanalyzed DNA sequences were manually edited for accuracy upon being taken to a consensus strand structure sequence "lira" using AutoAssembler Software software (Perkin-Elmer). Sequencing determined that the expression plasmid contained the correct sequence in the correct open reading frame.

Example 4 Production of Recombinant BASB034 Bacterial strain A recombinant expression strain of E. coli M15 (pREP4) containing a plasmid (pQE30) encoding BASB034 of M. catarrhalis was used to produce cell mass for the purification of recombinant protein. The expression strain was cultured on LB agar plates containing 50μg / ml kanamycin ("Kn") and 100μg / ml ampicillin ("Ap") to ensure the conírol plasmid pREP4 laclq as the expression construct pQE30-BASB034 that both were maintained. For cryopreservation at -80 ° C, the strain was propagated in the LB broth containing the same concentration of antibiotics and then mixed with an equal volume of LB broth containing 30% glycerol (w / v).

Media The fermentation medium used for the production of recombinant protein consisted of 2X YT broth (Difco) containing 50μg / ml Kn and 100μg / ml Ap. Antifoam was added to the medium for the fermenter at 0.25ml / L (Aníifoam 204, Sigma). To induce the expression of the BASB034 recombinant protein, IPTG (isopropyl β-D-thiogalacyopyranoside) was added to the fermenter (1 mM, final).

Fermentation A 500 ml Erlenmeyer seed flask, containing 50 ml of work volume, was inoculated with 0.3 ml of rapidly thawed frozen culture, or several colonies of a selective agar plate culture, and incubated for approximately 12 hours at 37 ± 1 ° C on a stirring platform at 150 rpm (Innova 2100, New Brunswick Scientific). The sowing cycle was then used to inoculate a 5 liter working volume fermentor containing the 2X YT broth and the Tanfo Kn antibiotics as Ap. The fermenter (Bioflo 3000, New Brunswick Scientific) was operated at 37 ± 1 ° C, 0.2-0.4 VVM of air spray, 250 rpm in Rushton impellers. The pH value was not controlled either in the flask seed culture or in the fermentor. During fermentation, the pH value of 6.5 to 7.3 in the fermenter. IPTG (1.0 M supply material, prepared in sterile water) was added to the fermenter when the cull reached an average growth record (-0.7 O.D. 600 units). Cells were induced for 2-4 hours, then harvested through centrifugation using either a 28RS Heraeus super-speed centrifuge (Sepatech) or RC5C (Sorval Insfruments). The passage of cells was stored at -20 ° C and had to be processed.

Purification Chemicals and Materials Imidazole, guanidine hydrochloride, Tris (hydroxymethyl) and EDTA (ethylene-diamine tetraacetic acid), of biotechnological grade or better, were all obtained from Ameresco Chemical, Solon, Ohio. Triton X-100 (t-oclilfenoxiplietoxi-anol), sodium phosphate, monobasic and urea were reactive or better grade and were obtained from Sigma Chemical Company, St. Louis, Missouri. Glacial acetic acid and hydrochloric acid were obtained from Mallincrodt Baker, Inc. Phillipsburg, New Jersey. Methanol was obtained from Fisher Scientific, Fairlawn, New Jersey. Pefabloc®SC (4- (2-aminoethyl) -benzensulfonyl fluoride). The proiease complelas inhibitor cochlea and PMSF (phenylmethyl sulfonyl fluoride) tablets were obtained from Roche Diagnostics Corporation, Indianapolis, Indiana. The bestalin, pepstaine A, and protease inhibitor E-64 were obtained from Calbiochem, LaJolla, California. Saline regulated at its pH with dulbeco phosphate (1x PBS) was obtained from Quality Biological Inc., Gaithersburg, Maryland. Saline regulated at its pH with dulbeco phosphate (10x PBS) was obtained from Bio Whittaker, Walkersville, Maryland. The penta-His antibody, free BSA was obfuscated from QiaGen, Valencia, California. AffiniPure goat anti-mouse IgG conjugated with peroxidase was obtained from Jackson Immuno Research, West Grove, Penn. The individual AEC solution was obtained from Zymed, South San Francisco, California. All other chemicals were reactive or better. The Ni-NTA Superflow resin was obtained from QiaGen Inc., Valencia, California. The pre-cast Tris-glycine 4-20% and 10-20% polyacrylamide gels, all run-in pH regulators and solutions, SeeBlue pre-staining standards, MuliiMark multiple color standards and PVDF transfer membranes were obtained from Novex, San Diego, California. Silver SDS-PAGE staining kits were obtained from Daiichi Pure Chemicals Company Limited, Tokyo, Japan. The Coomassie staining solution was obtained from Bio-Rad Laboratories, Hercules, California. Acrodisc® 0.2m syringe films were obtained from Pall Gelman Sciences, Ann Arbor, Michigan. GD / X 25 mm disposable syringe filters were obtained from Whatman Inc., Clifton, New Jersey. The 8000 MWCO dialysis line was obtained from BioDesign Inc. Od New York, Carmal New York. BCA proiein assay reagents and snake skin dialysis tubing 3,500 MWCO were obtained from Pierce Chemical Co., Rockford, Illinois.

Extraction Protocol The cell paste was thawed at room temperature for 30 to 60 minutes. Five to six grams of the material were loaded into a 50 ml disposable centrifuge tube. To these five ml / grams of guanidine hydrochloride (Gu-HCl) was added pH buffer (6M guanidine hydrochloride, 100 mM sodium phosphate, monobasic, 10 mM Tris and 0.05% X-100, pH 8.0 ). The cell pass was resuspended using a PRO300D procyanific homogenizer at an energy of 3/4 for one minute. The exudation mixture was then placed at ambient temperature with moderate agitation for 60 to 90 minutes. After 60 to 90 minutes, the extraction mixture was centrifuged at 15,800 x g for 15 minutes (Sorvall RC5C centrifuge, 11,500 rpm). The supernatant (S1) was decanted and saved for further purification. The pellet (P1) was saved for analysis.

Binding of BASB034 to the nickel-NTA resin To the supernatant S1 were added from 3 to 4 ml of Ni-NTA. This was then placed at ambient temperature with moderate agitation in 1 hour. After one hour, the S1 / NÍ-NTA was packed into a column of XK16 Pharmacia. The column was then washed with 1M Gu-HCl pH buffer (1 M guanidine hydrochloride, 100 mM sodium phosphate, monobasic, 10 mM Tris and 0.05% triton X-100, pH 8.0). This was then followed by a pH phosphate buffer wash (100 mM sodium phosphate, monobasic, 10 mM Tris and 0.05% triton X-100, pH 6.3). The protein was then eluted from the column with a pH buffer of 250 mM imidazole (250 mM imidazole, 100 mM sodium phosphate, monobasic, 10 mM Tris and 0.05% triton X-100, pH 5.9).

Final Formulation BASB034 was formulated through dialysis during the night with three 0.1% changes of triphone X-100 and 1x PBS, pH 7.4, to remove residual Gu-HCl and imidazole. The purified prolein was characterized and used to produce anibodies as described below.

SDS-PAGE Biochemical Characterizations and Western Stain Analysis The recombinant purified protein was resolved in 4-20% polyacrylamide gels and transferred electrophoretically to PVDF membranes at 100 V for 1 hour as previously described (Thebaine et al., 1979, Proc. Nail, Acad. Sci. USA 76: 4350-4354). The PVDF membranes after pretreated with 25 ml of saline regulated in its pH with Dulbecco's phosphate containing 5% dry milk without fat. All subsequent incubations were performed using this pre-traction pH regulator. The PVDF membranes were incubated with 25 ml of a 1: 500 dilution of preimmune serum or immune serum of rabbit ani-His for 1 hour at temperalura ambienle. Then, the PVDF membranes were washed twice with wash buffer (20 mM tris pH buffer, pH 7.5, containing 150 mM sodium chloride and 0.05% Tween-20). The PVDF membranes were incubated with 25 ml of a 1: 5000 dilution of peroxidase-labeled goat anti-rabbit IgG (Jackson Immuno Research Laboratories, Wesf Grove, PA) for 30 minutes at room temperature. Then, the PVDF membranes were washed four times with the washing pH regulator, and were developed with 3-amino-9-ethylcarbazole and urea peroxide as supplied by Zymed (San Francisco, CA) for 10 minutes each. "The results of an SDS-PAGE (FIG. 4) show a protein of approximately 60 kDa that is reactive to the antibody ani-RGS (His) through Western blots (FIG. 5) of the SDS-PAGE.

