CA2341637A1 - Chlamydia antigens and corresponding dna fragments and uses thereof - Google Patents

Abstract

In summary of this disclosure, the present invention provides a method of nucleic acid, including DNA, immunization of a host, including humans, again st disease caused by infection by a strain of Chlamydia, specifically C. pneumoniae, employing a vector, containing a nucleotide sequence encoding an y of the following polypeptides: CPN 100111, CPN 100224, CPN 100230, CPN 10023 1, CPN 100232, CPN 100235, CPN 100394, CPN 100395 of a strain of Chlamydia pneumoniae and a promoter to effect expression of any of the polypeptides in the host. Modifications are possible within the scope of this invention.</SD OAB>

Description

CHLAMYDIA ANTIGENS AND CORRESPONDING DNA FRAGMENTS
AND USES THEREOF
RELATED U.S. APPLICATION
The present patent application claims priority to the following United States provisional patent applications: U.S.S.Ns. 60/097,187, 60/097,188, 60/097,189, 60/097,190, 60/097,195, 60/097,196, and 60/097,197, filed August 20, 1998, and U.S.S.N. 60/097,191, filed August 27, 1998.
FIELD OF THE INVENTION
The present invention relates to Chlamydia antigens and corresponding DNA
molecules, 1 o which can be used in methods to prevent and treat disease caused by Chlamydia infection in mammals, such as humans.
BACKGROUND OF THE INVENTION
Chlamydiae are prokaryotes. They exhibit morphologic and structural similarities to Gram negative bacteria including a trilaminar outer membrane, which contains lipopolysaccharide and 15 several membrane proteins. Chlamydiae are differentiated from other bacteria by their morphology and by a unique developmental cycle. They are obligate intracellular parasites with a unique biphasic life cycle consisting of a metabolically inactive but infectious extracellular stage and a replicating but non-infectious intracellular stage. The replicative stage of the life-cycle takes place within a membrane-bound inclusion which sequesters the bacteria away from the cytoplasm of the 2o infected host cell.
Because chlamydiae are small and multiply only within susceptible cells they were long thought to be viruses. However, they have many characteristics in common with other bacteria:
(1) they contain both DNA and RNA, (2) they divide by binary fission, (3) their cell envelopes resemble those of other Gram-negative bacteria, (4) they contain ribosomes similar to those of 25 other bacteria, and (5) they are susceptible to various antibiotics.
Chlamydiae can be seen in the light microscope, and the genome is about one-third the size of the Escherichia toll genome.
Many different strains of chlamydiae have been isolated from birds, man, and other mammals, and these strains can be distinguished on the basis of host range, virulence, _1_ CONFIRMATION COPT

pathogenesis, and antigenic composition. There is strong homology of DNA
within each species, but surprisingly little between species, suggesting long-standing evolutionary separation.
C. trachomatis has a high degree of host specificity, being almost completely limited to man; it causes ocular and genitourinary infections of widely varying severity.
In contrast, C. psittaci strains are rare in man but are found in a wide range of birds and also in wild, domestic, and laboratory mammals, where they multiply in cells of many organs.
C. pneumoniae is a common human pathogen, originally described as the TWAR
strain of C. psittaci, but subsequently recognized to be a new species. C. pneumoniae is antigenically, genetically, and morphologically distinct from other Chlamydia species (C.
trachomatis, 1 o C. pecorum and C. psittaci). It shows 10% or less DNA sequence homology with either of C. trachomatis or C. psittaci and so far appears to consist of only a single strain, TWAR.
C. pneumoniae is a common cause of community acquired pneumonia, less frequent only than Streptococcus pneumoniae and Mycoplasma pneumoniae. Grayston et al., J.
Infect. Dis. 168:
1231 (1995); Campos et al., Invest. Ophthalmol. Vis. Sci. 36: 1477 (1995), each incorporated herein by reference. It can also cause upper respiratory tract symptoms and disease, including bronchitis and sinusitis. See, e.g., Grayston et al., J. Infect. Dis. 168:
1231 (1995); Campos et al., Invest. Ophthalmol. Vis. Sci. 36: 1477 (1995); Grayston et al., J. Infect.
Dis. 161: 618 (1990);
Marrie, Clin. Infect. Dis. 18: 501 (1993). The great majority of the adult population (over 60%) has antibodies to C. pneumoniae (Wang et al., Chlamydial Infections, Cambridge University Press, 2o Cambridge, p. 329 (1986)), indicating past infection which was unrecognized or asymptomatic.
C. pneumoniae infection usually presents as an acute respiratory disease (i.e., cough, sore throat, hoarseness, and fever; abnormal chest sounds on auscultation). For most patients, the cough persists for 2 to 6 weeks, and recovery is slow. In approximately 10% of these cases, upper respiratory tract infection is followed by bronchitis or pneumonia.
Furthermore, during a C. pneumoniae epidemic, subsequent co-infection with pneumococcus has been noted in about half of these pneumonia patients, particularly in the infirm and the elderly. As noted above, there is more and more evidence that C. pneumoniae infection is also linked to diseases other than respiratory infections.
The reservoir for the organism is presumably people. In contrast to C.
psittaci infections, 3o there is no known bird or animal reservoir. Transmission has not been clearly defined. It may result from direct contact with secretions, from formites, or from airborne spread. There is a long incubation period, which may last for many months. Based on analysis of epidemics, C. pneumoniae appears to spread slowly through a population (case-to-case interval averaging 30 days) because infected persons are inefficient transmitters of the organism. Susceptibility to C. pneumoniae is universal. Reinfections occur during adulthood, following the primary infection as a child. C. pneumoniae appears to be an endemic disease throughout the world, noteworthy for superimposed intervals of increased incidence (epidemics) that persist for 2 to 3 years. C.
trachomatis infection does not confer cross-immunity to C. pneumoniae.
Infections are easily treated with oral antibiotics, tetracycline or erythromycin (2 g/day, for at least 10 to 14 days). A
to recently developed drug, azithromycin, is highly effective as a single-dose therapy against chlamydial infections.
In most instances, C.'. pneumoniae infection is mild and without complications, and up to 90% of infections are subacute or unrecognized. Among children in industrialized countries, infections have been thought to be rare up to the age of five years, although a recent study has 15 reported that many children in this age group show PCR evidence of infection despite being seronegative, and estimates a prevalence of 17-19% in 2-4 years old. See, Normann et al., Acta Paediatrica, 87: 23-27 ( 1998). In developing countries, the seroprevalence of C. pneumoniae antibodies among young children is elevated, and there are suspicions that C.
pneumoniae may be an important cause of acute lower respiratory tract disease and mortality for infants and children in 2o tropical regions of the world.
From seroprevalence studies and studies of local epidemics, the initial C.
pneumoniae infection usually happens between the ages of 5 and 20 years. In the USA, fox example, there are estimated to be 30,000 cases of childhood pneumonia each year caused by C.
pneumoniae.
Infections may cluster among groups of children or young adults (e.g., school pupils or military 25 conscripts).
C. pneumoniae causes 10 to 25% of community-acquired lower respiratory tract infections (as reported from Sweden, Italy, Finland, and the USA). During an epidemic, C.
pneumonia infection may account for 50 to 60% of the cases of pneumonia. During these periods, also, more episodes of mixed infections with S. pneumoniae have been reported.

Reinfection during adulthood is common; the clinical presentation tends to be milder.
