MXPA97009473A - Gene of adherence of helicobacter pylori and polipeptide coded by me - Google Patents

Abstract

The present invention relates to an adhesion gene from Helicobacter pilory, a polypeptide encoded therein and antibodies against the polypeptide.

Description

GENE OF ADHERENCE OF Helicobacter Oylori AND POLYPEPTIDE CODED BY THE SAME DESCRIPTION OF THE INVENTION The present invention relates to a method for identifying secretory genes and in particular adhesion genes of Helicobacter pylori. In addition, the invention relates to a gene bank suitable for identifying H. pylori secreting genes, polynucleotides and polypeptides obtainable from this gene bank, in particular alpA-Helicobacter pylori gene and the polypeptide which it encodes. These polynucleotides and polypeptides can be used to diagnose, prevent and treat Helicobacter infection. The presentation of spiral bacteria in the human gastric mucous membrane has been known for a long time (Bizzozero, 1893). The fact that these are pathogenic germs, however, was not taken into account until the successful isolation and culture of this bacterium by Marshall and Warren (Warren and Marshall, 1983; Marshall et al., 1984) of the gastric mucous membrane of a patient with a gastric ulcer (ulcus ventriculi). In the first analysis we expect the isolated microorganisms to be gram-negative spiral materials with an extremely high motility REF: 26233 and an unusual ability to be susceptible to survive in a strongly acidic environment (up to ca. pH 1.5). The germs which originally were called Campylobacter pylori were originally classified based on their biochemical and morphological characteristics in the newly established genus "Helicobacter" (Goodwin et al., 1989). The importance of Helicobacter pylori infection and the implications of this discovery have already become clear in the following years. Epidemiological investigations by Taylor and Blaser (1991) have shown that H. pylori infection occurs worldwide and that ca. 50% of the population is infected with this bacterium. The infection rate is higher in developing countries than in industrialized countries. In addition, it has been observed that the probability of chronic H. pylori infection increases markedly with increasing age. Therefore, H. pylori infection is among the most frequent chronic bacterial infections in humans. It is now known that infection inevitably leads to the destruction of bacterial gastritis (type B gastritis) in humans. In addition, it is assumed that H. pylori also plays a causal role in the development of gastric and duodenal ulcers (ulcus ventriculi and ulcus duodeni) as well as some forms of gastric carcinoma (adenocarcinoma) (Lee et al., 1993; Solnick and Tompkins, 1993). Even the MALT lymphoid (lymphoid tissue associated with mucosa) of the stomach which occurs more rarely and is considered as a precursor of B-cell tumors of the immune system and also probably as a result of H. pylori infection. An antibacterial treatment of such patients with successful eradication (total elimination) of H. pylori leads to the healing of gastric ulcers as well as a low degree of MALT lympholas (Sipponen and Hyvárinen, 1993, Isaacson and Spencer, 1993, Stolte and Eidt, 1993 ). A sequel of a long-term infection with H. pylori is atrophic gastritis, a degeneration of the mucosa, of acid-producing cells or pepsin of the stomach epithelium which has been considered as a precancerous lesion. According to the statistics of the types of cancer which occur most frequently throughout the world in 1980, gastric carcinoma has the second place, with a tendency to decline (Parkin et al., 1988). Recently two studies have shown a statistically significant correlation between H. pylori infections and the presentation of gastric carcinoma (intestinal type); both came to the conclusion that ca. 60% of all gastric carcinomas that occur are probably due to H. pylori infection (Parsonnet et al., 1991; Nomura et al., 1991). In addition, research conducted by Sipponen (1992) shows that in many industrialized countries more than 20% of infected people get a stomach or duodenal ulcer during their life while the risk is negligibly small in people with normal gastric mucosa. This means that these frequent gastroduodenal diseases can be considered as infectious diseases and can be treated appropriately (Alper, 1993). A treatment which eliminates a chronic infection of H. pylori and is already present would lead to the healing of a gastritis, a gastric or duodenal ulcer or MALT lymphomas. Therefore, a prophylactic treatment which prevents an infection by H. pylori (for example immunization) can be used as well as a treatment which eliminates an infection by H. pylori that is already present to treat these frequent gastroduodenal diseases. In addition to certain higher primates, it is previously known that humans are the only natural host of H. pylori. The relatively recent discovery that the domestic cat can also be infected with H. pylori sheds new light on the manner of transmission and possible reservoir of the bacterium outside the human organism. Occasionally successful H. pylori culture of feces of infected people and the ability of bacteria to survive for months in water supports the hypothesis of fecal-oral transmission. It is also considered as probable oral-oral direct transmission based on family studies. The infection usually occurs in childhood within the family, with precarious living conditions and a poor hygiene standard which correlate positively with the frequency of infection. After oral uptake the bacteria first reaches the extremely acid lumen of the stomach (pH 1-2). Here it becomes possible the survival of the bacteria by the production of the urease enzyme which leads to the breakdown of urea that is present and therefore to local neutralization of the acid pH value in the stomach. By means of chemotactic orientation and flagella-dependent motility, the microorganisms move to mucosal layers buffered with bicarbonate from the region of the antrum of the stomach, which is, in fact, their natural habitat. Here, they are in a unique ecological niche which, due to the acid barrier, is accessible only by some bacterial species that compete with the microorganism. Probably the microorganisms are oriented to itself by means of a gradient of pH between the lumen (pH 1-2) and the surface of epithelial cells (pH 6-7) in order to reach the epithelium. Due to its spiral shape, its motility in viscous mucous membranes, the production of mucosal modifying enzymes and finally its microaerophilic way of life, these germs adapt optimally to the living conditions of this habitat. They usually spend their time in deep crypts in the antrum region where they are protected from external influences such as, for example, acid, pepsin and also some forms of medicine for their eradication such as, for example, antibiotics. Part of the population (ca. 20%) is closely associated with epithelial cells, especially with mucosal producing cells. Under the condition of a gastric metaplasia, that is, the acid-induced formation of gastric epithelium in the duodenum, the metaplastic areas in the duodenum are also colonized which generates the prerequisites for the development of a duodenal ulcer (ulcus duodeni). A complete expression of Helicobacter with the mucosa, dissemination is probably avoided by its ability to adhere, so that the bacteria can persist for years, decades or even throughout life (chronic infection).

Before the consistency and importance of H. pylori for ulcerative diseases was known, they were treated with what were called antacids or H2-receptor antagonists. These were substances which inhibit the acid secretion of the parietal cells of the stomach. The action of these pharmaceutical agents usually leads to the healing of ulcers but, since one of the causes of ulcers, ie H. pylori infection is not eliminated by it, in most cases recurrence occurs of ulceration (relapse) in a short period of time. A therapy often used in ulcerations is a treatment with bismuth. Several bismuth salts (CBS, BSS) have a bactericidal effect on H. pylori. However, a total eradication of these germs is only obtained in 8-32% of cases. Apparently, the treatment leads to a temporary suppression of the germs but after stopping the treatment the infection re-fluoresces again in most of the cases. Long-term therapy with high doses leads to accumulation of the substance in the liver, kidney and nervous system and has considerable neurological side effects (Malfertheiner, 1994).