Protein Sequencing Amino-terminal amino acid sequencing of the purified protein was performed to confirm the production of the correct recombinant protein using well-defined chemical protocols in a Hewlett-Packard sequencer model G1000A with a 1090LC model and a Hewlett-Packard sequencer. Model 241 with an 1100LC model.

Example 5 Production of Antisera for recombinant BASB034 Polyvalent antisera directed against the protein BASB034 were generated through the vaccination of two rabbits with the purified recombinant BASB034 protein. Each animal was given a 3 immunization schedule intramuscularly (i.m.) of approximately 20μg of BASB034 proiein by injection (starting with Freund's adjunct auxiliary and followed by Freund's incomplete adjuvant) at approximately 21-day primers. The animals were bled before the first immunization ("pre-bled") on days 35 and 57. Anti-BASB034 protein tifulations were measured by ELISA using a purified recombinant BASB034 (0.5μg / well). The titration was defined as the highest dilution equal to or greater than 0.1 as calculated with the following equation: OD average of the two test sample of anisomer -OD average of the two test samples of the pH regulator. The smears after fresh immunizations were around 1000000. The antisera were used as the first antibody to identify the protein in a Western stain as described in example 4 above. Western staining shows the presence of the anti-BASB034 antibody in the sera of immunized animals (Figure 6).

Example 6 Analysis of the Flanking Regions that are not Coding of the BASB034 gene, and its exploitation for the expression of the modulated BASB034 gene The flanking regions that are not coding for the BASB034 gene contain important regulatory elements in the expression of the gene. This regulation takes place both at the transcriptional and translation levels. The sequence of these regions, either upstream or downstream of the open reading frame of the gene, can be obtained through DNA sequencing. This sequence information allows the determination of polar regulatory motifs such as the different promoter elements, terminator sequences, sequence elements capable of induction, repressors, elements responsible for phase variation, the shine-dalgarno sequence, regions with potential secondary structure involved in regulation, as well as other types of mofivos or regulatory sequences. This sequence information allows the modulation of the natural expression of the BASB034 gene. The upregulation of gene expression can be achieved by altering the promoter, the shine-dalgarno sequence, repressor elements or potential operators, or any other element involved. Also, the sub-regulation of expression can be achieved through similar types of modifications. Alternately, by changing the phase variation sequences, the expression of the gene can be placed under the confrol of phase variation, or it can be decoupled from this regulation. In another aspect, the expression of the gene can be placed under the control of one or more elements capable of induction by permifying the regulated expression. Examples of such regulation include, but are not limited to, induction through temperalure displacement, addition of base substrates such as selected carbohydrates or their derivatives, trace elements, vitamins, co-factors, metal ions, etc. Said modifications, as described above, can be introduced through different means. Modification of sequences involved in gene expression can be performed in vivo through random mutagenesis followed by selection for the desired phenotype. Another aspect is to isolate the region of interest and modify it through random mutagenesis, or site-directed mutagenesis, insertion or deletion muiagenesis. The modified region can then be reintroduced into the bacterial genome through homologous recombination, and the effect on gene expression can be determined. In other respects, the sequence knowledge of the infertile region can be used to replace or eliminate strains or from the nalural regulatory sequences. In this case, the target regulatory region is isolated and modified in order to contain the regulatory elements of another gene, a combination of regulatory elements of different genes, a synthetic regulatory region or any other regulatory region, or to eliminate selected parts of the regulatory sequences of wild type. These modified sequences can then be reintroduced into the bacterium through homologous recombination in the genome. A non-exhaustive list of preferred promoters that can be used for the upregulation of gene expression includes the PorA promoter, porB, IbpB, tbpB, p110, Ist, hpuAB of N. meningitidis or N. gonorroheae. In one example, the expression of the gene can be modulated by exchanging its promoter with a stronger promoter (through the isolation of the sequence upstream of the gene, in vitro modification of this sequence, and reintroduction into the genome through homologous recombination) . Overregulated expression can be obtained both in the bacterium as well as in other membrane vesicles that originate from (or are made of) the bacterium. In other examples, the aspects described can be used to generate recombinant bacterial strains with improved characteristics for vaccine applications. These may be, but are not limited to, attenuated strains, strains with high expression of selected antigens, strains with shocks (or reduced expression) of genes that interact with the immune response, strains with modulated expression of immunodominant proteins, strains with modulated diffusion of outer membrane vesicles. A direct region upstream of the BASB034 gene is given in the sequence of SEQ ID NO: 13. This sequence is one more aspect of the invention.