Based on population seroprevalence studies, there tends to be increased exposure with age, which is particularly evident among men. Some investigators have speculated that a persistent, asymptomatic C. pneumoniae infection state is common.
In adults of middle age or older, C. pneumoniae infection may progress to chronic bronchitis and sinusitis. A study in the USA revealed that the incidence of pneumonia caused by C. pneumoniae in persons younger than 60 years is 1 case per 1,000 persons per year; but in the elderly, the disease incidence rose three-fold. C pneumoniae infection rarely leads to hospitalization, except in patients with an underlying illness.
to Of considerable importance is the association of atherosclerosis and C.
pneumoniae infection. There are several epidemiological studies showing a correlation of previous infections with C. pneumoniae and heart attacks, coronary artery and carotid artery disease. See, Saikku et al., Lancet 2: 983 (1988); Thom et al., JAMA 268: 68 (1992); Linnanmaki et al., Circulation 87:
1030 (1993); Saikku et al., Annals Int. Med. 116: 273 (1992); Melnick et al., Am. J. Med. 95: 499 i 5 ( 1993). Moreover, the organisms has been detected in atheromas and fatty streaks of the coronary, carotid, peripheral arteries and aorta. See, Shor et al., South African Med.
J. 82: 158 (1992); Kuo et al., J. Infect. Dis. 167: 841 (i993); Kuo et al., Arteriosclerosis and Thrombosis 13: 1500 (1993);
Campbell et al., J. Infect. Dis. 172: 585 (1995); Chiu et al., Circulation 96:
2144-2148 (1997).
Viable C. pneumoniae has been recovered from the coronary and carotid artery.
Ramirez et al., 2o Annals Int. Med. 125: 979 (1996); Jackson et al., Abst. K121, p272, 36th ICAAC, New Orleans ( 1996). Furthermore, it has been shown that C. pneumoniae can induce changes of atherosclerosis in a rabbit model. See, Fong et al., (1997) Journal of Clinical Microbiololo~
35: 48. Taken together, these results indicate that it is highly probable that C. pneumoniae can cause atherosclerosis in humans, though the epidemiological importance of chlamydial atherosclerosis 25 remains to be demonstrated.
A number of recent studies have also indicated an association between C.
pneumoniae infection and asthma. Infection has been linked to wheezing, asthmatic bronchitis, adult-onset asthma and acute exacerbation of asthma in adults, and small-scale studies have shown that prolonged antibiotic treatment was effective at greatly reducing the severity of the disease in some 3o individuals. Hahn et al., Ann Allergy Asthma Immunol. 80: 45-49 (1998);
Hahn et al., Epidemiol Infect. 117: 513-517 (1996); Bjornsson et al., Scand J Infect. Dis. 28: 63-69 (1996); Hahn, J. Fam.
Pract. 41: 345-351 (1995); Allegra et al., Eur. Respir. J. 7: 2165-2168 (1994); Hahn et al., JAMA
266: 225-230 (1991).
In light of these results, a protective vaccine against disease caused by C.
pneumoniae s infection would be of considerable importance. There is not yet an effective vaccine for human C. pneumoniae infection. Nevertheless, studies with C. trachomatis and G
psittaci indicate that this is an attainable goal. For example, mice which have recovered from a lung infection with C. trachomatis are protected from infertility induced by a subsequent vaginal challenge. Pal et al., Infection and Immunity 64: 5341 (1996). Similarly, sheep immunized with inactivated C. psittaci t0 were protected from subsequent chlamydial-induced abortions and stillbirths. Jones et al., Iraccine 13: 715 ( 1995). Protection from chlamydial infections has been associated with Th 1 immune responses, particularly the induction of INFy-producing CD4+ T cells.
Igietsemes et al., Immunology 5: 317 (1993). The adoptive transfer of CD4+ cell lines or clones to nude or SCID
mice conferred protection from challenge or cleared chronic disease (Igietseme et al., Regional ~5 Immunology S: 317 (1993); Magee et al., Regional Immunology 5: 305 (1993)), and in vivo depletion of CD4+ T cells exacerbated disease post-challenge (Larders et al., Infection 8c Immunity 59: 3774 (1991); Magee et al., Infection ~c Immunity 63: 516 (1995)).
However, the presence of sufficiently high titres of neutralizing antibody at mucosal surfaces can also exert a protective effect. Cotter et al., Infection and Immunity 63: 4704 (1995).
2o The extent of antigenic variation within the species C. pneumoniae is not well characterized. Serovars of C. trachomatis are defined on the basis of antigenic variation in major outer membrane proteins (MOMP), but published C. pneumoniae MOMP gene sequences show no variation between several diverse isolates of the organism. See, Campbell et al., Infection and Immunity 58: 93 (1990); McCafferty et al., Infection and Immunity 63: 2387-9 (1995); Knudsen et 2s al., Third Meeting of the European Society for Chlamydia Research, Vienna (1996). Regions of the protein known to be conserved in other chlamydial MOMPs are conserved in C. pneumoniae.
See, Campbell et al., Infection and Immunity 58: 93 (1990); McCafferty et al., Infection and Immunity 63: 2387-9 (1995). One study has described a strain of C. pneumoniae with a MOMP of greater that usual molecular weight, but the gene for this has not been sequenced. Grayston et al., 3o J. Infect. Dis. 168: 1231 (1995). Partial sequences of outer membrane protein 2 from nine diverse isolates were also found to be invariant. Ramirez et al., Annals Int. Med.
125: 979 (1996). The -s-WO 00/111$3 PCT/IB99/01449 genes for HSP60 and HSP70 show little variation from other chlamydial species, as would be expected. The gene encoding a 76 kDa antigen has been cloned from a single strain of C. pneumoniae. It has no significant similarity with other known chlamydial genes. Marrie, Clin.
Infect. Dis. 18: 501 (1993).
Many antigens recognized by immune sera to C. pneumoniae are conserved across all chlamydiae, but 98kDa, 76 kDa and S4 kDa proteins may be C. pneumoniae-specific. Campos et al., Invest. Ophthalmol. Vis. Sci. 36: 1477 (1995); Marrie, Clin. Infect. Dis.
18: 501 (1993);
Wiedmann-AI-Ahmad et al., Clin. Diagn. Lab. Immunol. 4: 700-704 (1997).
Immunoblotting of isolates with sera from patients does show variation of blotting patterns between isolates, 1o indicating that serotypes C. pneumoniae may exist. Grayston et al., J.
Infect. Dis. 168: 1231 (1995); Ramirez et al., Annals Int. Med. 125: 979 (1996). However, the results are potentially confounded by the infection status of the patients, since immunoblot profiles of a patient's sera change with time post-infection. An assessment of the number and relative frequency of any serotypes, and the defining antigens, is not yet possible.
Thus, a need remains for effective compositions for preventing, treating, and diagnosing Chlamydia infections.
SUMMARY OF THE INVENTION
In one aspect, the present invention provides purified and isolated DNA
molecules, which encode Chlamydia that can be used in methods to prevent, treat, and diagnose Chlamydia infection. The present invention provides eight separate preferred DNA
molecules, each individually and separately defined by one SEQ ID NOS: 1, 3, S, 7, 9, 11, 13, or 15. Further provided in the present invention are eight separate preferred polypeptides, each individually and separately defined by one of SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, or 16. Those skilled in the art will appreciate that the invention also includes DNA molecules that encode mutants, variants, and derivatives of such polypeptides, which result from the addition, deletion, or substitution of non-essential amino acids as described herein. The invention also includes RNA
molecules corresponding to the DNA molecules of the invention.