Since it has been discovered that gastroduodenal ulcer diseases are infectious diseases, one goal of treatment is to eradicate the pathogens with antibiotics. The monotherapy with various antibiotics (amoxicillin, nitrofuran, furazolidine, erythromycin a .. o.) However, has not been satisfactory since even in this case, the eradication of germs only occurs between 0-15% of cases. So far, the most successful treatment is obtained by a combination of an acid blocker (Ompeprazole) with an antibiotic (Amoxicillin) which leads to eradication rates above 80% (Malfertheiner, 1994). However, antibiotic treatment to eliminate H. pylori is not promising as a long-term solution since the assumption that the bacteria will rapidly develop resistance to antibiotics must be established. Therefore, there is a need for new forms of therapy to control H. pylori infection and in particular for vaccines which are specifically directed against H. pylori virulence factors. The virulence factors denote the properties of a pathogenic bacterium which allows to colonize a particular ecological niche in the host's body and multiply in that place despite the immune response and the non-specific defense mechanisms of the host organism. Knowledge about the factors of violence therefore helps in a better understanding of the course and mechanisms of an infectious disease. The most important virulence factors previously examined for H. pylori are urease, flagella and adesines in the production of a cytotoxin. The urease and the enzyme on the surface of the bacterium are made up of two subunits (UreA, 26 kDa, UreB, 66 kDa) which constitute up to 5% of the total bacterial protein. Urease separates or breaks down urea which is found at low concentrations in the gastric juice and breaks it down into ammonia and carbon dioxide. According to the current perception, the bacteria surround themselves with a cloud of ammonia which leads to a local neutralization of the acid in the gastric juice. The extremely high utility of the bacterium can be attributed to the presence of polar flagella which allow the bacteria to move in the viscous mucus of the gastric mucous membrane and therefore reach the layers of epithelial cells. The group of genes for ureases (ureA-ureH) as well as the genes for the formation of flagella (flaA, flaB) in E. coli have been cloned and isogenic mutants have been sequenced and constructed. Approximately 50-60% of all isolated H. pylori strains produce an 87 kDa protein called vacuolizing cytotoxin, which induces the formation of cytoplasmic vacuoles in cell cultures in vitro. The vacA gene which codes for the cytoxine of H. pylori has also been cloned and at the same time has been genetically characterized. In addition, it is assumed that the cytotoxin-producing strains have a higher pathogenic potential compared to strains that do not produce this toxin. In addition, a positive correlation has been found between the production of the cytotoxin and the development of gastric ulcers. Research regarding the adherence of H. pylori to epithelial cell lines in vitro shows that the bacterium can bind to many cell lines of different tissues. In contrast, H. pylori shows a very pronounced selective adherence to species and tissue (tropism) in the host organism. Therefore, the bacterium is only attached to epithelial cells which are of the gastric type of epithelial cells. This selectivity is explained by a specific interaction between a bacterial adesine and a specific cellular receptor. Until now, several potential adhesins of H. pylori have been described and a gene (hpaA) has been cloned and sequenced, which codes for what is called hemagglutinin that binds N-acetylneuraminilactose (Evans et al., 1993). This is a protein which would be able to recognize a receptor that contains cyhalic acid on epithelial cells. However, the significance of this adhesin for H. pylori infection is controversial. Other potential adhesins are characterized only by their molecular weight or by their receptor binding specificity. These include a 63 kDa protein which appears to be homologous to the S exoenzyme of Pseudomonas aeruginosa, an adhesin with ADP-ribosyl transferase activity. In addition, it is suspected that there is an unidentified adhesin hitherto which mediates a specific binding to the Lewisb blood group antigen of gastric epithelial cells (Falk et al., 1993).; Borén et al., 1993). Infection with H. pylori leads to a chronic inflammatory reaction of the gastric mucosa (gastritis). In addition, it is induced to a specific systemic immune response against H. pylori antigen; however, the formation of secretory antibodies in the stomach (slgA) has not yet been fully elucidated. As a result of inflammation, various immune cells are present in the gastric mucosa and submucosa, for example, polymorphonuclear leukocytes, monocytes, macrophages, lymphocytes and plasma cells (Blaser, 1992). In addition, H. pylori activates neutrophils as well as monocytes and macrophages in vitro (Mai et al., 1991). Experiments with specific antibodies and complements show a rapid inactivation of H. pylori by neutrophils in vitro. However, in the in vivo situation these mechanisms do not lead to an inactivation of the pathogenic bacteria. The way in which H. pylori survives for a prolonged period in the host is not clear, although it activates the defense mechanism mentioned in the above. The host is not able to resist H. pylori infection under natural conditions. Therefore, it has been more surprising that urease, an essential virulence factor of H. pylori (see above), has a great potential as a vaccine (US patent application USSN 07 / 970,996 Urease-based vaccine against Helicobacter Infection) . In the Helicobacter felis / mouse model (H. felis is a species of Helicobacter which naturally colonizes the cat's stomach and also infects the mouse (It is possible to demonstrate that oral vaccination of H. pylori urease or recombinant urease B subunit (rUreB) can protect mice against infection by H. felis (preventive vaccine) and can also eliminate an infection which is already present (therapeutic vaccine) (Michetti et al., 1994; Corthesy-Theulaz et al., Gastroenterol., In print). A decisive factor in oral vaccination is the use of adjuvants such as, for example, cholera toxin which, among others, seems to be important in converting the immune reaction from the production of systemic antibodies to secretory antibodies. The objective of the present invention is to provide a method by which the secretory genes of Helicobacter pylori, which are potential candidates for vaccines can be identified easily and quickly. This objective is obtained by a method to identify Helicobacter pylori secreting genes in which (a) a gene bank of H. pylori DNA is elaborated in a host organism which contains an inducible transposon coupled to a marker for secretory activity, (b) insertion of the transposon into the H. pylori DNA is induced, and (c) the marker is used to select clones which contain a secretory gene. The term "secretory gene" or "gene with secretory activity" is intended to indicate a gene which codes for a secretory polypeptide, i.e., for a polypeptide exported from the cytoplasm.