SEQUENCE INFORMATION Sequences of polynucleotide and polypeptide BASB034 SEQ ID NO: 1 BASB034 polynucleotide sequence of Moraxella catarrhalis of strain ATCC 43617 ATGAA ^ TtTCACTGtCTACATtGACTtTAtCTATTl ^ ACCAAATCCTGTGG < "-TTGT_GACG;.? AGTACGCAGT AAAGTGCGACACAATCAGCGTCTACTGATACGGCTAATCCTTTAGACGAACATGAACCAGAGCTtTATACGACAGCTTTA GAAAATAAAACCAT-CTOATTAACTGCTCAGCACTT ^^ TGGTGAGACGCC» «XK_T- TTAAAACCA ^ ^ CCCAGGTTATCtATC-AGAAACGACAGATCCGATTTTTTTAA AAACAGCrtGAATATGCAGCCAAACÍ TTTACACCACTGAGCT ATCAT_tGA_TrAG GTOVTC CGACCACACAATCCGA_OTATOTA GTCATGAAGCAAAACAATTTACCCCMATGAA__ CGTsCtCC8 ^ ^ ^ GA_ttAtc -TttsGATATACAC GCTGAs_ATtrAtsGGGGACGGAtt ct ^ sa .tcsc AAACTCT8TCCTCTTAGAGTACATGACTACCAGCCAGAGATTre ATGGa????? -XAGtCCGCAT-ATTGGC? TCGGTGCGGTACATCATTC CGTGC_TAtttGATGsCAGGCATGsAATGGAAAAACCTGACIX_TC ^? ^ TAGTGGC_AGCC TsCCACGCAT C_AGATGAT.UVrCCrGAtATCTTGGACTATTATO ^ ^ AAAATAAAAGTAAtATTTa_3GTACGGTA03CTATAATC? CSCT CCX3CTTCK_TAAGGsAATrA ^? tGsCTAtTtTCAAATATTTC? AGG GAC_ GCrrrGGCsTCGGACTTATG ^ p, AACX_ACtGGATGGGtCTATAA SEQ ID NO: 2 Polynucleotide sequence BASB034 of Moraxella catarrhalis deduced from the polynucleotide of SEQ ID NO: 1 M VS ST T SILSCFAIIAIQQAQAVPNPVAFVDEVRSEND GQDNELPIDVQ? ATQSASTDTANPLDEHEPELYTTAL ENKT LINCSALNQDI > 1RIACYDTLVHGETPAVIKtKRSIRLDEtiWQTIKGKPQVIYaETTDPIFLMGNEKG-1LTKKDA KQLEYAAKQFTPLS SFDLDRN TPLWSSRPHNP YVLPIFHHGKPNRSPNtPSHEAKQFTPNEFRAPELKFQVSV VKA AEDJ_WGtDSD WFGYTQQSH QIFNGKNSRPFRVHDYQPEIFI_TQPVYSDLPWDGKVRMIGMsAVHHSNGESAK SRSWN RAYLMAGME KNLTVMPRt GRIFKEGSsSQPDDNPDILDYYGYGDVRFLYQt, ENKSNISGTVRYNPRSGKGALQLDYVY P GKGISGYFQIFQsYGQSl, IDYNHEATSFGVG M NDW-TGL SEQ ID NO: 3 polynucleotide sequence of BASB034 of Moraxella catarrhalis strain Mc2908 of AtGAAAGTtTCACtGTCTACATtGACTTrATCTATTtTGCCAtGTtTTGCTAtCCtAGCCAtTCAsCAAGCACAAGCtGT ACAU ^ TCC_O.ssCA_T? _rTCA8AAG.ACsCM CGTC.ACTsA.ACGGCTAAtCCm AAAG.OCGACACAAtC ^ ^ GAAAATAAAACC? TGCrGATTAACTGCTCftGCACTTAATCAAGATATCATsa3TTTssCsTGCTATsAC? CTTTGsT TGGTGAGACGCC ^? ^ SCGGTAATTAAAACCAAGCGTTCCATTCGCCTTGATsAAACAATTT_sCAGACCATCAAA CCCAGGTTGTCTATC_vAGAAACGAC »sATCCGA_Tr.TrtAATGGG-A AAACAGCTtGAATATGCAGCCAAACAGTTrACACCACTGAGCTTATC ^^ ^ ^ GTCATCACGACCACACAATCsjATsTATGTATTGCCCATATTTATGCACGsTAAsCCtAATCGAAGCCCAAATACGCCCA GTCATGAAGCAAGACAATT ACCCCAAATGAATnTOtsCCCCT3AArrAAAATtTCAAGT_ GCTGAGGAT_TATGGGGsACGGATTCAGAtTTATGsTt GGGTATACACAGCAATCGCACtGsCAGATTtTTAATGGAAA AAACTCTCOTCCTTTTAGAGTACATGAT-AC CCAGAGATTGTCTTAACT ATGGCAAAGTCCGCATGATTsGCATGGOTGCGGTACATCATTCC ^? ^ A ^ CGTGC_TATTtsATGGCAGGCATGGAA_ssAAAAACC_t_ACrG_ TAGTGGCAGCC? GCCAGATGACAATCCTGATATCTTsGACTATtATsGTTATGGTsAtGTGCGTTttTTATATCAA-TAG AAAATAAAAGTAAtAT_tCAGGTACsGTACGCTAtAATCCACGCTCAGGCAAAGGTGCGTTsCAACTtsACTATGTCTAT CCGCTtGGTAAGGGAATTAGTGGCTATTTTC ATATT ?? TCAAsGCTATGGGCAGTCTTTGATTGAttATAAtCATsAGGC GAa_ \ GCTTTGGCGTCGGACTTATGCTTAACGACTGGATGGGTCTATAA SEQ ID NO: 4 BASB034 polynucleotide sequence of Moraxella catarrhalis deduced from the polynucleotide of SEQ ID NO: 3 MKVSLSTT _: _ IF PCFA_U "Q_AQA-PNPVAFVDHRVRSKND; QD_RE IN RM _NCSAI QDIMRI_ACYDTLVHGETPAVIKT.3SIR_JD_N:? _ ^^ KQLEYAA QFTP.SLSFDI RNGGGPLWSSRPHNPMYVLPIF HGKPNRSPNTPSHEARQFTPNEFRAPE KFQVSVÍVKA AEDLWGTDSDLWFGYTQQSHWQIFNGKNSRPFRVHDYQPEIFLTQPVYSDLPWDGKVRMIGMGAVHHSNGESAKLSRSWN RAYL AGME KNLTVHPRIWGRIFKEGSGSQPDDNPDILDYYSYGDVTÍF YQ ENKSNISGTVRYNPRSGKGA QLDYVY PLGKGIssYFQIFQGYsQSLIDYNHEAtSFGVGLM ND MGL SEQ ID NO: 5 Sequence polinucleóíido BASB034 from Moraxella catarrhalis strain Mc2913 the ATCAAAGt-TCACTGTCTAC TTsACTTTATCTATTTra ^ -_ ACOUUVrCCreTCK¡C TaTTGA8AAGTACGC AAAGTGCGAC ^ C ^ AtCAGC_GtcTAC-aA_ACsscTAATCCrrTAGACGAACATGAACCAGAGCt_ GAAAATAAAACCATGCTsATTAACTGC_T GCACTTAATavAsATATCATGCsTTtGGCGtGCTATGA_ACT_ T8TGAGACGCC ^ / ^ CGGTAATTAAAACCAAGCOTTCCAttCGCCT CCCAGGtTGTCTAtCAAGAAACGACAGAtCCGATtTtTTTAATGGGTAATGAAAAAGGCATGCrrGACCAAAAAAGATsCC AAACAGC_ GAATATGaVGCCAAACAGTrTA_?????. \ CC ^ CTsAGCritATCATTTGATTTAsACCGAAATAAtAC GTCATCACGACCACACAAtCCGATGTATGTATTGCCCATATTTATGCACGGTAAGCCTAATCGAAGCCCAAATACGCCCA GTCAtGAAGCAAGACAAtTtACCCCAAATX;? AATTTCX-TGCCCCT-AATTAAAATTtCAAGTTTCTGtTAAGGTTA GCTGAGGATTTATGGGGGACGGATtCAGATTTAtGGTTrGGATATACACAGC_ATCG < _ArTGGC GATTTTTAAtGGAAA AAACTC.TCG_CC_Trr AaAGTACATOATTACCAsCCAGAGAT_ ^ ATsGCAAAGTC8C TGATTGGCATGGGTscssTACATCAtTCa TGGTGAAAGTGCCAAACtGTCtcs s-TGCTTATTtGATGGC GGCATGGAAtGGAAAAACCTGACrroTCA_aCCA TAGTCGO _CATTTGGGGGraTATC ^ ^ ^ GCCAGCCAGATGACAATCCrrGATATC.TTCGAC ^^ AAAATAAAAGTAAtATpCAGGTACGGTACGCTATAATCCACGCT GGCAAA ccGCt GGtAAsGsAA_t- 5tssctA_tp????? CAAATAtttc_AGGCtA_GsscAGtctttGATtsAttAt GAc_AGcrrtGscGtcGGActtAtscxtAAcsActGGATGsGtctATAA SEQ ID NO: 6 Polynucleotide sequence BASB034 of Moraxella catarrhalis deduced from the polynucleotide of SEQ ID NO: 5 MKVSLSTLTLSILSCFAILAIQQAKAVPNPVAFVDEVRSENDLGQDNELPrDVQSATQSASTDTANPLDEH? PELYTTA ENKrrM INCSAIxNQDIMRIACYDtLVHGETPAVIKp-KRSIRLDETIW KQUS_AAKQFTPI L_FDLDRNNTPLWSSRPHNPtf .PIFMHGKPraSPNTPSH ^ ^ AEDLWGTDSDL FGYTQQSHMQIFNGKNSRPFRVHDYQPEIFLTQPVYSD PWDGKVRMIGMGAVHHSNGESAKLSRSWN RAYLMAsMEWKNLTVMPRIWGRIFKEGSGSQPDDNPDILDYYGYsDVRFLYQriENKSNISGTVRYNPRSG GALQLDYVY PLGKGISGYFQIFQGYGQS IDYNHEATSFGVGtM NDWMGL SEQ ID NO: 7 Polynucleotide sequence BASB034 of Moraxella catarrhalis of strain Mc2969 ATGAAAGT-TCACTGtCTACATTGACttTAtCTATTrc ACCAAATCCTGTGGCy ^^ "^ AAAGTGCGACACAATCGGCGTCTACTGATACGGCTAATCCTrtAGACGAACATGAACCAGAsCtTTATACGAC_AGCTTrA GAAAATAAAACC TCtTGACGAAGTACGCAGTsAAAATsATCT TGCTGAtTAACTGCTCAGC CTTAATCAAGATATCAtGCGtTTsGCGTsCTAtGACACTTTGsTGCA TGGTGAGACGCCAGCGGtAATTAAAACC AGCGTtCC TTCsCCTTGATsAAACAATTTGGCAsACCAtCAAAGsCAAAC CCCAGGTTGTCTATCAAGAAA ^ ^ GTCATCACGACCACAaVATC CAGATCCGATTTTTtTAATGGGTAATGAAAAAGGCAtGCrrGACC_AAAAAAGATGCC AAACAGCTraAATATGCAGCCAAACAGrrTACACC CTGAsCTTA_ SATGTATGTAtTGCCCAtATTTATGCACGGTAAGCCTAATCGAAGCCCAAAT GTCATGAAGCAAAAa ^^ ^ ^ GCTGAssATtTAtsssGsACGGATTCAGATTTATsGTTTsaATATACA <TTTACCCCAAAtGMTCTCGTCCTCCCsAGCTAAAATTTCAGGTTTCTGr?????; ^ __ ^ AAACTCTCGTCCTt TAGAGtAC tsACTACCAGCCAGAGATtTTCTTAACTCAACCTsTATACtCAGACTTACCATsGG ATGGCAAAGTCCsCATGATTGGC t.GGTGCsGtACATCATTCCAATsGTGAAAGTGCCAAACTGTCtCGCTCATGGAAT CsTsCTTATTtGATGGCAGGCAtGGAATsGAAAAACCTGACTGTCAtsCCACGCAT'rTGGsGsCGTAtCT-rTAAAGAGGG TAGTGGCAGCCAGCC ^ GATGATAAtCCTGATAtCTTXJGACrATTAtsGtTATGGTGATGtGCGTTTTTrATATCAACTAG AAAATAAAAGtAATATTTCAGsTACGsTACGCTATAAtCCACGCTCAGGa ?? \ GGTGCGtTGCAACTTGACTAtsTCtAT CCGCTraGTAAGsGAATTAGTGGCn:? ATTrTC AATATTtCAAGsCTATGGGCAGTCTTtsATtGAtTATAATCATGAGGC GA ^ VGCTTtGGCGTCGsAC_tATGCtTAACsACTGGAtGGGTCTATAA? SEQ ID NO: 8 Polynucleotide sequence BASB034 of Moraxella catarrhalis deduced from the polynucleotide of SEQ ID NO: 7 MKVSLST T SILPCFAILAIQQAQAVPNPVAFVDEVRSENDLGQDNEIiPIDVQSATQSASTDTANPLDEHEPELYTTAL ENKTaiINCSAI? Qdim Iacy? T hget AVIKTKR5IR_DETI QTIKG PQVV QETTDPIFLHs EKsMLT KDA QLEYAAKQFTPLSLSFDLDRNNTPLWSSRPHNP YVLPIFMHGKP RSPNTPSHEAKQFTPNEFRAPELKFQVSVKVKA AEDLWGTDSD WFGYTQQSH QIFNGKNSRPFRVHDYQPEIF TQPVYSDLP DsKVR IG-IGAVHHSNsESAKLSRS N RA GMEWK LtVM IWGRIFKEGSGSQPDDt GD ^ PDILD G RFL Q_, ENKSNISGtVR SGKGALaIJD NP V PLGKGISGYFQIFQGYGQS IDYNHEATSFGVGLMLNDWMGL SEQ ID NO: 9 GAT TTA AGA GTA TGT TAT GAT G SEQ ID NO: 10 GTA TGG GTT GAT CAA ATA CAG SEQ ID NO: 11 AAG GGC CCA ATT ACG CAG AGG GGA TCC CAAC GCT GTA CCA AAT CCT GTG GCA TTT GTT G SEQ ID NO: 12 AAG GGC CCA ATT ACG CAG AGG GTC GTA TTA TTA TAG ACC CAT CCA GTC GTT AAG CAT AAG SEQ ID NO: 13 ACTTssCGAAAATACCATTTATATCGATTGTGATGTTATAC sGCAGAt CTGCXMTGGCACtTATTGATGCTTTAGAACACXrGCAGCGTCGTAAAAAGCTTACCCAAGATCCGCTTT ^ ^ GCAGCGGTTTCn > 3TGGG_GTCAA-CA_U3GCCGTC ^ A ^ TATGCtAAtOSttsAAGATAAtsGCGTGATCATCACAttAAATsGACAAGTAAAAsACCCATTATTtTGGtGGTCGATGA ttTAAATG_GG_CATOACGCAGsCAGGTGsG_tTATTGAGATTCAAGG AAGCTAATscsAtGCTTGATTTGGCAsAGCtGGGAATTsGsCAGATTATCGAAGCCCAAAAGC TAtTGCTGCrGCTGGGtGtCTtGGtGGCAATCATTTX.TTTGATrGCACCCGTrrtttATGCAATCGGTGCGTT-sctttA _TrcCAGrrcTGGTAT_TG.GTTTAATAtTCAAAGGCAAAAAGC GATTACGTCCAAACTCT__OMA_TCATAACAAAtCACTAAC AAATGAC TTGTTGATCGGGGCATtsAATATCATTTTA ^ < .AGGTT ^ GTACTTTTGGGAAAGTC ^ TCAAMCI AATGCGsTsGCGGt ^ A_OGtcttTAACTCCACCCACCTAAC'i'Tll'TCl'TTG_ tsGATTtAAGAGTATGTTATGAtGGGCAGGAtt_TAtttTAA GTCATCArrTAAtGCAATCAGtTsTCCAGAGTAGCCGTTC Deposited materials A deposit containing a Moraxella catarrhalis strain Catlin has been deposited with the Culture Collection American Type American Type Culíure Colleclion (hereinafter adelanfe "ATCC") on June 21, 1997 and assigned the number of depósilo 43617 . the depósiío was Branhamella catarrhalis as descrifo (Frosch and Kolle) and is a collection of inserlo 15.-9.2 kb freeze drying parfir conslruida to isolate M. catarrhalis obtained from a transtracheal aspiration of a mining carbon with chronic bronchiias. The deposit is described in Animicrob. Agenís Chemother, 21: 506-508 (1982). The deposition of the Moraxella catarrhalis strain is referred to in the present as "the deposited strain" or as "the DNA of the deposited strain". The deposited strain contains a full-length BASB034 gene. A reservoir of the pMC-PLA1 vector consisting of the DNA of Moraxella catarrhalis in pQE30 has been deposited in the Collection of American Type Culíivo American Type Culture Collection (ATCC) on February 12, 1999 and assigned the deposit number 207099. The sequence of the polynucleotides contained in the deposited strain, or in the deposited clone, as well as the amino acid sequence of any polypeptide encoded by it , are being controlled in the case of any conflict with any description of sequences in the present. Deposits of the deposited strain-clone have been made under the terms of the Budapest Tray in the International Recognition of the Deposit of Microorganisms for the Purpose of Patent Procedures. The strains deposited will be irrevocably and without restriction or condition released to the public after the issuance of a patent. The deposited strains are provided merely as a convenience to those skilled in the art and not as an admission that a deposit is required for habilitation, as required in accordance with 35 U.S.C. § 112 LIST OF SEQUENCES < 110 > Smi.thKli.ne Beecham Biolsgicals S.A. < 120 > Novel Compounds < 130 > BM4S332 < 1S0 > 13 < 170 > FastSEQ for Windows Version 3.0 < 210 > 1 < 211 > 1329 < 212 > DNA < 213 > Mocaxella caearrhalis < 400 > 1 aegaaagtte cactgtctac atttgasttta tctattttgt catgttctgc tatcctagcc 60 attcagcaag cacaagctgt accaaatcct gtggcatttg ttgacgaagt acgcagtgaa 120 aattgatcttg ggcaagacaa tgaattaccc attgatgtcc aaagtgcgac acaatcagcg 180 Cctaccgata cggctaatco tttagacgaa caCgaaccag agctttatac gacagcttta 240 gaaaátaaaa ccatgctgat taactgctca gcacttaatc aagatatcat gcgtttggcg 300 ctttggtgca cgctatgaca tggtgagacg ccagcggtaa tcaaaaccaa gcgttccatc 3S0 cgectttgatg aaacaatetg gcagaccatc aaaggcaaac cccaggctac ctatcaagaa • 420 acgacagatc cgatttttttt aaCgggtaat gaaaaaggca cgccgaccaa aaaagatgcc 430 aaacagcttg aatatgcagc caaacagttt acaccactga gcteattcatt tgatttagac S40 cgaaataaca caccactctg gtcatcacga ccacacaatc cgatgtatgt attgcccata SOO tttatgcacg tcgaagccca gtaagcctaa aatacgccca gtcacgaagc aaaacaattt that accccaaatg aatctcgtgc tcccgagcta aaatctcagg tttctgctaa GGT aaagcc 720 gctgaggatt tatgggggac ggactcagac ttatggtttg gacacacaca