In addition to the DNA and RNA molecules, the invention includes the corresponding polypeptides and monospecific antibodies that specifically bind to such polypeptides.

The present invention has wide application and includes expression cassettes, vectors, and cells transformed or transfected with the polynucleotides of the invention.
Accordingly, the present invention provides: (i) a method for producing a polypeptide of the invention in a recombinant host system and related expression cassettes, vectors, and transformed or transfected cells; (ii) a live vaccine vectors such as viral or bacterial live vaccine vectors, including, pox virus, alphavirus, Salmonella typhimurium, or Vibrio cholerae vector, containing a polynucleotide of the invention, such vaccine vectors being useful for, e.g., preventing and treating Chlamydia infection, in combination with a diluent or carrier, and related pharmaceutical compositions and associated therapeutic and/or prophylactic methods; (iii) a therapeutic and/or prophylactic method involving administration of an RNA or DNA molecule of the invention, either in a naked form or formulated with a delivery vehicle, a polypeptide or combination of polypeptides, or a monospecific antibody of the invention, and related pharmaceutical compositions; (iv) a method for diagnosing the presence of Chlamydia in a biological sample, which can involve the use of a DNA or RNA
molecule, a monospecific antibody, or a polypeptide of the invention; and (v) a method for purifying a polypeptide of the invention by antibody-based affinity chromatography.
BRIEF DESCRIPTION OF THE DRAWINGS
The present invention will be fuxther understood from the following description with reference to the drawings, in which:
FIG. 1 shows the nucleotide sequence (top sequence) and the deduced amino acid sequence of the full-length (middle sequence) and processed (bottom sequence) CPN
100111 polypeptide from Chlamydia pneumoniae.
FIG. 2 shows the restriction enzyme analysis of the nucleotide sequence encoding C. pneumoniae CPN100111.
FIG. 3 shows the nucleotide sequence (top sequence) and the deduced amino acid sequence of the full-length (middle sequence) and processed (bottom sequence) CPN
100224 polypeptide from Chlamydia pneumoniae.
FIG. 4 shows the restriction enzyme analysis of the nucleotide sequence encoding the C. pneumoniae CPN 100224.

FIG. 5 shows the nucleotide sequence (top sequence) and the deduced amino acid sequence (bottom sequence) of the CPN100230 polypeptide from Chlamydia pneumoniae.
FIG. 6 shows the restriction enzyme analysis of the nucleotide sequence encoding the C. pneumoniae CPN100230.
FIG. 7 shows the nucleotide sequence (top sequence) and the deduced amino acid sequence (bottom sequence) of the CPN100231 polypeptide from Chlamydia pneumoniae.
FIG. 8 shows the restriction enzyme analysis of the nucleotide sequence encoding the C. pneumoniae CPN 100231.
FIG. 9 shows the nucleotide sequence (top sequence) and the deduced amino acid sequence (bottom sequence) of the CPN100232 polypeptide from Chlamydia pneumoniae.
FIG. 10 shows the restriction enzyme analysis of the nucleotide sequence encoding the C. pneumoniae CPN 100232.
FIG. 11 shows the nucleotide sequence (top sequence) and the deduced amino acid sequence (bottom sequence) of the CPN100233 polypeptide from Chlamydia pneumoniae.
15 FIG. 12 shows the restriction enzyme analysis of the nucleotide sequence encoding the C. pneumoniae CPN100233.
FIG. 1'3 shows the nucleotide sequence (top sequence) and the deduced amino acid sequence of the full-length (middle sequence) and processed (bottom sequence) polypeptide from Chlamydia pneumoniae.
2o FIG. 14 shows the restriction enzyme analysis of the nucleotide sequence encoding the C. pneumoniae CPN100394.
FIG. 1 S shows the nucleotide sequence (top sequence) and the deduced amino acid sequence (bottom sequence) of the CPN100395 polypeptide from Chlamydia pneumoniae.
FIG. 16 shows the restriction enzyme analysis of the nucleotide sequence encoding the 25 C. pneumoniae CPN 100395.
_g_ DETAILED DESCRIPTION OF THE INVENTION
In the C. pneumoniae genome, open reading frames (ORFs) encoding chlamydial polypeptides have been identified. These polypeptides include polypeptides permanently found in the bacterial membrane structure, polypeptides that are present in the external vicinity of the bacterial membrane, polypeptides permanently found in the inclusion membrane structure, polypeptides that are present in the external vicinity of the inclusion membrane, and polypeptides that are released into the cytoplasm of the infected cell. These polypeptides can be used in vaccination methods for preventing and treating Chlamydia infection.
According to a first aspect of the invention, there are provided isolated polynucleotides encoding the precursor and mature forms of Chlamydia polypeptides.
An isolated polynucleotide of the invention encodes a polypeptide having an amino acid sequence that is homologous to a Chlamydia amino acid sequence, the Chlamydia amino acid sequence being selected from the group consisting of the amino acid sequences as shown in SEQ
ID NOS: 2, 4, 6, 8, 10, 12, 14, or 16.
The term "isolated polynucleotide" is defined as a polynucleotide removed from the environment in which it naturally occurs. For example, a naturally-occurring DNA molecule present in the genome of the bacteria is not isolated, but the same molecule separated from the remaining part of the bacterial genome, as a result of, e.g., a cloning event (amplification), is isolated. Typically, an isolated DNA molecule is free from DNA regions (e.g., coding regions) 2o with which it is immediately contiguous at the S' or 3' end, in the naturally occurring genome.
Such isolated polynucleotides could be part of a vector or a composition and still be isolated in that such a vector or composition is not part of its natural environment.
A polynucleotide of the invention can be in the form of RNA or DNA (e.g., cDNA, genomic DNA, or synthetic DNA), or modifications or combinations thereof. The DNA can be double-stranded or single-stranded, and, if single-stranded, can be the coding strand or the non-coding (anti-sense) strand. The sequence that encodes a polypeptide of the invention as shown in SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14 or 16 can be: (a) the coding sequence as shown in either SEQ ID NOS: 1, 3, S, 7, 9, 11, 13 and 15; or (b) a ribonucleotide sequence derived by transcription of (a); or (c) a different coding sequence; this latter, as a result of the redundancy or degeneracy of the genetic code, encodes the same polypeptides as the DNA
molecules of which the nucleotide sequences are illustrated in SEQ ID NOS: l, 3, 5, 7, 9, 11, 13, and 15..
By "homologous amino acid sequence" is meant an amino acid sequence that differs from an amino acid sequence shown in SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14 or 16, only by one or more conservative amino acid substitutions, or by one or more non-conservative amino acid substitutions, deletions, or additions located at positions at which they do not destroy the specific antigenicity of the polypeptide.
Preferably, such a sequence is at least 75%, more preferably 80%, and most preferably 90%
identical to an amino acid sequence shown in SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14 or 16.
l0 Homologous amino acid sequences include sequences that are identical or substantially identical to an amino acid sequence as shown in SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14 and 16. By "amino acid sequence substantially identical" is meant a sequence that is at least 90%, preferably 95%, more preferably 97%, and most preferably 99% identical to an amino acid sequence of reference and that preferably differs from the sequence of reference, if at all, by a majority of t 5 conservative amino acid substitutions.