The method according to the invention is particularly suitable for identifying H. pylori adhesion genes in which, in addition, (d) a retransformation of H. pylori is carried out with the DNA of the clones of the gene bank, preferably with clones which contain genes with secretory activity, which produces isogenic H. pylori mutant strains by integration of the DNA into the chromosome, and (e) adherent deficient H. pylori mutant strains are selected. The method according to the invention allows the identification of adhesion genes and the adhesion proteins (adhesins) encoded by them from H. pylori and those which are responsible for the specific interaction of the bacterium with epithelial cells of the gastric mucosa. Since H. pylori adhesins are usually produced only in very low amounts, it is usually extremely difficult to identify and isolate them by biochemical means. In addition, due to the tendency of H. pylori to spontaneously autolish, the cytoplasmic proteins bind to the bacterial surface. In a biochemical purification, such proteins have been mistakenly identified as adhesins in the past (Doig et al., 1992; Doig et al., 1993). The method according to the invention allows quick and easy identification of the adhesins. In this method, individual genes are inactivated in the H. pylori chromosome with the help of a transposon and mutants are produced which have effects on different chromosomal gels. Mutants of these independent defective mutants can be selected by specific selection of those mutants which are activated by adhesin genes and therefore are not capable of binding to receptors of the corresponding target cells. The first step in the method according to the invention comprises establishing a gene bank of H. pylori DNA in a host organism, preferably gene bank plasmid in a prokaryotic host organism such as E. coli. Subsequently, cloned H. pylori genes are mutagenized with the help of a transposon which is coupled to a marker for secretory activity. A particularly preferred transposon is the TnMax9 transposon which was deposited on 05.26.95 in the "Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSM)", Mascheroder Weg lb, De-38124 Braunschweig in E. coli E 181 under the name of the file DSM 10008, in accordance with the Budapest contract rule. Figure 1 shows a schematic representation of TnMax9. TnMax9 transports a copy of a gene for S-lactamase without the promoter and a signal sequence (blaM) as a marker directly after the repeated and inverted (IR) sequences that delimit the current transposon (Tadayyon and Broome-Smith, 1982). After insertion of TnMax9, the blaM gene is fused to the target gene and when the inactivated target gene is expressed, a fusion protein is produced between the target X gene and blaM if the transposon is inserted in the correct orientation and in the correct reading frame with respect to the X gene. Provided that the inactivated X gene codes for the secretory protein, in particular for a protein exported by the sec-dependent transport path (Pugsley, 1993), the fusion protein is It will transport through the cytoplasmic membrane of the host cell E. coli into the periplasm. Here, the fusion protein exhibits its activity (rupture of 0-lactam antibiotics) and therefore mediates a resistance of the E. coli clone corresponding to the antibiotic ampicillin. However, if the X gene is a gene for a cytoplasmic protein it can also result in a fusion protein that depends on the insert (orientation, frame) but does not show any β-lactamase activity since 0-lactamase is only activated in the periplasmic environment but not in the cytoplasm. Therefore, this method can be used to identify specific mutants in genes which code for exported proteins by examining the mutants to determine 0-lactamase activity after transposon mutagenesis in E. coli. Since the genes for H. pylori adhesin that are sought must also be export proteins, a special collection of H. pylori transposon mutants in genes which code for exported proteins can greatly reduce the number of mutants that must be exported. try on In this way, the method according to the invention provides a highly enriched mutant bank which can avoid a search of a conventional mutant bank with ca. 2000 - 4000 clones which is very difficult to carry out in a practical way in a laboratory. Most of the chromosomal genes of a bacterium that codes for cytoplasmic proteins can be eliminated from the beginning by the selection step directed towards the identification of secretory genes. Therefore, the number of H. pylori mutants that must be tested on particular target cells for the loss of their ability to adhere can be reduced to an acceptable level for practical purposes. The examination of bacterial adhesion mutants can be carried out at various levels. Usually, epithelial cell lines are used for this, to which bacteria specifically bind. In the case of cell lines of special gastric carcinoma is H. pylori can be used, for example, the cell line KatoIII (ATCC HTB 103) which has already been described by various authors as a model of adherence (Clyne and Drumm , 1993). Although cell lines are relatively simple to handle and can usually be cultured in large numbers in vitro, it is not possible to fundamentally exclude changes in the quantitative and qualitative expression of surface proteins such as, for example, receptors, due to immortalization of the cells. Therefore, tissue models are an alternative for this. In this case, ultrathin sections are prepared from fixed tissue (microtome) which is incubated after saturation with the bacteria. Bacteria which do not bind to the receptors are removed by washing and the bound bacteria can be visualized with dyes (fluorescent dyes or special bacterial dyes). As a third stage, animal models can be used. These are in vivo experiments which can only be carried out if animal models suitable for the microorganism are available. The test for determining defective mutants in H. pylori adhesion is preferably carried out with the help of the KatoIII cell line. For this, the mutants can be grown on agar plates and subsequently labeled with the fluorescent dye fluorescein isothiocyanate (FITC). Subsequently, the labeled bacteria are added to epithelial cells incubated for 1 h at 37 ° C. A fluorescent microscope is then used to examine whether individual mutants are still able to bind to the epithelial cell line. In a preferred embodiment of the method according to the invention, a gene bank of H. pylori is made in a minimal plasmid vector which allows a selection for a transposon insertion in the vector DNA. The use of such minimal plasmid vectors greatly improves the efficiency of the mutagenesis process due to the size of the generic elements of the plasmid. In addition, an insertion of the transposon into the vector DNA which essentially only consists of the elements necessary for replication, selection and cloning, probably leads to loss of the ability of the vector to propagate in the host cell. Furthermore, it is preferable to constitute a gene bank of H. pylori in a plasmid vector which contains a sequence suitable for transfer and conjugated in other host organisms. An example of such oriT sequence (Fürste et al., 1989) the presence of which avoids the tedious isolation of plasmid DNA after inducing the transposon and the formation of suitable receptor cells. The presence of an expression signal in the plasmid vector to which it is inserted into H. pylori DNA that is operatively linked is also preferred. In this way, it is possible to transcribe sequences of H. pylori genes which could not otherwise be transcribed due to the presence of specialized promoter sequences. In addition, it is also possible to transcribe genes which are organized in operons which can not be cloned as a complete unit. The use of a weak promoter is preferred since stronger promoters can lead to slow transformation rates and even to deletions and rearrangements of the cloned H. pylori sequences. A particularly preferred plasmid vector is pMin2 (DSM 10007) which was deposited on 05.26.95 in E. coli strain DH5or according to the rules according to the Budapest contract. Figure 2 shows schematically the steps used to construct pMin2. As already mentioned, it is preferable for the method according to the invention that a conjugative transfer of the plasmid DNA from the host organism to a recipient organism be carried out which allows the selection of plasmids containing transposons to select clones which contain a secretory gene. The selection for adhesion genes is preferably carried out by retransformation of H. pylori by means of which a mutant strain is produced by integration of DNA into the chromosome. Before this retransformation the marker gene for example, the BlaM gene is preferably deleted in order to obtain a higher transformation rate, to avoid the production of H. pylori strains capable of producing / S-lactamase and to avoid possible interferences of the bacterial export apparatus by BlaM fusion proteins. A further subject matter of the present invention is a gene bank of H. pylori DNA in a host organism which contains fragments of H. pylori DNA inserted into a vector characterized in that the genome of H. pylori is fully presented with a probability greater than 90% and that the host organism also contains an inducible transposon coupled to a marker for secretory activity. With a genome size of H. pylori of 1.7 Mb (Bukanov and Berg, 1994) and an average insert size of, preferably 3-6 kb especially preferably for ca. 4 kb, the gene bank according to the invention preferably comprises at least 400 clones per genome. Particularly preferably, the gene bank comprises 2000 to 4000 clones. H. pylori DNA is preferably inserted into a plasmid vector, preferably especially in a minimal plasmid vector as defined above. The host organism in which the gene bank is preferably located is a bacterium, particularly preferably a gram-negative bacterium and more preferably E. coli. The gene bank according to the invention can be used to identify secretory genes and, in particular, adhesion genes of Helicobacter pylori. It is possible to identify a gene for adhesin from H. pylori called alpA and a polypeptide encoded by this gene using the method according to the invention. Therefore, a subject matter of the present invention is also a DNA molecule which comprises: (a) a nucleotide sequence as shown in SEQ. FROM IDENT. N0: 1 (b) a nucleotide sequence which corresponds to the sequence according to (a) within the scope of the degeneracy of the genetic code, or (c) a sequence of nucleotides which hybridizes to the sequences according to ( a) or / and (b) under conditions of restriction. In addition to the nucleotide sequence shown in SEC. FROM IDENT. NO: l and a nucleotide sequence corresponding to this sequence within the scope of the degeneracy of the genetic code, the present invention also encompasses a DNA sequence which hybridizes to one of these sequences under restriction conditions. In accordance with the present invention, the term "hybridization" is used as described by Sambrook et al. (Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory Press (1989), 1101-1104). According to the present invention, a hybridization under restriction conditions means that a positive hybridization signal is still observed after washing for 1 h with 1 x SSC and 0.1% SDS at 55 ° C, preferably at 62 ° C and especially preferably at 68 ° C, in particular for 1 h in 0.2 x SSC and 0.1% SDS at 55 ° C, preferably at 62 ° C and especially preferably at 68 ° C. The present invention encompasses a nucleotide sequence under such washing conditions that hybridizes to one of the nucleotide sequences shown in SEQ. FROM IDENT. NO: l or with a corresponding nucleotide sequence within the scope of the degeneracy of the genetic code. The DNA molecule according to the invention preferably encodes a polypeptide having the ability to adhere to human cells, in particular human gastric epithelial cells. Furthermore, it is preferred that the DNA molecule according to the invention have a nucleotide-level homology of at least 70%, particularly preferably at least 80%, with the nucleotide sequence shown in the SEC. . FROM IDENT. NO: l. It is further preferred that the DNA molecule has a length of at least 15 and preferably at least 20 nucleotides. A further subject matter of the present invention is a vector which contains at least one copy of a DNA molecule according to the invention. This vector can be any prokaryotic or eukaryotic vector on which the DNA sequence according to the invention is located, preferably under the control of an expression signal (promoter, operator, extender, etc.). Examples of prokaryotic vectors are chromosomal vectors such as, for example, bacteriophages (for example bacteriophage?) And extrachromosomal vectors such as plasmids, whereby circular plasmid vectors are especially preferred. Suitable prokaryotic vectors are described, for example, by Sambrook et al., Supra, Chapters 1 to 4. On the other hand, the vector according to the invention can also be a eukaryotic vector, for example, a yeast vector or a vector suitable for higher cells (for example a plasmid vector, a viral vector, a plant vector). Such vectors are described, for example, by Sambrook et al.; Supra, chapter 16. A further subject matter of the present invention is a cell which is transformed with a vector according to the invention. In a preferred embodiment, the cell is a prokaryotic cell, preferably a negative prokaryotic cell, particularly preferably an E. coli cell. However, on the other hand, the cell according to the invention can also be a eukaryotic cell such as a fungal cell. (for example, yeast) an animal or a vegetable cell.