gcaatcgcac 780 tggcagattt tcaatggaaa aaactctcgt ccttttagag tacacgacta ccagccagag 840 attctccta to ctcaacctgt atactcagac ttaccatggg acggcaaagt ccgcatgacc 900 ggcatgggcg cggtacatca tttccaatggt gaaagtgcca aactgccccg ctcatggaac 960 cgcgcttatc cgatggcagg cacggaacgg aaaaacctga ccgccacgcc acgcatctgg 1020 gggcgtatct tcaaagaggg tagtggcagc cagccagacg acaatcctga tatcttggac 1080 cactacggee acggtgaegc gcgceeetca tatcaactag aaaataaaag caacacccca 1140 ggtacggtac gcuataatcc acgctcaggc aaaggtgcgt tgcaacttga ctatgtctat 1200 ccgcttggta agggaattag tggct ttttt caaatatttc aaggctatgg gcagtcttcg 1260 actgattata atcatgaggc gacaagcttt ggcgtcggac ttatgcttaa cgactggatg 1320 ggcctataa 1329 < 210 > 2 < 211 > 442 < 212 > PRT < 213 > Moraxella catarrhalis < 400 > 2 Met Lys Val Ser Leu Ser Thr Leu Thr Leu Ser He Leu Ser Cys Phe 1 5 10 15 Wing He Leu Wing He Gln Gln Wing Gln Wing Val Pro Asp. Pro Val Wing 20 25 30 Phe Val Asp Glu Val Arg Ser Glu Asn Asp Leu Gly Gln Asp Asn Glu 3S 40 4S Leu Pro He Asp Val Gln Ser Wing Thr Gln Ser Wing Ser Thr Asp Thr 50 SS SO Wing Asn Pro Leu Asp Glu His Glu Pro Glu Leu Tyr Thr Thr Ala Leu 65 70 75 80 Glu Asn Lys Thr Met Leu He Asn Cys Ser Ala Leu Asn sln Asp He 85 90 95 Met Arg Leu Ala Cys Tyr Asp Thr Leu Val His Gly Glu Thr Pro Ala 100 IOS 110 Val He Lys Thr Lys Arg Ser He Arg Leu Asp Glu Thr He Trp G n H5 120 125 Thr He Lys Gly Lys Pro Gln Val He Tyr Gln Glu Thr Thr Asp Pro 13 ° 135 140 He Phe Leu Met Gly Asn GIu Lys Gly Met Leu Thr Lys Lys Asp Wing 145 1S0 1S5 160 Lys Gln Leu Glu Tyr Wing Wing Lys Gln Phe Thr Pro Leu Ser Leu Ser 1S5 170 175 Phe Asp Leu Asp Arg Asn Aan Thr Pro Leu Trp Ser Ser Arg Pro His 180 185 190 Asn Pro Met Tyr Val Leu Pro He Phe Mee His Gly Lys Pro Asn Arg 195 200 205 Ser Pro Asn Thr Pro Ser His Glu Wing Lys Gln Phe Thr Pro Asn Glu 210 21S 220 Phe Arg Wing Pro Glu Leu Lys Phe sln Val Ser Val Lys Val Lys Wing 22S 230 23S 240 Wing Glu Asp Leu Trp Gly Thr Asp Ser Asp Leu Trp Phe Gly Tyr Thr 245 250 255 Glp Gln Ser His Trp Gln He Phe Asn Gly Lys Asn Ser Arg Pro Phe 260 26S 270 Arg Val His Asp Tyr Gln Pro Glu He Phe Leu Thr Gln Pro Val Tyr 275 280 285 Ser Asp Leu Pro Trp Asp Gly Lys Val Arg Met He Gly Met Gly Wing 290 29S 300 Val His His Ser Asn Gly Glu Ser Wing Lys Leu Ser Arg Ser Trp Asn 305 310 315 320 Arg Wing Tyr Leu Met Wing Gly Mett Glu Trp Lys Asn Leu Thr Val Met 325 330 335 Pro Arg He Trp Gly Arg He Phe Lys Glu Gly Ser Gly Ser Gln Pro 340 345 350 Aßp Asp Asn Pro Asp He Leu Asp Tyr Tyr Gly Tyr Gly Asp Val Arg 355 360 365 Phe Leu Tyr Gln Leu Glu Asn Lys Ser Asn He Ser sly Thr Val Arg 370 375 380 Tyr Asn Pro Arg Ser Gly Lyß Gly Ala Leu Gln Leu Asp Tyr Val Tyr 385 390 395 400 Pro Leu Gly Lys Gly He Ser Gly Tyr Phe Glp He Phe Gln Gly Tyr 405 410 41S Gly Gln Ser Leu He Asp Tyr Asn His Glu Wing Thr Ser Phe Gly Val 420 425 430 Gly Leu Mett Leu Asn Asp Trp Mett Gly Leu 43S 440 < 210 > 3 < 211 > 1329 < 212 > DNA < 213 > Moraxella caear.fialis < 40Q > 3 atgaaagttc cacttgcccac accgacccta cctactttgc catgctttgc tatcctagcc 60 actcagcaag cacaagccgc accaaacccc gcggcatccg tttgacgaagt acgcagcaaa 120 aatgatcttg ggcaagacaa ttgaattactc attggtgtac aaagtgcgac acaatcagcg 180 cccaccgaca cggctaaCcc tctagacgaa cacgaaccag agccccacac gacagcccca 240 gaaaacaaaa ccatgctgatt taactgcttca gcactttaatc aagattatcat gcgttttggcg 300 ctttggtgca tgctatgaca tggtgagacg ccagcggtaa ttaaaaccaa gcgttccatt 360 cgccccgattg aaacaatttg gcagaccacc aaaggcaaac cccaggtcgc ctatcaagaa 420 acgacagatc cgattttttt aatgggtaat gaaaaaggsa tgctgaccaa aaaagatgcc 430 aaacagcttg aatatgcagc caaacagctc acaccactga gcttattcatt tgattttttagac 540 cgaaataata caccgctctg gttcatcacga ccacacaattc cgatgttatgc atttgcccata 600 gtaagcctaa tccaegcacg tcgaagccca aatacgccca gttcattgaagc aagacaactt 660 accccaaatg aattttcgtgc ccccgaattta aaattttcaag ttttcttgttaa ggttaaagct 720 gctgaggatt tacgggggac ggatttcagat ttatggcttg ggtatacaca gcaatcgcac 780 tggcagattt tttaatggaaa aaactctcgt ccttttagag tacatgattta ccagcc agag 840 attetcttaa ctcaacccgt gtacccagac ttaccacggg acggcaaagt ccgcatgact 900 cggtacatca ggcatgggtg tttccaatggt gaaagtgcca aaccgtcttcg cccatggaat 960 cgtgctcattt tgatggcagg cacggaatgg aaaaacctga cttgtcatgcc acgcacccgg 1020 gggcgttatCC ttaaagaggg tagttggcagc cagccagacg acaatcctga tattcttttggac 1080 catttatggttt atggtgatgt gcgttttttta ttatcaacttag aaaattaaaag taattacttca 1140 ggcacggcac gccaeaattcc acgctcaggc aaaggtgcgt ttgcaactcga ccattgccttac 1200 ccgctttggta agggaatttag tggstatttt caaatatttc aaggctatgg gcagtctttg 1260 accgatcaca attcatgaggc gacaagcttc ggcgtcggac tcattgcctaa cgaccggacg 1320 ggtcttacaa 1329 < 210 > 4 < 211 > 442 < 212 > PRT < 213 > Mtoraxella catarrhalis < 400 > 4 Met Lys Val Ser Leu Ser Thr Leu Thr Leu Ser He Leu Pro Cys Phe 1 5 10 15 Ala He Leu Ala lie Gln Gln Ala Gln Ala Val Pro Aßn Pro Val Ala 20 25 30 Phe Val Asp Glu Val Arg Ser Ly3 A3n Asp Leu Gly Gln Asp Asn Glu 35 40 45 Leu Leu He Gly Val Gln Ser Wing Thr Gln Ser Wing Ser Thr Asp Thr SO SS 60 Wing Asn Pro Leu Asp Glu His Glu Pro Glu Leu Tyr Thr Thr Ala Leu 65 70 5 ao Glu Asn Lys Thr Met Leu He Asn Cys Ser Wing Leu Asn Gln Asp He 3S 90 95 Met Arg Leu Wing Cys Tyr Asp Thr Leu Val His Gly Glu Thr Pro Wing 100 IOS 110 Val He Lyß Thr Lyß Arg Ser He Arg Leu Asp Glu Thr He Trp Gln 115 120 125 Thr He Lys Gly Lys Pro Gln Val Val Tyr Gln Glu Thr Thr Asp Pro 130 135 140 He Phe Leu Met Gly Asn Glu Lys Gly Met Leu Thr Lys Lys Asp Ala 145 150 15S 160 Lys Gln Leu Glu Tyr Ala Wing Lys Gln Phe Thr Pro Leu Ser Leu Ser 165 170 175 Phe Asp Leu Asp Arg Asn Asn