Conservative amino acid substitutions typically include substitutions among amino acids of the same class. These classes include, for example, (a) amino acids having uncharged polar side chains, such as asparagine, glutamine, serine, threonine, and tyrosine; (b) amino acids having basic side chains, such as lysine, arginine, and histidine; (c) amino acids having acidic side chains, such 20 as aspartic acid and glutamic acid; and (d) amino acids having nonpolar side chains, such as glycine, alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan, and cysteine.
Homology is typically measured using sequence analysis software (e.g., Sequence Analysis Software Package of the Genetics Computer Group, University of Wisconsin Biotechnology 25 Center, 1710 University Avenue, Madison, WI 53705). Similar amino acid sequences are aligned to obtain the maximum degree of homology (i. e., identity). To this end, it may be necessary to introduce gaps into the sequence. Once the optimal alignment has been set up, the degree of homology (i.e., identity) is established by recording all of the positions in which the amino acids of both sequences are identical, relative to the total number of positions.

Alternatively, homology can be determined by aligning the candidate sequence and the reference sequence using an alignment tool, such as the dynamic programming algorithm described in Needleman et al., J. Mol. Blol. 48: 443 -( 1970), and the Align Program, a commercial software package produced by DNAstar, Inc., the teachings of which are incorporated by reference herein. After the initial alignment is made, it can be refined by comparison to a multiple sequence alignment of a family of related proteins. Once the alignment between the candidate and reference sequences is made and refined, a percent homology score is calculated. The individual amino acids of each sequence are compared sequentially according to their similarity to each other.
Similarity factors include similar size, shape and electrical charge. One particularly 1 o preferred method of determining amino acid similarities is the PAM250 matrix described in Dayhoff et al., 5 ATLAS OF PROTEIN SEQUENCE AND STRUCTURE 345-352 (1978 &
Supp.), incorporated by reference herein. A similarity score is first calculated as the sum of the aligned pairwise amino acid similarity scores. Insertions and deletions are ignored for the purposes of percent homology and identity. Accordingly, gap penalties are not used in this calculation. The 15 raw score is then normalized by dividing it by the geometric mean of the scores of the candidate compound and the reference sequence. The geometric mean is the square root of the product of these scores. The normalized raw score is the percent homology.
Preferably, a homologous sequence is one that is at least 45%, more preferably 60%, and most preferably 85% identical to a coding sequence of SEQ ID NOS: 1, 3, 5, 7, 9, 11, 13, and 15.
2o Polypeptides having a sequence homologous to one of the sequences shown in SEQ ID
NOS: 2, 4, 6, 8, 10, 12, 14 or 16 include naturally-occurring allelic variants, as well as mutants and variants or any other non-naturally-occurring variants that are analogous in terms of antigenicity, to a polypeptide having a sequence as shown in SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14 or 16.
An allelic variant is an alternate form of a polypeptide that is characterized as having a 25 substitution, deletion, or addition of one or more amino acids that does not substantially alter the biological function of the polypeptide. By "biological function" is meant the function of the polypeptide in the cells in which it naturally occurs, even-if the function is not necessary for the growth or survival of the cells. For example, the biological function of a porin is to allow the entry into cells of compounds present in the extracellular medium. The biological function is distinct 3o from the antigenic function. A polypeptide can have more than one biological function.

Allelic variants are very common in nature. For example, a bacterial species, e.g., C. pneumoniae, is usually represented by a variety of strains that differ from each other by minor allelic variations. Indeed, a polypeptide that fulfills the same biological function in different strains can have an amino acid sequence that is not identical in each of the strains. Such an allelic variation may be equally reflected at the polynucleotide level.
Support for the use of allelic variants of polypeptide antigens comes from, e.g., studies of the Chlamydial MOMP antigen. The amino acid sequence of the MOMP varies from strain to strain, yet cross-strain antibody binding plus neutralization of infectivity occurs, indicating that the MOMP, when used as an immunogen, is tolerant of amino acid variations.
to Polynucleotides, e.g., DNA molecules, encoding allelic variants can easily be retrieved by polymerise chain reaction (PCR) amplification of genomic bacterial DNA
extracted by conventional methods. This involves the use of synthetic oligonucleotide primers matching upstream and downstream of the 5' and 3' ends of the encoding domain. Suitable primers can be designed according to the nucleotide sequence information provided in SEQ ID
NOS: 1, 3, 5, 7, 9, 15 11, 13 and 15. Typically, a primer can consist of 10 to 40, preferably 15 to 25 nucleotides. It may be also advantageous to select primers containing C and G nucleotides in a proportion sufficient to ensure efficient hybridization; e.g., an amount of C and G nucleotides of at least 40%, preferably 50% of the total nucleotide amount.
Useful homologs that do not naturally occur can be designed using known methods for 2o identifying regions of an antigen that are likely to be tolerant of amino acid sequence changes and/or deletions. For example, sequences of the antigen from different species can be compared to identify conserved sequences.
Polypeptide derivatives that are encoded by polynucleotides of the invention include, e.g., fragments, polypeptides having large internal deletions derived from full-length polypeptides, and 25 fusion proteins.
Polypeptide fragments of the invention can be derived from a polypeptide having a sequence homologous to any of the sequences shown in SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14 or 16, to the extent that the fragments retain the desired substantial antigenicity of the parent polypeptide (specific antigenicity). Polypeptide derivatives can also be constructed by large internal deletions 3o that remove a substantial part of the parent polypeptide, while retaining the desired specific antigenicity. Generally, polypeptide derivatives should be about at least 12 amino acids in length to maintain the antigenicity. Advantageously, they can be at least 20 amino acids, preferably_at least SO amino acids, more preferably at least 75 amino acids, and most preferably at least 100 amino acids in length.
Useful polypeptide derivatives, e.g., polypeptide fragments, can be designed using computer-assisted analysis of amino acid sequences in order to identify sites in protein antigens having potential as surface-exposed, antigenic regions. Hughes et al., Infect.
Immun. 60: 3497 ( 1992).
Polypeptide fragments and polypeptides having large internal deletions can be used for revealing epitopes that are otherwise masked in the parent polypeptide and that may be of importance for inducing, for example, a protective T cell-dependent immune response. Deletions can also remove inununodorninant regions of high variability among strains.
It is an accepted practice in the field of immunology to use fragments and variants of protein immunogens as vaccines and immunogens, as all that is required to induce an immune response to a protein may be a small (e.g., 8 to 10 amino acid) region of the protein. This has been done for a number of vaccines against pathogens other than Chlamydia. For example, short synthetic peptides corresponding to surface-exposed antigens of pathogens such as murine mammary tumor virus, peptide containing 11 amino acids (Dion et al., Virology 179: 474-477 (1990)); Semliki Forest virus, peptide containing 16 amino acids (Snijders et al., J. Gen. Virol. 72:
557-565 (1991)); and canine parvovirus, two overlapping peptides, each containing 15 amino acids (Langeveld et al., Vaccine 12: 1473-1480 (1994)) have been shown to be effective vaccine antigens against their respective pathogens.
Polynucleotides encoding polypeptide fragments and polypeptides having large internal deletions can be constructed using standard methods (see, e.g., Ausubel et al., CURRENT
PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons Inc. (1994)); for example, by PCR, including inverse PCR, by restriction enzyme treatment of the cloned DNA
molecules, or by the method of Kunkel et al. (Proc. Natl. Acad. Sci. USA 82: 448 ( 1985));
biological material available at Stratagene.