The invention also relates to a polypeptide which is encoded by a DNA molecule according to the invention. Preferably, the polypeptide is capable of adhering to human cells and comprises: (a) the amino acid sequence shown in SEQ. FROM IDENT. NO: 2, or (b) an amino acid sequence that immunologically cross-reacts with the sequence according to (a). Preferably, the polypeptide according to the invention has a homology of at least 80%, and more preferably at least 90% with respect to the amino acid sequence shown in SEQ. FROM IDENT. NO: 2. The polypeptides according to the invention, preferably produced by transforming a cell with a DNA molecule or a vector according to the invention, culturing the transformed cell under conditions in which expression of the polypeptide takes place. and isolating the polypeptide from the cell or / and the culture supernatant. In this process, the polypeptide according to the invention can be obtained as a fusion polypeptide as well as a non-fusion polypeptide. The polypeptide according to the invention can also be used as an immunogen to produce antibodies. Therefore, the present invention also relates to an antibody which is directed against a polypeptide according to the invention. Preferably, the antibody is directed against the N-terminal part, eg, the first 250 'amino acids of the amino acid sequence shown in SEQ. FROM IDENT. N0: 2 A further aspect of the present invention relates to a pharmaceutical composition which contains a DNA molecule according to the invention, a polypeptide according to the invention or an antibody according to the invention as an active substance optionally together with auxiliary substances. , diluents, additives and common pharmaceutical carriers. The pharmaceutical composition according to the invention can be used, on the one hand, to diagnose an infection by Helicobacter pylori. The diagnosis at the nucleic acid level is preferably carried out using hybridization probes which contain a DNA according to the invention which is specific for the alpA gene or by amplification using DNA molecules according to the invention as primers. At the protein level, the diagnosis is preferably carried out with the aid of antibodies according to the invention. On the other hand, the pharmaceutical composition can also be used to prevent or combat a Helicobacter pylori infection. For therapeutic application, the polypeptide or parts thereof are used to produce an active vaccine or the antibody is used to produce a passive vaccine. It is considered to further elucidate the invention by the following examples and figures. Figure 1 shows a linear restriction map of the TnMax9 transposon (DSM 10008) which is used to identify and inactivate the alpA gene. Figure 2 shows a schematic representation of the construction of the minimal plasmid vectors pMinl and pMin 2 (DSM 10007) which are suitable for efficient transposon mutagenesis. Figure 3A shows the principle of transposon mutagenesis selection with Tnmax9 and the scheme for producing H. pylori mutants which have a defect in the secretory gene. Figure 3B shows a schematic representation of the method for identifying mutants of H. pylori which have a defect in a secretory gene. Figure 4 shows a restriction map of plasmid pMul40 which contains the regulatory region and the 5 'end of the alpA gene (SEQ ID NO: 1). The alpA gene is inactivated by the TnMax9 transposon insertion (see triangle marked TnMax9). When the plasmid is expressed, an alpA-β-lactamase fusion protein is obtained. The original clone is pMul40 from the mutant gene bank, from which a defective H. pylori strain can be obtained in adhesion of 1-140 by transformation and homologous recombination. Figure 5 shows an immunoblot of total cell lysates of H. pylori of the wild-type strain 69A (1) as well as isogenic mutant strains Pl-140 (2) and Pl-179a (3). The wild-type strain 69A contains the alpA protein with a molecular weight of ca. 53 kDa. The mutants alp Pl-140 and Pl-179a do not have the AlpA protein. Immunoblotting is carried out using AB 202 antibody which is directed against the N-terminal part of the recombinant AlpA protein. The SEC. FROM IDENT. NO: l shows the nucleotide sequence of the adhesion gene of H. pylori alpA and the corresponding amino acid sequence. The SEC. FROM IDENT. NO: 2 shows the amino acid sequence of the AlpA adhesion polypeptide of H. pylori.

Example 1 Construction of minimal vectors Plasmid pRH144 is constructed by ligation of the ColEl origin of replication (orioni.-.) And the tetracycline resistance gene (Tet), which are applied as PCR fragments from pBR322 using the primer pairs RH104 ( 5 '-AGC TGA ATT CAT GTT TGA CAT TGC CAT ATA GAT GAG CTT TAA TGC GGT AGT T-3') and RH105 (5 '-AGC TCT GCC GCC GCC GGC TTC CAT TCA G-3') and RH106 (5 ' -AGC TCT GCA GAG ATC AAAGGA TCT TCT T-3 ') and RH107 (5' -TCT AGA ATT CGT ATC AGC TCA CTC AAA G-3 '). PRH146 is constructed by inserting an oriT fragment which is obtained from plasmid RP4 (Marshall et al., 1984) by PCR amplification using the initiator pair DF001 (5 '-GTA CTG CAG CTT GGT TTC ATC AGC CA-3') and DF 002 (5 '-GTA CTG CAG TTC AGT AAT TTC CTG CAT-3'). Subsequently, the multiple cloning site of plasmid pIC20R1 (Thomas, 1981) is inserted into the EcoRI site of pRH146. In order to obtain the pMinl plasmid, the polylinker pRHl46 is replaced by a double-stranded synthetic polylinker region which is cloned into the EcoRI site of pRH146 and is composed of the two partially complementary oligonucleotides RH117 (5 '-AAT TAG ATC ATT AAA GGC TCC TTT TGG AGC CTT TTT TTT TGA ATT CAG ATC TCG AGG TAC CCG GGA TCC TCT AGA-3 ') and RH118 (5' -AAT TAG ATC AAA AAA AAA GCC CGC TCA TTA GGC GGG CTA AGC TTG TCG ACÁ TCG ATC TAG AGG ATC CCG GGT ACC-3 ').

The flanking EcoRI sites are destroyed by cloning procedure. The cloned polylinker region contains the transcription terminators of phage fd (terfd) and the trpA gene (tert? PA). In order to obtain the pMin2 plasmid, the gene promoter (Piga) derived from gonococci is amplified as a PCR fragment from the plasmid pIP 100 (Pohlner et al., 1987) using oligonucleotides SO009 (5 '-GGA TCC GAA TTC TCA TGT TTG ACA G-3') and SO010 (5 '-GTC GAC AGA TCT TTT AAT AGC GAT AAT GT-3'). The amplification fragment is ligated into the EcoRI and BglII sites of pRHl60. The trpA terminator is separated by substitution of the BglII / BamHI fragment of pMinl by the corresponding fragment of pRH146. Figure 2 shows the construction scheme of pMinl and pMin2. E. coli, strain DH5a containing the pMin plasmid in DSM was deposited under the file number 10007.

Example 2 Construction of the TnMax9 transposon The plasmid pRHHO which is a derivative of pTnMaxl (Haas et al., 1993) with a linearized pIC20R2 vector with SalI at the cellular Sali cleavage site is used as a basis for the construction of pTnMax9.