Thr Pro Leu Trp Ser Ser Arg Pro His 180 185 190 Asn Pro Met Tyr Val Leu Pro He Phe Met His Gly Lys Pro Asn Arg 195 200 205 Ser Pro Asn Thr Pro Ser His Glu Ala Arg Gln Phe Thr Pro Asp Glu 210 215 220 Phe Arg Wing Pro Glu Leu Lys Phe Gln Val Ser Val Val Lys Wing 225 230 235 240 Wing Glu Asp Leu Trp Gly Thr Asp Ser Asp Leu Trp Phe Gly Tyr Thr 245 250 255 Gln Gln Ser His Trp Gln He Phe Asn Gly Lys Asn Ser Arg Pro Phe 260 265 270 Arg Val His Asp Tyr Gln Pro Glu He Phe Leu Thr Gln Pro Val Tyr 275 280 285 Ser Asp Leu Pro Trp Asp Gly Lyß Val Arg Mett He Gly Met Gly Ala 290 295 300 Val His His Ser Asn Gly Glu Ser Ala Lys Leu Ser Arg Ser Trp Asn 305 310 315 320 Arg Ala Tyr Leu Met Ala Gly Met Glu Trp Lya Asn Leu Thr Val Met 325 330 33S Pro Arg He Trp Gly Arg He Phe Lys Glu Gly Ser Gly Ser Gln Pro 340 345 350 Asp Asp Asp Pro Asp He Leu Asp Tyr Tyr Gly Tyr Gly Asp Val Arg 355 360 365 Phe Leu Tyr Gln Leu Glu Asn Lys Ser Asn He Be Gly Thr Val Arg 370 375 380 Tyr Asn Pro Arg Ser Gly Lys Gly Ala Leu Gln Leu Asp Tyr Val Tyr 385 390 395 400 Pro Leu Gly Lys Gly He Ser Gly Tyr Phe Gln He Phe Gln Gly Tyr 405 410 415 Gly Gln Ser Leu He Asp Tyr Asn His Glu Wing Thr Ser Phe Gly Val 420 425 430 Gly Leu Met Leu Asn Asp Trp Mett Gly Leu 435 440 < 210 > 5 < 211 > 1329 < 212 > DNA < 213 > Moraxella ca arrhalis < 400 > 5 acgaaagttt cactgtctac attgactttta tctattttgt catgttttgc catcctagcc 60 attttcagcaag caaaagctgtt accaaatcct gtggcattttg ttgacgaagtt acgcagtgaa 120 aacgatccttg ggcaagacaa tgaatttaccc attcgatgtcc aaagttgcgac acaatcagcg 180 tctactgata cggctaatcc tctagacgaa cattgaaccag agctttatac gacagcttta 240 gaaaataaaa ccattgccgac taacttgctca gcacctaatc aagatatcat gcgttttggcg 300 tgctatgaca CTTT gttgca tggttgagacg ccagcggttaa ttaaaaccaa gcgctccattt 360 cgccttgatg aaacaattt gcagaccattc aaaggcaaac cccaggtttgt cttattcaagaa 420 acgacagatc cgattttttc aatgggtaac gaaaaaggca cgcCgaccaa aaaagacgcc 480 aaacagcttg aatatgcagc caaacagttt acaccactga gcttatcatt tgatttagac S40 cgaaataata caccactttg gccatcacga ccacacaatc cgattgcacgt atttgcccata 600 ttttt ttgcacg gtaagccttaa aatacgccca tcgaagccca gtcatgaagc aagacaactt 6S0 accccaaattg aatttttsgtgc ccctgaatta aaatttttcaag ggtttaaagstt tttctgtttaa 720 gccgaggattt tatgggggac ggatccagat tttacggtctg gatattacaca gcaatcgcac 780 tggsagatttt ccaattggaaa aaacttctcgt cctttttagag tac atgatta ccagccagag 840 attttttcttaa ctcaacctgt atactcagac ttaccatggg atggcaaagt ccgcatgatt 900 cggtacattca ggcatgggtg tttccaattggtt gaaagttgcca aaccgtcttcg cttcatggaac 960 cgcgcttatt tgatggcagg catggaatgg aaaaacctga ctgtcatgcc acgcatttgg 1020 gggcgcatct ttaaagaggg tagtggcagc cagccagattg acaaCcccga caccttggac 1080 tatttatggtt atggtgatgt gcgttttttta tatcaactag aaaattaaaag ttaacacccca 1140 ggtacggtac gcttataattcc acgctcaggc aaaggtgcgtt tgcaacttga ctatgtcttatt 1200 ccgctttggta agggaatttag tggccatttt caaatatttttc aaggctacgg gcagttctttg 1260 attgatttatta accattgaggc gacaagcctt ggcgtcggac tttattgcccaa cgacttggatg 1320 ggtctataa 1329 < : 210 > 6 < 211 > 442 < 212 > PRT < 213 > Moraxella caearrhalis, 400 > 6 Met Lys Val Ser Leu Ser Thr Leu Thr Leu Ser He Leu Ser Cys Phe 1 5 10 15 Ala He Leu Ala He Gln Gln Ala Lys Ala Val Pro Asn Pro Val Ala 25 30 Phe Val Asp Glu Val Arg Ser Glu Asn Asp Leu Gly Gln Asp Asn Glu 40 45 Leu Pro He A3p Val Gln Ser Ala Thr Gln Ser Ala Ser Thr Asp Thr SO SS 60 Wing Asn Pro Leu Asp slu His Glu Pro Glu Leu Tyr Thr Thr Ala Leu 6S 70 75 80 Glu Asn Lys Thr Met Leu He Asn Cys Ser Ala Leu Asn Gln Asp He 85 90 95 Met Arg Leu Ala Cyß Tyr Asp Thr Leu Val His Gly Glu Thr Pro Ala 100 IOS 110 Val He Lys Thr Lys Arg Ser He Arg Leu Asp Glu Thr He Trp Gln US 120 125 Thr He Lys Gly Lys Pro Gln Val Val Tyr Gln Glu Thr Thr Asp Pro 130 13S 140 He Phe Leu Met Gly Asp Glu Lys Gly Met Leu Thr Lys Lys Asp Ala 145 150 155 160 Lys Gln Leu Glu Tyr Ala Wing Lys sln Phe Thr Pro Leu Ser Leu Ser 165 170 175 Phe Asp Leu Aßp Arg Asn Asn Thr Pro Leu Trp Ser Ser Arg Pro His 180 18S 190 Aßn Pro Met Tyr Val Leu Pro He Phe Met His Gly Lys Pro Asn Arg 195 200 205 Ser Pro Asn Thr Pro Ser His slu Ala Arg Gln Phe Thr Pro Asn Glu 210 215 220 Phe Arg Wing Pro Glu Leu Lys Phe Gln Val Ser Val Val Lys Wing 225 230 235 240 Wing Glu Asp Leu Trp Gly Thr Asp Ser Asp Leu Trp Phe Gly Tyr Thr 24S 250 2SS Glp Gln Ser His Trp Gln He Phe Asn Gly Lys Asp Ser Arg Pro Phe 260 265 270 Arg Val His Asp Tyr Gln Pro Glu He Phe Leu Thr Gln Pro Val Tyr 27S 280 285 Ser Asp Leu Pro Trp Asp Gly Lys Val Arg Het He Gly Met Gly Wing 390 295 300 Val His His Ser Asn Gly Glu Be Wing Lys Leu Ser Arg Ser Trp Asn 305 310 315 320 Arg Wing Tyr Leu Mett Wing Gly Mett Glu Trp Lys Asn Leu thr Val Met 325 330 335 Pro Arg He Trp Oly Arg He Phe Lys Glu Gly Ser Gly Ser Gln Pro 340 345 350 Asp Asp Asn Pro Asp He Leu Asp Tyr Tyr Gly Tyr Gly Asp Val Arg 355 360 365 Phe Leu Tyr sln Leu Glu Asn Lys Ser Asn He Ser Gly Thr Val Arg 370 37S 380 Tyr Asn Pro Arg Ser Gly hya Gly Ala Leu Gln Leu Asp Tyr Val Tyr 385 390 39S 400 Pro Leu Gly Lys Gly He Ser Gly Tyr Phe sln He Phe sln Gly Tyr 40S 410 415 Gly Gln Ser Leu He Asp Tyr Asn His Glu Wing Thr Ser Phe Gly Val 420 425 430 Gly Leu Mee Leu Asn Asp Trp Met Gly Leu 435 440 < 210 > 7 < 211 > 1329 < 12 > DNA < 213 > Moraxella cacarrhalis < 400 >; 7 acgaaagttt cactgtccac attgacttt ttctatttttgc catgtttttttgc catccttagcc 60 attcagcaag cacaagctgt accaaattcct gtggcatttttg ttttgacgaagt acgcagttgaa 120 aatgatctttg ggcaagacaa tgaattaece attgatgtcc aaagttgcgac acaattcggcg 180 ttctactgata cggctaattcc tttaga gaa cattgaaccag agetttacae