A polypeptide derivative can also be produced as a fusion polypeptide that contains a 3o polypeptide or a polypeptide derivative of the invention fused, e.g., at the N- or C-terminal end, to any other polypeptide. For construction of DNA encoding the amino acid sequence corresponding to hybrid fusion proteins, a first DNA encoding amino acid sequence corresponding to any of the portions of CPN100111, CPN 100224, CPN 100230, CPN 100231, CPN 100232, CPN
100233, CPN 100394, or CPN 100395 nucleotide sequence (SEQ ID NOS: l, 3, 5, 7, 9, 11, 13 or 15) is joined to a second DNA using methods described in, for example, U.S. Patent 5,844,095, incorporated herein by reference. A product can then be easily obtained by translation of the genetic fusion. Vectors for expressing fusion polypeptides are commercially available, such as the pMal-c2 or pMal-p2 systems of New England Biolabs, in which the fusion peptide is a maltose binding protein, the glutathione-S-transferase system of Pharmacia, or the His-Tag system Io available from Novagen. These and other expression systems provide convenient means for further purification of polypeptides and derivatives of the invention.
Another particular example of fusion polypeptides included in the invention includes a polypeptide or polypeptide derivative of the invention fused to a polypeptide having adjuvant activity, such as, e.g., the subunit B of either cholera toxin or E. coli heat-labile toxin. Several possibilities are can be used for achieving fusion. First, the polypeptide of the invention can be fused to the N-, or preferably, to the C-terminal end of the polypeptide having adjuvant activity.
Second, a polypeptide fragment of the invention can be fused within the amino acid sequence of the polypeptide having adjuvant activity.
As stated above, the polynucleotides of the invention encode Chlamydia polypeptides in 2o precursor or mature form. They can also encode hybrid precursors containing heterologous signal peptides, which can mature into polypeptides of the invention. By "heterologous signal peptide" is meant a signal peptide that is not found in the naturally-occurnng precursor of a polypeptide of the invention.
A polynucleotide of the invention, having_a homologous coding sequence, hybridizes, preferably under stringent conditions, to a polynucleotide having a sequence as shown in SEQ ID
NOS: 2, 4, 6, 8, 10, 12, 14 and i6. Hybridization procedures are described in, e.g., Ausubel et al., CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons Inc. (1994); Silhavy et al., EXPERIMENTS WITH GENE FUSIONS, Cold Spring Harbor Laboratory Press (1984);
Davis et al., A
MANUAL FOR GENETIC ENGINEERING: ADVANCED BACTERIAL GENETICS, Cold Spring Harbor 3o Laboratory Press (1980), each incorporated herein by reference. Important parameters that can be considered for optimizing hybridization conditions are reflected in a formula that allows calculation of a critical value, the melting temperature above which two complementary DNA
strands separate from each other. Casey and Davidson, Nucl. Acid Res. 4: 1539 (1977). This formula is as follows:
Tm = 81.5 + 0.5 x (% G+C) + 1.6 log (positive ion concentration) - 0.6 x (%
formamide).
Under appropriate stringency conditions, hybridization temperature (Th) is approximately 20-40°C, 20-25°C or, preferably, 30-40°C below the calculated Tm. Those skilled in the art will understand that optimal temperature and salt conditions can be readily determined empirically in preliminary experiments using conventional procedures.
to For example, stringent conditions can be achieved, both for pre-hybridizing and hybridizing incubations, (i) within 4-16 hours at 42°C, in 6xSSC
containing 50% formamide or (ii) within 4-16 hours at 65°C in an aqueous 6xSSC solution (1 M NaCI, 0.1 M
sodium citrate (pH 7.0)).
For polynucleotides containing 30 to 600 nucleotides, the above formula is used and then is 15 corrected by subtracting (600/polynucleotide size in base pairs).
Stringency conditions are defined by a Th that is 5 to 10°C below Tm.
Hybridization conditions with oligonucleotides shorter than 20-30 bases do not exactly follow the rules set forth above. In such cases, the formula for calculating the Tm is as follows:
Tm = 4 x (G+C) + 2 (A+T).
20 For example, an 18 nucleotide fragment of 50% G+C would have an approximate Tm of 54°C.
A polynucleotide molecule of the invention, containing RNA, DNA, or modifications or combinations thereof, can have various applications. For example, a DNA
molecule can be used:
(i) in a process for producing the encoded polypeptide in a recombinant host system, (ii) in the construction of vaccine vectors such as poxviruses, which are further used in methods and 25 compositions for preventing and/or treating Chlamydia infection, (iii) as a vaccine agent (as well as an RNA molecule), in a naked form or formulated with a delivery vehicle and, (iv) in the construction of attenuated Chlamydia strains that can overexpress a polynucleotide of the invention or express it in a modified, mutated form, such as a non-toxic form, if appropriate.

For vaccine compositions and uses of the proteins and peptides and encoding nucleotides of the present invention for protection against diseases caused by Chlamydia, it is not preferred to use naked DNA encoding the protein or peptides and administering these nucleotides intranasally or intramuscularly. For these proteins, it is preferred to administer the encoding nucleic acids by other routes such as intradermally and/or to formulate the encoding nucleic acids to improve (or adjuvant) the immune response. It is also preferred to include the encoding nucleic acid as part of a recombinant live vector, such as a viral or bacterial vector for use as the immunizing agent. It is also preferred to immunize with vaccine formulations comprising the proteins or peptides of the invention themselves. These vaccine formulations may include the use of adjuvants.
to According to a second aspect of the invention, there is therefore provided:
(i) an expression cassette containing a DNA molecule of the invention placed under the control of the elements required for expression, in particular under the control of an appropriate promoter; (ii) an expression vector containing an expression cassette of the invention; (iii) a prokaryotic or eukaryotic cell transformed or transfected with an expression cassette and/or vector of the 15 invention, as well as (iv) a process for producing a polypeptide or polypeptide derivative encoded by a polynucleotide of the invention, which involves culturing a prokaryotic or eukaryotic cell transformed or transfected with an expression cassette and/or vector of the invention, under conditions that allow expression of the DNA molecule of the invention and, recovering the encoded polypeptide or polypeptide derivative from the cell culture.
2o A recombinant expression system can be selected from prokaryotic and eukaryotic hosts.
Eukaryotic hosts include yeast cells (e.g., Saccharomyces cerevisiae or Pichia pastoris), mammalian cells (e.g., COS1, NIH3T3, or JEG3 cells), arthropods cells (e.g., Spodoptera frugiperda (SF9) cells), and plant cells. Preferably, a prokaryotic host such as E. toll is used.
Bacterial and eukaryotic cells are available from a number of different sources to those skilled in 25 the art, e.g., the American Type Culture Collection (ATCC; Rockville, Maryland).
The choice of the expression system depends on the features desired for the expressed polypeptide. For example, it may be useful to produce a polypeptide of the invention in a particular lipidated form or any other form.
The choice of the expression cassette will depend on the host system selected as well as the 3o features desired for the expressed polypeptide. Typically, an expression cassette includes a promoter that is functional in the selected host system and can be constitutive or inducible; a ribosome binding site; a start codon (ATG) if necessary, a region encoding a signal peptide, e.g., a lipidation signal peptide; a DNA molecule of the invention; a stop codon; and optionally a 3' terminal region (translation and/or transcription terminator}. The signal peptide encoding region is adjacent to the polynucleotide of the invention and placed in proper reading frame. The signal peptide-encoding region can be homologous or heterologous to the DNA molecule encoding the mature polypeptide and can be specific to the secretion apparatus of the host used for expression.