Subsequently the central region of TnMaxl is deleted (ori ^ -res-catoc) by rupture and religation with HindIII. The sequences for the forward sequencing initiator M13 (M13 initiator) and inverse M13 (M13-RP1) are introduced (ligation) at the HindIII cleavage site of pRHllO by insertion of two partially complementary oligonucleotides RH096 (5 '-AGC TTA CTG GCC GTC GTT TTA CAG CGG CCG CAG GAA AT GCT ATG ACC GA-3') and RH097 (5 '- AGC TTC GGT CAT AGC TGT TTC CTG CGG CCG CTG TAA AAC GAC GGC CAG TA-3 '). The resulting plasmid pRH140 contains a unique Notl restriction cleavage site between the two initiator binding sites. The Tnl72l DNA fragment is amplified from the plasmid pJOE106 (Schofff et al., 1981) with the help of the oligonucleotide initiator pair (RH098 (5'-AGA AGC GGC CGC AAA AGG ATC CAT AGG TGC AAG CAA GTT A -3 ') and RH099 (5' -AGC TGC GGC CGC AAA AAG ATC TCA AAG CCC ATT TCT GTC AGG-3 ') and inserted into the Notl cleavage site pRH140 As a result, the plasmid pRH141 is formed which has unique restriction cleavage sites BamHl and BglII to the left and right of res.The origin of replication (orifd) is now isolated together with the resistance section cat-l from pRH42 as a BamHl fragment and ligated into the BamHl cleavage site of pRH141 The deletion of the pIC20R2 vector (with Sali) finally leads to pTnMaxd as a base construct for all subsequent TnMax transposonee., the gene for S-lactamase without a promoter and without a signal sequence (blaM gene) is amplified from plasmid pBR322 (Sutcliffe, 1979) by means of PCR with the help of oligonucleotides RH124 (5 '-AGT TGC GGC CGC ACC CAG AAA CGC TGG TG-3 ') and RH127 (5'-AGC TAG ATC TAG ATT ATC AAA AAG GAT C-3') and inserted into the Notl and BglII cleavage sites of TnMax5, which results in pTnMax7 . The Notl rupture site between catheter and the inverted repeated sequence (IR) of pTnMax7 is separated by filling the superimposed ends with Klenow polymerase. The BglII cleavage site at the 3 'end of blaM is converted to a Notl cleavage site by insertion of the complementary oligonucleotide pair SO012 (5' -GAT CA GTC GCG GCC GCC TGA T-3 ') and SO013 (5' - GAT CAT CAG GCG GCC GCG ACT T-3 ') which leads to the derived transposon pTnMax9. E. coli, strain E181, which contains the transposon derived pTnMax9 is deposited in DSM under the file number 10008. 3 Construction of a gene bank of the H. pylori plasmid. A plasmid gene bank was prepared from the chromosomal DNA of wild-type H. pylori strain 69A. For this, chromosomal DNA is isolated by the method of Leying et al. (1992) from H. pylori and partially separated with each of the restriction endonucleases Sau3AI and Hpall. Subsequently, the DNA fragments are separated on preparative agarose gel and 3-6 kb fragments are eluted from the gel. These DNA fragments are ligated into the plasmid vector pMin2 which is constructed especially for this purpose and which has been separated with the restriction enzymes BglII and Clal, ligated (T4 ligase) and the ligation mixture transformed into E coli strain E 181, which is a derivative of strain HB101 (Bayer and Roulland-Dussoix, 1969) that contains the phage lysosomes? XCH616 and has already been transformed with the TnMax9 transposon. In this process, ca. 2400 independent transformants.

Example 4 Isolation of H. pylori mutants In FIGS. 3A and 3B, the principle of selection of mutagenesis and the method for identifying mutants are shown schematically.

In order to carry out the mutagenesis of the transposon, 10 transformants are accumulated in each case and mutually induced by means of a total of 190 accumulated, each with 10-20 clones that are treated additionally. 191 mutant clones of ampicillin-resistant E. coli plasmids which independently transport mutated H. pylori genes are isolated from this mutagenesis. These 192 plasmids are isolated from E. coli and used for the retransformation of H. pylori strain 69A. 135 mutants of H. pylori are isolated from these 192 transformations which are probably mutated in genes which code for secretory proteins. The collection of mutant H. pylori is then tested in a screening assay for H. pylori mutants which have lost their ability to bind to KatoIII epithelial cells. For this, the mutants are labeled with FITC and cultured for 1 hour at 37 ° C along with the epithelial cells. The test for adherence is carried out directly by observation with a fluorescence microscope. In this case there are two mutants (numbers Pl-140 and Pl-179a) which show a greatly reduced adherence. Both mutants also do not show adherence in the second adhesion model, sections of human stomach tissue. H. pylori wild-type strain as well as all other mutants also show strong adherence in this model. The plasmid pMul40 used to produce the mutant strain Pl-140 is shown in Figure 4. Plasmid pMU179a (not shown) is used to produce the mutant strain Pl-179a. The independent transformations of both plasmids in H. pylori 69A lead to an identified adhesion defect which shows that secondary mutations have not occurred in the bacterial chromosome but rather in the insertion of TnMax9 in the cloned adhesin gene which leads to the phenotype observed of H. pylori mutants. The mapping and sequencing of the genes of the clones of plasmid pMul40 and pM179 inactivated by the transposon TnMax9 shows that both clones are the same gene, - the transposon only inserts in different sites. Since the encoded protein is a lipoprotein, that is, a protein which has fixed to the membrane with a lipid anchor, the corresponding gene is called alpA (adhesion-associated lipoprotein A). Data from predictions of secondary structure by computer of membrane proteins and certain protein sequence conserved in the C-terminal part of the protein (C-terminal phenylalanine, Struyvé et al., 1991); argullen in favor of an integral membrane protein incorporated from the outer membrane of gram-negative bacteria. 5 Expression of the alpA gene and production of antibodies Subsequently, a fusion protein is produced with the help of the expression system pEV40 E. coli (Pohlner et al., 1993) which comprises ca. 50% of the N-terminal part of AlpA and the N-terminal histidine tag. This is used to purify the fusion protein by chelate affinity chromatography with a Ni2 agarose column and is used to obtain a mouse antiserum. This antiserum, which is designated AK202, recognizes the AlpA protein as a 53 kDa protein in an immunoblot of a total cell lysate of H. pylori. As expected, the 53 kDa AlpA protein is not detectable in an immunoblot in the case of H. pylori, mutants Pl-140 and Pl-179a (see Figure 5). The proof that AlpA is a lipoprotein is carried out by a specific palmitylation of the fusion protein AlpA-jS-lactamase which is formed by the original insertion of TnMax9 in alpA.

LITERATURE Alper, J. (1993) Ulcers as an infectious disease. Science 260: 159-160. Bizzozero, G. (1893) Ueber die schlauchfórmigen Drusen des Magendarmkanals und die Beziehungen ihres Epithels zu dem Oberfláchenepithel der Schleimhaut. Arch Mikr Anast 42: 82. Blaser, M.J. (1992) Hypothese on the pathogenesis and natural history of Helicobacter pylori induced inflammation. Gastroenterol 102: 720-727. Borén, T., Falk, P., Roth, K.A., Larson, G., Normark, S. (1993) Attachment of Helicobacter pylori to gastric epithelium mediated by blood group antigens. Science 262: 1892-1895. Boyer, H.W. and Roulland-Dussoix, D. (1969) A complementation analysis of the restriction and modification of DNA in Escherichia coli. J. Mol. Biol. 41: 459-472. Bukanov, N.O., Berg, D.E. (1994) Ordered cosmid library and high-resolution physical-genetic map of Helicobacter pylori strain NCTC11638. Mol Microbiol 11: 509-523. Clyne, M., Drumm, B. (1993) Adherence of Helicobacter pylori to primary human gastrointestinal cells. Infect Immun 61: 4051-4057.