gacagcttttta 240 gaaaataaaa ccatgctgat ttaacttgcttca gcacccaatc aagacatcat gcgttcggcg 300 cctttggttgca tgctatgaca ttggcgagacg ccagcggtaa ttaaaaccaa gcgctccact 360 cgccttgatg aaacaattcg gcagaccattc aaaggcaaac cccaggttcgtt cttaccaagaa 420 acgacagatc cgatttttttttt aatgggtaat gaaaaaggca tgctgaccaa aaaagatgcc 480 aaacagcttg aacacgcagc caaacagttt acaccaccga gcctatcact CgatCCagac S40 cgaaataata caccaccttg gtcatcaega ccacacaatc cgattgcacgtt atttgcccata 600 gt tttacgcacg agcctaa aacacgccca tcgaagccca gtcatgaagc aaaacaaCCC 660 accccaaatg aacctcgcgc ccccgagcca aaaccccagg tccctgccaa ggttt aagcc 720 gctgaggatc tacgggggac ggactcagac ccacggctcg gacatacaca gcaattcgcac 780 tggcagattt ttaatggaaa aaactcttcgt ccttttagag tacattg minutes ccagccagag 840 actttcttaa ctcaacctgt atactcagac ttaccatggg atggcaaagt ccgcattgatttt 900 ggcatgggtg cggttacatca ttccaatggt gaaagttgcca aactgtctcg ctcatggaat 960 cgCgcttttatttt ttgattggsagg catggaatgg aaaaacctga ctgtcatgcc acgcatttgg 1020 gggcgtattct ttaaagaggg tagtggcagc cagccagatg ataatcctga tatctttggac 1080 tattatggttt atggttgatgt gcgtttttt tatcaactag aaaataaaag taatattctca 1140 ggtacggtac gctataaccc acgcttcaggc aaaggttgcgc tgcaactcga ctatgtctat 1200 ccgctttggta agggaattag tggctatttttt caaattatttttc aaggctatgg gcagttcettg 1260 atttgatttata attcatgaggc gacaagcttt ggcgtcggac ttatgcccaa cgactggacg 1320 ggtccataa 1329 < 210 > 8 < 211 > 442 < 212 > PRT < 213 > Moraxella catarrhalis < 4QQ > 8 Met Lys Val Ser Leu Ser Thr Leu Thr Leu Ser He Leu Pro Cys Phe 1 5 10 15 Wing He Leu Wing He Gln Gln Wing Gln Wing Val Pro Asn Pro Val Wing 20 25 30 Phe Val Asp slu Val Arg Ser Olu .Asn , Aßp Leu Gly Oln Asp Asn slu 35 40 45 Leu Pro He Asp Val Gln Ser Ala Thr Gln Ser Ala Ser Thr Asp Thr 50 S5 60 Wing Asn Pro Leu Asp Olu His Glu Pro Glu Leu Tyr Thr Thr Ala Leu 65 70 7S 80 Glu Asn Lys Thr Mett Leu He Asn Cys Ser Ala Leu Asn Gln Asp He 85. 90 95 Met Arg Leu Wing Cys Tyr Asp Thr Leu Val Hiß Gly slu Thr Pro Wing 100 105 110 Val He Lys Thr Lys Arg Ser He Arg Leu Asp Glu Thr He Trp Oln 115 120 125 Thr He Lys Oly Lys Pro ain Val Val Tyr sln Glu Thr Thr Asp Pro 130 135 140 He Phe Leu Met Gly Asn Glu Ly3 Gly Met Leu Thr Lys Lys Asp Ala 145 150 1S5 1S0 Lys Gln Leu Glu Tyr Ala Ala Lys Gln Phe Thr Pro Leu Ser Leu Ser 165 170 175 Phe Asp Leu Asp Arg Asn Asn Thr Pro Leu Trp Ser Ser Arg Pro His 180 185 190 Asn Pro Mett Tyr Val Leu Pro He Phß Mßt His Gly Lys Pro Asn Arg 195 200 205 Ser Pro Asn Thr Pro Ser His Glu Ala Lys Gln Phe Thr Pro Asn Olu 210 21S 220 Phe Arg Ala Pro Glu Leu Lys Phe Glp Val Ser Val Ly3 Val Lys Ala 225 230 235 240 Wing Glu Asp Leu Trp Gly Thr Asp Ser Asp Leu Trp Phe Oly Tyr Thr 24S 2S0 255 Oln sln Ser His Trp Oln He Phe Asn Oly Lys Asn Ser Arg Pro Phe 260 265 270 Arg Val His Asp Tyr Oln Pro Glu He Phe Leu Thr Gln Pro Val Tyr 275 280 28S Ser Aßp Leu Pro Trp Asp sly Lys Val Arg Mett He Gly Met Gly Ala 290 295 300 Val His His Ser Asn Gly slu Ser Ala Lys Leu Ser Arg Ser Trp Asn 305 310 315 320 Arg Ala Tyr Leu Met Ala Gly Met slu Trp Lys Asn Leu Thr Val Met 325 330 335 Pro Arg He Trp Oly Arg He Phe Lys Glu sly Ser Sly Be sln Pro 340 34S 3S0 Asp Asp Asp Pro Asp He Leu Asp Tyr Tyr Oly Tyr sly Asp Val Arg 355 360 365 Phe Leu Tyr Oln Leu slu Asn Lys Ser Asn He Ser Gly Thr Val Arg 370 375 380 Tyr Asn Pro Arg Ser Gly Lys Oly Ala Leu sln Leu Asp Tyr Val Tyr 385 390 395 400 Pro Leu sly Lys Gly He Ser Gly Tyr Phe Gln lie Phß sln sly Tyr 405 410 415 Oly Gln Ser Leu He Asp Tyr Asn His Glu Wing Thr Ser Phe Oly Val 420 425 430 Gly Leu Met Leu Asn Asp Trp Met Gly Leu 435 440 < 210 > 9 < 211 > 22 < 212 > DNA < 213 > Artificial Sequence < 220 > < 223 > Initiator sequence < 400 > 9 gatttttaagag eatgtcattga tg 22 < 210 > 10 < 211 > 21 < 212 > DNA < 213 > Artificial Sequence < 220 > < 223 > Initiator sequence < 400 > 10 gtattgggcttg attcaaataca g 21 < 210 > 11 < 211 > S8 < 212 > DNA < 213 > Artificial Sequence < 220 > < 223 > Oligonuoleotide < 400 > 11 aagggcccaa tttacgcagag gggatcccaa gctgtaccaa atcctgtggc atttgtttg 58 < 210 > 12 < 211 > 60 < 212 > DNA < 213 > Artificial Sequence < 220 > < 223 > Oligonucleotide < 400 > 12 aagggcccaa ccacgcagag ggtcgacCtta Cttacagaccc acccagccgc taagcacaag 60 < 210 > 13 < 211 > 1000 < 212 > DNA < 213 > Moraxßlla cacarrhalis < 400 > 13 actttggcgaa aattaccattt atatcgattg Cgatgttatta caggcagatg gcggttasaeg 60 cacagccagt atcagtggtg cttgcggtggc acttatttgat gcttttagaac actttgcagcg 120 ccgcaaaaag ctttacccaag attccgcctttt gggctttggttg gcagcggtttt ccgcgggtgt 180 taatcaaggc cgtgtattgc ttatgctgaa ttgatttgga gtgataccga gattcaacttt 240 ttttaaatgttg gtcattgacgc aggcaggtgg gttttattttgag atttcaaggca cagcagaaga 300 aaagccatttt acccgtgcttg aagctaatgc gatgcttgat ttggcagagc tgggaattttgg 360 gcagaecatc gaagcccaaa agcaagttatt aggcttggttga tatgcttaatc gttgaagata 420 atggcgtgatt catcacatta aattggacaag taaaagaccc atttatttttgg cggtcgacga 480 tattgcttgctt gcttgggtgtc ttggtggcaa tcatttgttt gattgcaccs gtttttttatg 540 caatcggttgc gttggcttta tttttgcagtttg ttggcatttgt gcttaatatt caaaggcaaa 600 aagccaaaac tttgtcattattg ttttcacaag gtcgcttgaa GAIT cgtcs aaacgctttg 660 caaatcacta agatttcataa accttttattcag catcggcaac aattattcegcc aaagattaaca 720 AAAT CCRA tgtttgatcgg ggcatt TAs attcatttttac aggtttttgct gatgaccgttg 780 aaattaatat agccaaacag gtacttt tgg gaaagtcaat caaaaccaat gcggtggcgg 840 taacatttggc taagtagttg tcgtgataca gacaggttgg atggtctttta actccaccca 900 cctaactttttt ttccttgttttg gatttttaagag tatgtcatga tgggcaggatt tttatttttttaa 960 gccatcatctt aatgcaattca gccgtccaga gtagccgttc 1000