The open reading frame constituted by the DNA molecule of the invention, solely or together with the signal peptide, is placed under the control of the promoter so that transcription and translation occur in the host system. Promoters, signal peptide encoding regions are widely known and available to those skilled in the art and includes, for example, the promoter of Salmonella typhimurium {and derivatives) that is inducible by arabinose (promoter araB) and is functional in Gram-negative bacteria such as E coli (as described in U.S. Patent 5,028,530 and in Cagnon et al., (Cagnon et al., Protein Engineering ~: 843 ( 1991 )); the promoter of the gene of bacteriophage T7 IS encoding RNA polymerase, that is functional in a number of E. coli strains expressing T7 polymerase (described in U.S. Patent 4,952,496); OspA lipidation signal peptide; and RIpB
lipidation signal peptide (Takase et al., J. Bact. 169: 5692 (1987)).
The expression cassette is typically part of an expression vector, which is selected for its ability to replicate in the chosen expression system. Expression vectors (e.g., plasmids or viral vectors) can be chosen from those described in Pouwels et al. (CLONING
VECTORS: LABORATORY
MANUAL, 85, Supp. 1987). They can be purchased from various commercial sources.
Methods for transforming/transfecting host cells with expression vectors will depend on the host system selected as described in Ausubel et al., CURRENT PROTOCOLS IN
MOLECULAR
BIOLOGY, John Wiley & Sons Inc. (1994).
Upon expression, a recombinant polypeptide of the invention (or a polypeptide derivative) is produced and remains in the intracellular compartment, is secreted/excreted in the extracellular medium or in the periplasmic space, or is embedded in the cellular membrane.
The polypeptide can then be recovered in a substantially purified form from the cell extract or from the supernatant after centrifugation of the recombinant cell culture. Typically, the recombinant polypeptide can be 3o purified by antibody-based affinity purification or by any other method that can be readily adapted by a person skilled in the art, such as by genetic fusion to a small affinity binding domain.
Antibody-based affinity purification methods are also available for purifying a polypeptide of the invention extracted from a Chlamydia strain. Antibodies useful for purifying by immunoaffinity the polypeptides of the invention can be obtained as described below.
A polynucleotide of the invention can also be useful in the vaccine field, e.g., for achieving DNA vaccination. There are two major possibilities, either using a viral or bacterial host as gene delivery vehicle (live vaccine vector) or administering the gene in a free form, e.g., inserted into a plasmid. Therapeutic or prophylactic efficacy of a polynucleotide of the invention can be evaluated as described below.
1 o Accordingly, in a third aspect of the invention, there is provided: (i) a vaccine vector such as a poxvirus, containing a DNA molecule of the invention, placed under the control of elements required for expression; (ii) a composition of matter containing a vaccine vector of the invention, together with a diluent or earner; particularly, (iii) a pharmaceutical composition containing a therapeutically or prophylactically effective amount of a vaccine vector of the invention; (iv) a 15 method for inducing an immune response against Chlamydia in a mammal (e.g., a human;
alternatively, the method can be used in veterinary applications for treating or preventing Chlamydia infection of animals, e.g., cats or birds), which involves administering to the mammal an immunogenically effective amount of a vaccine vector of the invention to elicit an immune response, e.g., a protective or therapeutic immune response to Chlamydia; and particularly, (v) a 2o method for preventing and/or treating a Chlamydia (e.g., C. trachomatis, C.
psittaci, C. pneumonia, C. pecorum) infection, which involves administering a prophylactic or therapeutic amount of a vaccine vector of the invention to an individual in need.
Additionally, the third aspect of the invention encompasses the use of a vaccine vector of the invention in the preparation of a medicament for preventing and/or treating Chlamydia infection.
25 A vaccine vector of the invention can express one or several polypeptides or derivatives of the invention, as well as at least one additional Chlamydia antigen, fragment, homolog, mutant, or derivative thereof. In addition, it can express a cytokine, such as interleukin-2 (IL-2) or interleukin-12 (IL-12), that enhances the immune response (adjuvant effect).
Thus, a vaccine vector can include an additional DNA sequence encoding, e.g., a chlamydial antigen , or a 3o cytokine, placed under the control of elements required for expression in a mammalian cell.
-1$-Alternatively, a composition of the invention can include several vaccine vectors, each of them being capable of expressing a polypeptide or derivative of the invention.
A composition can also contain a vaccine vector capable of expressing an additional Chlamydia antigen, or a subunit, fragment, homolog, mutant, or derivative thereof; or a cytokine such as IL-2 or IL-12.
In vaccination methods for treating or preventing infection in a mammal, a vaccine vector of the invention can be administered by any conventional route in use in the vaccine field, particularly, to a mucosal (e.g., ocular, intranasal, oral, gastric, pulmonary, intestinal, rectal, vaginal, or urinary tract) surface or via the parenteral route (e.g., subcutaneous, intradermal, intramuscular, intravenous, or intraperitoneal). Preferred routes depend upon the choice of the vaccine vector. The administration can be achieved in a single dose or repeated at intervals. The appropriate dosage depends on various parameters understood by skilled artisans such as the vaccine vector itself, the route of administration or the condition of the mammal to be vaccinated (weight, age and the like).
Live vaccine vectors available in the art include viral vectors such as adenoviruses, alphavirus, and poxviruses as well as bacterial vectors, e.g., Shigella, Salmonella, Vibrio cholerae, Lactobacillus, Bacille bike de Calmette-Guerin (BCG), and Streptococcus.
An example of an adenovirus vector, as well as a method for constructing an adenovirus vector capable of expressing a DNA molecule of the invention, are described in U.S. Patent 4,920,209. Poxvirus vectors that can be used include, e.g., vaccinia and canary pox virus, 2o described in U.S. Patent 4,722,848 and U.S. Patent 5,364,773, respectively (also see, e.g., Tartaglia et al., Virology 188: 217 (1992)) for a description of a vaccinia virus vector; and Taylor et al, Vaccine 13: S39 (1995) for a reference of a canary pox). Poxvirus vectors capable of expressing a polynucleotide of the invention can be obtained by homologous recombination as described in Kieny et al., Nature 312: 163 ( 1984) so that the polynucleotide of the invention is inserted in the viral genome under appropriate conditions for expression in mammalian cells.
Generally, the dose of vaccine viral vector, for therapeutic or prophylactic use, can be of from about 1x10" to about 1 x 10", advantageously from about 1 x 10' to about 1 x 10' °, preferably of from about 1 x 10' to about 1x109 plaque-forming units per kilogram. Preferably, viral vectors are administered parenterally, for example, in three doses, four weeks apart. Those skilled in the art recognize that it is preferable to avoid adding a chemical adjuvant to a composition containing a viral vector of the invention and thereby minimizing the immune response to the viral vector itself.
Non-toxicogenic Vibrio cholerae mutant strains that are useful as a live oral vaccine are described in Mekalanos et al., Nature 306: 551 (1983) and U.S. Patent 4,882,278 (strain in which a substantial amount of the coding sequence of each of the two ctxA alleles has been deleted so that no functional cholerae toxin is produced); WO 92/11354 (strain in which the irgA locus is inactivated by mutation; this mutation can be combined in a single strain with ctxA mutations); and WO 94/1533 (deletion mutant lacking functional ctxA and attRSl DNA sequences).