Doig, P., Austin, J.W., Kostrzynska, M., Trust, T.J. (1992) Production of a corserved adhesin by the human gastroduodenal pathogen Helicobacter pylori. J. Bacteriol 174: 2539-2547. Doig, P., Austin, J.W. , Trust, T.J. (1993) The Helicobacter pylori 19.6-kilodalton protein is an intron-containing protein resembling ferritin. J. Bacteriol 175: 557-560. Evans, D.G., Karjalainen, T.K., Evans, D.J., Jr., Graham, D.Y., Lee, C.-H. (1993) Cloning, nucleotide sequence, and expression of a gene encoding an adhesin subunit protein of HelicoJacter pylori. J. Bacterial 174: 674-683. Falk, P., Roth, K.A., Borén, T., Westblom, T.U., Gordon, J.I., Normark, S. (1993) An in vitro adherence assay reveals that Helicobacter pylori exhibits cell lineage-specific tropism in the human gastric epithelium.

Proc. Nati Acad. Sci USA 90: 2035-2039. Fürste, J.P., Pansegrau, W., Ziegelin, G., Krdger, M. and Lanka, E. (1989) Conjugative transfer of promiscuous IncP plasmids: interaction of plasmid-encoded products with the transfer origin. Proc. Nati Acad. Sci USA 86: 1771-1775. Goodwin, C.S., Armstrong, J.A., Chilvers, T., Peters, M., Collins, M.D., Sly, L., McConnell, W., Harper, W.E.S. (1989) Transfer of Campylobacter pylori and Campylobacter mustelae to Helicobacter gen. nov. as Helicobacter pylori comb. nov. and Helicobacter mustelae comb. nov., respectively. Int. J. Syst. Bact 39: 397-405. Haas, R., Kahrs, A.F., Facius, D., Allmeier, H., Schmitt, R., Meyer, T.F. (1993) TnMax- a versatile mini-transposon for the analysis of cloned genes and shuttle mutageneeis. Gene 130: 23-31. Isaacson, P.G., Spencer, J. (1993) Is gastric lymphoma an infectious disease? Hum. Pathol. 24: 569-570. Lee, A., Fox, J., Hazell, S. (1993) Pathogenicity of Helicobacter pylori: a perspective. Infect. Immun. 61: 1601-1610. Leying, H., Suerbaum, S., Geis, G., Haas, R. (1992) Characterization of flaA, to Helicobacter pylori flagellin gene. Mol. Microbiol 6: 2863-2874. Mai, U.E.H., Perez-Perez, I., Wahl, L.M., Wahl, S.M., Blaser, M.J., Smith, P.D. (1991) Soluble surface proteins from Helicobacter pylori actívate monocytes / macrophagee by lipopolysaccharide-independent mechanism. J. Clin Invest. 87: 894-900. Malfertheiner P. (1994) Helicobacter pylori - Von der Grundlage zur Therapie. Bayerdorfer E., Birkholz S., Bdrsch G. et al., (Eds.) Stuttgart, New York: Georg Thieme Verlag; p. 1-104.

Marsh, J.E., Erfle, M. and Wykes, E.J. (1984) The pIC plasmid and phage vectors with versatile cloning sites for recombinant selection by insertional inactivation. Gene 32: 481-485. Marshall, B.J., Royce, J., Annear, D.I., Goodwin, C.S., Pearman, J.W., Warren, J.R., Armstrong, J.A. (1984) Original isolation of Campylobacter pyloridis from human gastric mucosa. Microbes Lett. 25: 83-88. Michetti, P., Corthesy-Theulaz, I., Davin, C, Haas, R., Vaney, A.C., Heitz, M., Bille, J., Kraehenbuhl, J.P., Saraga, E. # Blum, A.L. (1994) Immunization of Balb / c mice against Helicobacter felis infection with Helicobacter pylori urease. Gastroenterol 107: 1002-1011. Nomura, A., Stemmermann, G.N., Chyou, P.H., Kato, I., Perez-Perez, G.I., Blaser, M.J. (1991) Helicobacter pylori infection and gastric carcinoma among japanese Americans in Hawaii. N Engl. J. Med. 325: 1132-1136. Parkin, D.M., Láárá, E., Muir, C.S. (1988) Estimates of the worldwide frequency of six major cancers in 1980. Int. J. Cancer 41: 184-197. Parsonnet, J., Friedmann, G.D., Vandersteen, D.P., Chang, Y., Vogelman, J.H., Orentreich, N., Sibley, R.K. (1991) Helicobacter pylori infection and the risk of gastric carcinoma. N. Engl. J. Med. 325: 1227-1231.

Pohlner, J., Kramer, J. and Meyer, T.F. (1993) A plasmid system for high-level expression and in vitro processing of recombinant proteins. Gene 130: 121-126. Pohlner, J., Halter, R., Beyreuther, K. and Meyer, T.F. (1987) Gene structure and extracellular secretion of Neiseeria gonorrhoeae IgA protease. Nature 325: 458-462. Pugsley, A.P. (1993) The complete general secretory pathway in gram-negative bacteria. Microbiol Rev. 57: 50-108. Schdffl, F., Puhler, A., Altenbuchner, J. and Schmitt, R. (1981) The tetracycline resistance transposons Tnl721 and Tnl77l have three 38-base-pair repeats and genrerate five-base-pair repeats. Mol. Gen. Genet 181: 87-94. Sipponen, P. (1992) Natural history of gastritis and its relationship to peptic ulcer disease. Digestion 51: 70-75. Sipponen, P., Hyvárinen, H. (1993) Role of Helicobacter pylori in the pathogenesis of gastritis, peptic ulcer and gastric cancer. Scand. J. Gastroenterol 28: 3-6. Solnick, J.V. , .Tompkins, L.S. (1993) Helicobacter pylori and gastroduodenal disease: pathogenesis and host-parasite interaction. Infect Ag Dis 1: 294-309.

Stolte, M., Eidt, S. (1993) Healing gastric MALT lymphomas by eradicating H. pylori. Lancet 342: 568. Struyvé, M., Moons, M., Tommassen, J. (1991) Carboxy-terminal phenylalanine is essential for the correct assembly of a bacterial outer membrane protein. J. Mol. Biol. 218: 141-148. Sutcliffe, J.G. (1979) Complete nucleotide sequence of the Eecherichia coli plaemid pBR322. Cold Spring Harb. Symp. Quant Biol. 43: 77-90. Tadayyon, M., Broome-Smith, J.K. (1992) TnblaM -a transposon for directly tagging bacterial genes encoding cell envelope and secreted proteins. Gene 111: 21-26. Taylor, D.N., Blaser, M.J. (1991) The epidemiology of Helicobacter pylori infection. Epidemiol Rev. 13: 42-59. Thomas C.M. (1981) Molecular genetics of broad host range plasmid RK2. Plasmid 5: 10-19. Warren, J.R., Marshall, B. (1983) Unidentified curved bacilli on gastric epithelium in active chronic gastritis. Lancet i: 1273-1275.