Claims (26)

1. An isolated polypeptide comprising an amino acid sequence, which has at least 85% identity with the amino acid sequence selected from the group consisting of. SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6 and SEQ ID NO: 8.

2. An isolated polypeptide according to claim 1, wherein the amino acid sequence has at least 95% identity with the amino acid sequence selected from the group consisting of: SEQ ID NO: 2, SEQ ID NO: 4 , SEQ ID NO: 6 and SEQ ID NO: 8.

3. The polypeptide according to claim 1, comprising the amino acid sequence selected from the group consisting of: SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6 and SEQ ID NO: 8.

4. A polypeptide isolated from SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6 and SEQ ID NO: 8.

5. An immunogenic fragment of the polypeptide according to any of claims 1 to 4, wherein the immunogenic fragment is capable of producing an immune response (if necessary when coupled to the vehicle) which recognizes the polypeptide of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6 and SEQ ID NO: 8.

6. An isolated polynucleotide comprising a nucleotide sequence encoding a polypeptide having at least 85% identity to the amino acid sequence of SEQ ID NO: 2, 4, 6 or 8 over the entire length of SEQ ID NO. : 2, 4, 6 or 8 respectively; or a nucleotide sequence complementary to said isolated polynucleotide.

7. An isolated polynucleotide comprising a nucleotide sequence having at least 85% density with a nucleoide sequence encoding a polypeptide of SEQ ID NO: 2, 4, 6 or 8 over the entire coding region; or a nucleotide sequence complementary to the isolated polynucleotide.

8. An isolated polynucleotide comprising a nucleotide sequence having at least 85% identity to that of SEQ ID NO: 1, 3, 5 or 7 over the entire length of SEQ ID NO: 1, 3, 5 or 7, respectively; or a complementary nucleotide sequence for said isolated polynucleotide.

9. The isolated polynucleotide according to any of claims 6 to 8, wherein the identity is at least 95% for SEQ ID NO: 1, 3, 5 or 7.

10. An isolated polynucleotide comprising the sequence of nucleoid encoding the polypeptide of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6 or SEQ ID NO: 8.

11. An isolated polynucleotide comprising the polynucleotide of SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5 or SEQ ID NO: 7.

12. An isolated polynucleotide comprising an isolated nucleotide sequence encoding the polypeptide of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6 or SEQ ID NO: 8, which may be obtained by sorting an appropriate collection under severe hybridization conditions with a probe labeled having the sequence of SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5 or SEQ ID NO: 7 or a fragment thereof.

13. An expression vector comprising an isolated polynucleotide according to any of claims 6-12.

14. A recombinant living microorganism comprising an expression vector according to claim 13.

15. A host cell comprising the expression vector of claim 13.

16. A recombinant membrane of the microorganism of claim 14, expressing a polypeptide. isolated comprising an amino acid sequence having at least 85% identity with the amino acid sequence selected from the group consisting of: SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6 or SEQ ID NO : 8

17. A process for producing a polypeptide comprising an amino acid sequence that has at least 85% identity with the amino acid sequence selected from the group consisting of: SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6 or SEQ ID NO: 8, which comprises culturing a host cell of claim 15 under conditions sufficient for the production of said polypeptide and recovering the polypeptide from the culture medium.

18. A process for expressing a polynucleotide according to any of claims 6-12, which comprises transforming a host cell with the expression vector comprising at least one of the polynucleotides and culturing the host cell under conditions sufficient for the expression of any of the polynucleotides.

19. A vaccine composition comprising an effective amount of the polypeptide of any of claims 1 to 5 and a pharmaceutically acceptable carrier.

20. A vaccine composition comprising an effective amount of the polypeptide of any of claims 6 to 12 and a pharmaceutically acceptable carrier.

21. The vaccine composition according to any of claim 19 or 20, wherein said composition comprises at least one Moraxella catarrhalis antigen.

22. A nonspecific antibody to the polypeptide or immunological fragment according to any of claims 1 to 5.

23. A method for the diagnosis of a Moraxella infection, which comprises identifying a polypeptide according to any of claims 1- 5 or an antibody that is immunospecific for said polypeptide, present within a biological sample of an animal suspected of having said infection.

24. The use of a composition comprising an immunologically effective amount of a polypeptide according to any of claims 1-5, in the preparation of a medicament for use in generating an immune response in an animal.

25. The use of a composition comprising an immunologically effective amount of a polypeptide according to any of claims 6-12, in the preparation of a medicament for use in generating an immune response in an animal.

26. A therapeutic composition useful for the treatment of humans with Moraxella catarrhalis disease, comprising at least one antibody directed against the polypeptide of claims 1-5 and a suitable pharmaceutical vehicle.