These strains can be genetically engineered to express heterologous antigens, as described in WO 94/19482. An to effective vaccine dose of a Vibrio cholerae strain capable of expressing a polypeptide or polypeptide derivative encoded by a DNA molecule of the invention can contain, e.g., about 1x105 to about 1 x 1 O9, preferably about 1 x 1 O6 to about 1 x 108 viable bacteria in an appropriate volume for the selected route of administration. Preferred routes of administration include all mucosal routes, most preferably, these vectors are administered intranasally or orally.
Attenuated Salmonella typhimurium strains, genetically engineered for recombinant expression of heterologous antigens or not, and their use as oral vaccines are described in Nakayama et al., BiolTechnolog~ 6: 693 (1988) and WO 92/11361. Preferred routes of administration include all mucosal routes; most preferably, these vectors are administered intranasally or orally.
2o Others bacterial strains useful as vaccine vectors are described in High et al., EMBO 11:
1991 (1992); Sizemore et al., Science 270: 299 (1995) (ShigellaJlexneri);
Medaglini et al., Proc.
Natl. Acad Sci. USA 92: 6868 (1995) (Streptococcusgordonii); and Flynn, Cell.
Mol. Biol. 40: 31 (1994), WO 88/6626, WO 90/0594, WO 91/13157, WO 92/1796, and WO 92/21376 (Bacille Calmette Guerin).
In bacterial vectors, polynucleotide of the invention can be inserted into the bacterial genome or can remain in a free state, carried on a plasmid.
An adjuvant can also be added to a composition containing a vaccine bacterial vector. A
number of adjuvants are known to those skilled in the art. Preferred adjuvants can be selected from the list provided below.

According to a fourth aspect of the invention, there is also provided: (i) a composition of matter containing a polynucleotide of the invention, together with a diluent or carrier; (ii) a pharmaceutical composition containing a therapeutically or prophylacticaily effective amount of a polynucleotide of the invention; (iii) a method for inducing an immune response against Chlamydia, in a mammal, by administering to the mammal, an imrnunogenically effective amount of a polynucleotide of the invention to elicit an immune response, e.g., a protective immune response to Chlamydia; and particularly, (iv) a method for preventing and/or treating a Chlamydia (e.g., C. trachomatis, C. psittaci, C. pneumoniae, or C. pecorum) infection, by administering a prophylactic or therapeutic amount of a polynucleotide of the invention to an individual in need.
Additionally, the fourth aspect of the invention encompasses the use of a polynucleotide of the invention in the preparation of a medicament for preventing and/or treating Chlamydia infection.
The fourth aspect of the invention preferably includes the use of a DNA
molecule placed under conditions for expression in a mammalian cell, e.g., in a plasmid that is unable to replicate in mammalian cells and to substantially integrate in a mammalian genome.
Polynucleotides (DNA or RNA) of the invention can also be administered as such to a mammal for vaccine, e.g., therapeutic or prophylactic, purpose. When a DNA
molecule of the invention is used, it can be in the form of a plasmid that is unable to replicate in a mammalian cell and unable to integrate in the mammalian genome. Typically, a DNA molecule is placed under the control of a promoter suitable for expression in a mammalian cell. The promoter can function 2o ubiquitously or tissue-specifically. Examples of non-tissue specific promoters include the early Cytomegalovirus (CMV) promoter (described in U.S. Patent 4,168,062) and the Rous Sarcoma Virus promoter (described in Norton & Coffin, Molec. Cell Biol. ~: 281(1985)).
The desmin promoter (Li et al., Gene 78: 243 (1989), Li & Paulin, J. Biol. Chem. 266:
6562 (1991), and Li &
Paulin, J. Biol. Chem. 268: 10403 (1993)) is tissue-specific and drives expression in muscle cells.
More generally, useful vectors are described, i.a., WO 94/21797 and Hartikka et al., Human Gene Therapy 7: 1205 (1996).
For DNA/RNA vaccination, the polynucleotide of the invention can encode a precursor or a mature form. When it encodes a precursor form, the precursor form can be homologous or heterologous. In the latter case, a eukaryotic leader sequence can be used, such as the leader 3o sequence of the tissue-type plasminogen factor (tPA).

A composition of the invention can contain one or several polynucleotides of the invention.
It can also contain at least one additional polynucleotide encoding another Chlamydia antigen or a fragment, derivative, mutant, or analog thereof. A polynucleotide encoding a cytokine, such as interleukin-2 (IL-2) or interleukin-12 (IL-12), can also be added to the composition so that the immune response is enhanced. These additional polynucleotides are placed under appropriate control for expression. Advantageously, DNA molecules of the invention and/or additional DNA
molecules to be included in the same composition, can be carried in the same plasmid.
Standard techniques of molecular biology for preparing and purifying polynucleotides can be used in the preparation of poiynucleotide therapeutics of the invention.
For use as a vaccine, a l0 polynucleotide of the invention can be formulated according to various methods.
First, a polynucleotide can be used in a naked form, free of any delivery vehicles, such as anionic liposomes, cationic lipids, microparticles, e.g., gold microparticles, precipitating agents, e.g., calcium phosphate, or any other transfection-facilitating agent. In this case, the polynucleotide can be simply diluted in a physiologically acceptable solution, such as sterile saline 15 or sterile buffered saline, with or without a carrier. When present, the earner preferably is isotonic, hypotonic, or weakly hypertonic, and has a relatively low ionic strength, such as provided by a sucrose solution, e.g., a solution containing 20% sucrose.
Alternatively, a polynucleotide can be associated with agents that assist in cellular uptake.
It can be, i.a., (i) complemented with a chemical agent that modifies the cellular permeability, such 2o as bupivacaine (see, e.g., WO 94/16737), (ii) encapsulated into liposomes, or (iii) associated with cationic lipids or silica, gold, or tungsten microparticles.
Anionic and neutral liposomes are well-known in the art (see, e.g., LIPOSOMES:
A
PRACTICAL APPROACH, RPC New Ed, IRL press (1990)), for a detailed description of methods for making liposomes) and are useful for delivering a. large range of products, including 25 polynucleotides.
Cationic lipids are also known in the art and are commonly used for gene delivery. Such lipids include LipofectinT'~'' also known as DOTMA (N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride), DOTAP (1,2-bis(oleyloxy)-3-(trimethylammonio)propane), DDAB
(dimethyldioctadecylammonium bromide), DOGS (dioctadecylamidologlycyl spermine) and 3o cholesterol derivatives such as DC-Chol (3 beta-(N-(N',N'-dimethyl aminomethane)-carbamoyl) cholesterol). A description of these cationic lipids can be found in EP
187,702, WO 90/11092, U.S. Patent 5,283,185, WO 91/15501, WO 95/26356, and U.S. Patent 5,527,928.
Cationic lipids for gene delivery are preferably used in association with a neutral lipid such as DOPE (dioleyl phosphatidylethanolamine), as described in, e.g., WO 90/11092.
Other transfection-facilitating compounds can be added to a formulation containing cationic liposomes. A number of them are described in, e.g., WO 93/18759, WO
93/19768, WO 94/25608, and WO 95/2397. They include, i.a., spermine derivatives useful for facilitating the transport of DNA through the nuclear membrane (see, for example, WO
93/18759) and membrane-permeabilizing compounds such as GALA, Gramicidine S, and cationic bile salts (see, to for example, WO 93/19768).