LIST OF SEQUENCES (1) GENERAL INFORMATION: (i) APPLICANT: (A) NAME Max-Planck-Gesellschaft zur Foerderung der Wissenschaften e.V. Berlin (B) STREET: Hofgartenstr. 2 (C) CITY: Munich (E) COUNTRY: Germany (F) ZIP CODE: D-80359 (ii) TITLE OF THE INVENTION: Adhesion gene of Helicobacter pylori and polypeptide encoded by the same (iii) NUMBER OF SEQUENCES: 2 (iv) READABLE COMPUTER FORM: (A) TYPE OF MEDIA: Flexible disk (B) COMPUTER: IBM Compatible PC (C) OPERATING SYSTEM: PC-DOS / MS-DOS (D) SOFTWARE OR PROGRAM: Patentln Relay # 1.0 , Version # 1.30 (EPO) (v) CURRENT APPLICATION DETAILS: APPLICATION NUMBER: WO PCT / EP96 / 02545 (2) INFORMATION FOR SEC. FROM IDENT. NO: 1: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 1557 base pairs (B) TYPE: nucleic acid (C) TYPE OF HEBRA: both (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: DNA (genomic) (vi) ORIGINAL SOURCE: (A) ORGANISM: Helicobacter pylori (vii) IMMEDIATE SOURCE: (B) CLONA: alpA (ix) FEATURE: (A) NAME / KEY: CDS (B) PLACE: 1..1554 (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. NO: l: ATG ATA AAA AAG AAT AGA ACG CTG TTT CTT AGT CTA GCC CTT TGC GCT 48 Met lie Lys Asn Arg Thr Leu Phe Leu Ser Leu Ala Leu Cys Ala 1 5 10 15 AGC ATA AGT TAT GCC GAA GAT GAT GGA GGG TTT TTC ACC GTC GGT TAT 96 Ser lie Ser Tyr Ala Glu Asp Asp Gly Gly Phe Phe Thr Val Gly Tyr 20 25 30 CAG CTC GGG CAA GTC ATG CAA GAT GTC CAA AAC CCA GGC GGC GCT AAA 144 Gln Leu Gly Gln Val Met Gln Asp Val Gln Asn Pro Gly Gly Ala Lys 35 40 45 AGC GAC GAA CTC GCC AGA GAG CTT AAC GCT GAT GTA ACG AAC AAC ATT 192 Ser Asp Glu Leu Wing Arg Glu Leu Asn Wing Asp Val Thr Asn Asn lie 50 55 60 TTA AAC AAC AAC ACC GGA GGC AAC ATC GCA GGG GCG TTG AGT AAC GCT 240 Leu Asn Asn Asn Thr Gly Gly Asn lie Wing Gly Wing Leu Ser Asn Wing 65 70 75 80 TTC TCC CAA TAC CTT TAT TCG CTT TTA GGG GCT TAC CCC AA AAA CTC 288 Phe Ser Gln Tyr Leu Tyr Ser Leu Leu Gly Ala Tyr Pro Thr Lys Leu 85 90 95 AAT GGT AGC GAT GTG TCT GCG AAC GCT CTT TTA AGT GGT GCG GTA GGC 336 Asn Gly Ser Asp Val Ser Wing Asn Wing Leu Leu Ser Gly Wing Val Gly 100 105 110 TCT GGG ACT TGT GCG GCT GCA GGG ACG GCT GGT GGC ACT TCT CTT AAC 384 Ser Gly Thr Cys Wing Wing Wing Gly Thr Wing Gly Gly Thr Ser Leu Asn 115 120 125 ACT CAA AGC ACT TGC ACC GTT GCG GGC TAT TAC TGG CTC CCT AGC TTG 432 Thr Gln Ser Thr Cys Thr Val Wing Gly Tyr Tyr Trp Leu Pro Ser Leu 130 135 140 ACT GAC AGG ATT TTA AGC ACG ATC GGC AGC CAG ACT AAC TAC GGC ACG 480 Thr Asp Arg lie Leu Ser Thr lie Gly Ser Gln Thr Asn Tyr Gly Thr 145 150 155 160 AAC ACC AAT TTC CCC AAC ATG CAA CAA CAA CAG CTC ACC TAC TTG AAT GCG 528 Asn Thr Asn Phe Pro Asn Met Gln Gln Gln Leu Thr Tyr Leu Asn Wing 165 170 175 GGG AAT GTG TTT TTT AAT GCG ATG AAT AAG GCT TTA GAG AAT AAG AAT 576 Gly Asn Val Phe Phe Asn Wing Met Asn Lys Wing Leu Glu Asn Lys Asn 180 185 190 GGA ACT AGT AGT GCT AGT GGA ACT AGT GGT GCG ACT GGT TCA GAT GGT 624 Gly Thr Ser Ser Wing Ser Gly Thr Ser Gly Wing Thr Gly Ser Asp Gly 195 200 205 CAA ACT TAC TCC ACÁ CAA GCT ATC CAA TAC CTT CAA GGC CAA CAA AAT 672 Gln Thr Tyr Ser Thr Gln Ala lie Gln Tyr Leu Gln Gly Gln Gln Asn 210 215 220 ATC TTA AAT AAC GCA GCG AAC TTG CTC AAG CAA GAT GAA TTG CTC TTA 720 lie Leu Asn Asn Wing Wing Asn Leu Leu Lys Gln Asp Glu Leu Leu Leu 225 230 235 240 GAA GCT TTC AAC TCT GCC GTA GCC GCC AAC ATT GGG AAT AAG GAA TTC 768 Glu Ala Phe Asn Ser Ala Ala Ala Ala Asn lie Gly Asn Lys Glu Phe 245 250 255 AAT TCA GCC GCT TTT ACA GGT TTG GTG CAA GGC ATT ATT GAT CAA TCT 816 Asn Ser Wing wing Phe Thr Gly Leu Val Gln Gly lie lie Asp Gln Ser 260 265 270 CAA GCG GTT TAT AAC GAG CTC ACT AAA AAC ACC ATT AGC GGG AGT GCG 864 Gln Wing Val Tyr Asn Glu Leu Thr Lys Asn Thr lie Ser Gly Ser Wing 275 280 285 GTT ATT AGC GCT GGG ATA AAC TCC AAC CAA GCT AAC GCT GTG CAA GGG 912 Val lie Ser Wing Gly lie Asn Ser Asn Gln Wing Asn Wing Val Gln Gly 290 295 300 CGC GCT AGT CAG CTC CCT AAC GCT CTT TAT AAC GCG CAA GTA ACT TTG 960 Arg Ala Ser Gln Leu Pro Asn Ala Leu Tyr Asn Ala Gln Val Thr Leu 305 310 315 320 GAT AAA ATC AAT GCG CTC AAT AAT CAA GTG AGA AGC ATG CCT TAC TTG 1008 Asp Lys lie Asn Ala Leu Asn Asn Gln Val Ara Ser Met Pro Tyr Leu 325 330 335 CCC CAA TTC AGA GCC GGG AAC AGC CGT TCA ACG AAT ATT TTA AAC GGG 1056 Pro Gln Phe Arg Wing Gly Asn Ser Arg Ser Thr Asn lie Leu Asn Gly 340 345 350 TTT AC ACC AAA ATA GGC TAT AAG CAA TTC TTC GGG AAG AAA AGG AAT 1104 Phe Tyr Thr Lys lie Gly Tyr Lys Gln Phe Phe Gly Lys Lys Arg Asn 355 360 365 ATC GGT TTG CGC TAT TAT GGT TTC TTT TCT TAT AAC GGA GCG AGC GTG 1152 lie Gly Leu Arg Tyr Tyr Gly Phe Phe Ser Tyr Asn Gly Wing Ser Val 370 375 380 GGC TTT AGA TCC ACT CAA AAT AAT GTA GGG TTA TAC ACT TAT GGG GTG 1200 Gly Phe Arg Ser Thr Gln Asn Asn Val Gly Leu Tyr Thr Tyr Gly Val 385 390 395 400 GGG ACT GAT GTG TTG TAT AAC ATC TTT AGC CGC TCC TAT CAA AAC CGC 1248 Gly Thr Asp Val Leu Tyr Asn lie Phe Ser Arg Ser Tyr Gln Asn Arg 405 410 415 TCT GTG GAT ATG GGC TTT TTT AGC GGT ATC CAA TTA GCC GGT GAG ACC 1296 Ser Val Asp Met Gly Phe Phe Ser Gly lie Gln Leu Ala Gly Glu Thr 420 425 430 TTC CAA TCC ACG CTC AGA GAT GAC CCC AAT GTG AAA TTG CAT GGG AAA 1344 Phe Gln Ser Thr Leu Arg Asp Asp Pro Asn Val Lys Leu His Gly Lys 435 440 445 ATC AAT AAC ACG CAC TTC CAG TTC CTC TTT GAC TTC GGT ATG AGG ATG 1392 lie Asn Asn Thr His Phe Gln Phe Leu Phe Asp Phe Gly Met Arg Met 450 455 460 AAC TTC GGT AAG TTG GAC GGG AAA TCC AAC CGC CAC AAC CAG CAC ACG 1440 Asn Phe Gly Lys Leu Asp Gly Lys Ser Asn Arg His Asn Gln His Thr 465 470 475 480 GTG GAA TTT GGC GTA GTG GTG CCT ACG ATT TAT AAC ACT TAT TAC AAA 1488 Val Glu Phe Gly Val Val Val Pro Thr lie Tyr Asn Thr Tyr Tyr Lys 485 490 495 TCA GCA GGG ACT ACC GTG AAG TAT TTC CGT CCT TAT AGC GTT TAT TGG 1536 Ser Wing Gly Thr Thr Val Lys Tyr Phe Arg Pro Tyr Ser Val Tyr Trp 500 505 510 TCT TAT GGG TAT TCA TTC TAA 1557 Ser Tyr Gly Tyr Ser Phe 515 (2) INFORMATION FOR SEC. FROM IDENT. NO: 2: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 518 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: protein (xi) DESCRIPTION OF THE SEQUENCE: SEC. FROM IDENT. N0: 2: Met lie Lys Lys Asn Arg Thr Leu Phe Leu Ser Leu Ala Leu Cys Wing 1 5 10 15 Ser lie Tyr Wing Glu Asp Asp Gly Gly Phe Phe Thr Val Gly Tyr 20 25 30 Gln Leu Gly Gln Val Met Gln Asp Val Gln Asn Pro Gly Gly Ala Lys 35 40 45 Being Asp Glu Leu Wing Arg Glu Leu Asn Wing Asp Val Thr Asn Asn lie 50 55 60 Leu Asn Asn Asn Thr Gly Gly Asn lie Wing Gly Wing Leu Ser Asn Wing 65 70 75 80 Phe Ser Gln Tyr Leu Tyr Ser Leu Leu Gly Wing Tyr Pro Thr Lys Leu 85 90 95 Asn Gly Ser Asp Val Ser Wing Asn Wing Leu Leu Ser Gly Wing Val Gly 100 105 110 Be Gly Thr Cys Wing Wing Wing Gly Thr Wing Gly Gly Thr Ser Leu Asn 115 120 125 Thr Gln Ser Thr Cys Thr Val Wing Gly Tyr Tyr Trp Leu Pro Ser Leu 130 135 140 Thr Asp Arg lie Leu Ser Thr lie Gly Ser Gln Thr Asn Tyr Gly Thr 145 150 155 160 Aen Thr Asn Phe Pro Asn Met Gln Gln Gln Leu Thr Tyr Leu Asn Wing 165 170 175 Gly Aen Val Phe Phe Aen Wing Met Aen Lye Wing Leu Glu Asn Lys Asn 180 185 190 Gly Thr Ser Ser Wing Ser Gly Thr Ser Gly Wing Thr Gly Ser Asp Gly 195 200 205 Gln Thr Tyr Ser Thr Gln Ala lie Gln Tyr Leu Gln Gly Gln Gln Asn 210 215 220 lie Leu Asn Asn Ala Ala Asn Leu Leu Lys Gln Asp Glu Leu Leu Leu 225 230 235 240 Glu Ala Phe Asn Ser Ala Ala Ala Ala Ala Asn lie Gly Asn Lys Glu Phe 245 250 255 Asn Be Wing Wing Phe Thr Gly Leu Val Gln Gly lie lie Asp Gln Ser 260 265 270 Gln Ala Val Tyr Asn Glu Leu Thr Lys Asn Thr lie Ser Gly Ser Wing 275 280 285 Val lie Be Wing Gly lie Asn Be Asn Gln Wing Asn Wing Val Gln Gly 290 295 300 Arg Ala Ser Gln Leu Pro Asn Ala Leu Tyr Asn Ala Gln Val Thr Leu 305 310 315 320 Asp Lys lie Asn Ala Leu Asn Asn Gln Val Arg Ser Met Pro Tyr Leu 325 330 335 Pro Gln Phe Arg Wing Gly Asn Ser Arg Ser Thr Asn lie Leu Asn Gly 340 345 350 Phe Tyr Thr Lys lie Gly Tyr Lys Gln Phe Phe Gly Lye Lys Arg Asn 355 360 365 lie Gly Leu Arg Tyr Tyr Gly Phe Phe Ser Tyr Asn Gly Wing Ser Val 370 375 380 Gly Phe Arg Ser Thr Gln Asn Asn Val Gly Leu Tyr Thr Tyr Gly Val 385 390 395 400 Gly Thr Asp Val Leu Tyr Asn lie Phe Ser Arg Ser Tyr Gln Asn Arg 405 410 415 Ser Val Asp Met Gly Phe Phe Ser Gly lie Gln Leu Ala Gly Glu Thr 420 425 430 Phe Gln Ser Thr Leu Arg Asp Asp Pro Asn Val Lys Leu His Gly Lys 435 440 445 lie Asn Asn Thr His Phe Gln Phe Leu Phe Asp Phe Gly Met Arg Met 450 455 460 Asn Phe Gly Lys Leu Asp Gly Lys Ser Asn Arg His Asn Gln His Thr 465 470 475 480 Val Glu Phe Gly Val Val Val Pro Thr lie Tyr Asn Thr Tyr Tyr Lys 485 490 495 Be Ala Gly Thr Thr Val Lys Tyr Phe Arg Pro Tyr Ser Val Tyr Trp 500 505 510 Ser Tyr Gly Tyr Ser Phe 515 It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is the conventional one for the manufacture of the objects to which it relates. Having described the invention as above, property is claimed as contained in the following:

Claims (16)

1. A DNA molecule, characterized in that it comprises: (a) the nucleotide sequence shown in SEC. FROM IDENT. N0: 1; (b) a nucleotide sequence which corresponds to the sequence according to (a) within the scope of the degeneracy of the genetic code, or (c) a nucleotide sequence which hybridizes to the sequences according to (a) or / and (b) under conditions of restriction.

2. The DNA molecule according to claim 1, characterized in that at the nucleotide level it exides a homology of at least 80% with the nucleotide sequence shown in SEQ. FROM IDENT. NO: l.

3. The DNA molecule according to claim 1 or 2, characterized in that it has a length of at least 15 nucleotides.

4. The DNA molecule according to one of claims 1 to 3, characterized in that it encodes a polypeptide with the ability to adhere to human cells.

5. A vector, characterized in that it contains at least one copy of the DNA molecule according to one of claims 1 to 4.

6. A cell, characterized in that it is transformed with a vector as claimed in accordance with claim 5.

7. A polypeptide, characterized in that it is encoded by a DNA molecule according to one of claims 1 to 4.

8. The polypeptide according to claim 7, characterized in that it comprises: (a) the amino acid sequence shown in SEQ. FROM IDENT. NO: 2; or (b) an amino acid sequence which immunologically cross-reacts with the sequence of compliance with (a).

9. A polypeptide, characterized in that it is capable of adhering to human cells.

10. A process for the production of a polypeptide according to one of claims 7 to 9, characterized in that a cell is transformed with a DNA molecule according to one of claims 1 to 4, or a vector according to claim 5 , the transformed cell is cultured under conditions in which the expression of the polypeptide takes place and the polypeptide is isolated from the cell and / or the culture supernatant.

11. The use of a polypeptide according to one of claims 7 to 9, characterized in that it is used as an immunogen to produce antibodies.

12. An antibody against a polypeptide, characterized in that it is according to one of claims 7 to 9.

13. The antibody according to claim 12, characterized in that it is directed against the N-terminal part of the amino acid sequence shown in SEQ. FROM IDENT. NO: 2

14. A pharmaceutical composition, characterized in that it contains a DNA molecule according to one of claims 1 to 4, a polypeptide according to one of claims 7 to 9 or an antibody according to claims 12 or 13, as the active substance, optionally together with pharmaceutically common auxiliaries, diluents, additives and carriers.

15. The use of a pharmaceutical composition according to claim 14, characterized in that it is for the diagnosis of a Helicobacter pylori infection.

16. The use of a pharmaceutical composition according to claim 14, characterized in that it is for the production of an agent for the prevention or treatment of an infection by Helicobacter pylori.