Gold or tungsten microparticles can also be used for gene delivery, as described in WO 91/359, WO 93/17706, and Tang et al. (Nature 356: 152 (1992)). In this case, the microparticle-coated polynucleotides can be injected via intradermal or intra-epidermal routes using a needleless injection device ("gene gun"), such as those described in U.S. Patents 4,945,050 15 and 5,015,580, and WO 94/24263.
The amount of DNA to be used in a vaccine recipient depends, e.g., on the strength of the promoter used in the DNA construct, the immunogenicity of the expressed gene product, the condition of the mammal intended for administration (e.g., the weight, age, and general health of the mammal), the mode of administration, and the type of formulation. In general, a 2o therapeutically or prophylactically effective dose from about 1 ~g to about 1 mg, preferably, from about 10 pg to about 800 ~g and, more preferably, from about 25 ~g to about 250 fig, can be administered to human adults. The administration can be achieved in a single dose or repeated at intervals.
The route of administration can be any conventional route used in the vaccine field. As 25 general guidance, a polynucleotide of the invention can be administered via a mucosal surface, e.g., an ocular, intranasal, pulmonary, oral, intestinal, rectal, vaginal, and urinary tract surface; or via a parenteral route, e.g., by an intravenous, subcutaneous, intraperitoneal, intradermal, intra-epidermal, or intramuscular route. The choice of the administration route will depend on, e.g., the formulation that is selected. A polynucleotide formulated in association with bupivacaine is 3o advantageously administered into muscles. When a neutral or anionic liposome or a cationic lipid, such as DOTMA or DC-Chol, is used, the formulation can be advantageously injected via intravenous, intranasal (aerosolization), intramuscular, intradermal, and subcutaneous routes. . A
polynucleotide in a naked form can advantageously be administered via the intramuscular, intradermal, or subcutaneous routes.
Although not absolutely required, such a composition can also contain an adjuvant. If so, a systemic adjuvant that does not require concomitant administration in order to exhibit an adjuvant effect is preferable such as, e.g., QS21, which is described in U.S. Patent 5,057,546.
The sequence information provided in the present application enables the design of specific nucleotide probes and primers that can be useful in diagnosis. Accordingly, in a fifth aspect of the t0 invention, there is provided a nucleotide probe or primer having a sequence found in or derived by degeneracy of the genetic code from a sequence shown in SEQ ID NOS: 1, 3, 5, 7, 9, 1 l, 13 and 15.
The term "probe" as used in the present application refers to DNA (preferably single stranded) or RNA molecules (or modifications or combinations thereof) that hybridize under the 15 stringent conditions, as defined above, to nucleic acid molecules having sequences homologous to those shown in SEQ ID NOS: 1, 3, 5, 7, 9, 11, 13 and 15, or to a complementary or anti-sense sequence. Generally, probes are significantly shorter than full-length sequences shown in SEQ ID
NOS: 1, 3, 5, 7, 9, 11, 13 and 15; for example, they can contain from about 5 to about 100, preferably from about 10 to about 80 nucleotides. In particular, probes have sequences that are at 20 least 75%, preferably at least 85%, more preferably 95% homologous to a portion of a sequence as shown in SEQ ID NOS: 1, 3, 5, 7, 9, 11, 13 and 15 or that are complementary to such sequences.
Probes can contain modified bases such as inosine, methyl-5-deoxycytidine, deoxyuridine, dimethylamino-5-deoxyuridine, or diamino-2, 6-purine. Sugar or phosphate residues can also be modified or substituted. For example, a deoxyribose residue can be replaced by a polyamide 25 (Nielsen et al., Science 254: 1497 ( 1991 )) and phosphate residues can be replaced by ester groups such as diphosphate, alkyl, arylphosphonate and phosphorothioate esters. In addition, the 2'-hydroxyl group on ribonucleotides can be modified by including, e.g., alkyl groups.
Probes of the invention can be used in diagnostic tests, as capture or detection probes.
Such capture probes can be conventionally immobilized on a solid support, directly or indirectly, 3o by covalent means or by passive adsorption. A detection probe can be labelled by a detection marker selected from radioactive isotopes; enzymes such as peroxidase, alkaline phosphatase, and enzymes able to hydrolyze a chromogenic, fluorogenic, or luminescent substrate; compounds.that are chromogenic, fluorogenic, or luminescent; nucleotide base analogs; and biotin.
Probes of the invention can be used in any conventional hybridization technique, such as dot blot (Maniatis et al., MOLECULAR CLONING: A LABORATORY MANUAL (1982) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York), Southern blot (Southern, J. Mol. Biol.
98: 503 (1975)), northern blot (identical to Southern blot to the exception that RNA is used as a target), or the sandwich technique (Dune et al., Cell 12: 23 ( 1977)). The latter technique involves the use of a specific capture probe and/or a specific detection probe with nucleotide sequences that I o at least partially differ from each other.
A primer is usually a probe of about 10 to about 40 nucleotides that is used to initiate enzymatic polymerization of DNA in an amplification process (e.g., PCR), in an elongation process, or in a reverse transcription method. In a diagnostic method involving PCR, primers can be labelled.
I S Thus, the invention also encompasses: (i) a reagent containing a probe of the invention for detecting and/or identifying the presence of Chlamydia in a biological material; (ii) a method for detecting and/or identifying the presence of Chlamydia in a biological material, in which (a) a sample is recovered or derived from the biological material, (b) DNA or RNA is extracted from the material and denatured, and (c) exposed to a probe of the invention, for example, a capture, 2o detection probe or both, under stringent hybridization conditions, such that hybridization is detected; and (iii) a method for detecting and/or identifying the presence of Chlamydia in a biological material, in which (a) a sample is recovered or derived from the biological material, (b) DNA is extracted therefrom, (c) the extracted DNA is primed with at least one, and preferably two, primers of the invention and amplified by polymerase chain reaction, and (d) the amplified 25 DNA fragment is produced.
As previously mentioned, polypeptides that can be produced upon expression of the newly identified open reading frames are useful vaccine agents.
Therefore, a sixth aspect of the invention features a substantially purified polypeptide or polypeptide derivative having an amino acid sequence encoded by a polynucleotide of the 30 invention.

A "substantially purified polypeptide" is defined as a polypeptide that is separated from the environment in which it naturally occurs and/or that is free of the majority of the polypeptides that are present in the environment in which it was synthesized. For example, a substantially purified polypeptide is free from cytoplasmic polypeptides. Those skilled in the art will understand that the polypeptides of the invention can be purified from a natural source, i.e., a Chlamydia strain, or can be produced by recombinant means.
Homologous polypeptides or polypeptide derivatives encoded by polynucleotides of the invention can be screened for specific antigenicity by testing cross-reactivity with an antiserum raised against the polypeptide of reference having an amino acid sequence as shown in SEQ ID
NOS: 2, 4, 6, 8, 10, 12, 14 and 16. Briefly, a monospecific hyperimmune antiserum can be raised against a purified reference polypeptide as such or as a fusion polypeptide, for example, an expression product of MBP, GST, or His-tag systems or a synthetic peptide predicted to be antigenic. The homologous polypeptide or derivative screened for specific antigenicity can be produced as such or as a fusion polypeptide. In this latter case and if the antiserum is also raised against a fusion polypeptide, two different fusion systems are employed.
Specific antigenicity can be determined according to a number of methods, including Western blot (Towbin et al., Proc.
Natl. Acad. SCI. USA 76: 4350 (1979)), dot blot, and ELISA, as described below.
In a Western blot assay, the product to be screened, either as a purified preparation or a total E. toll extract, is submitted to SDS-Page electrophoresis as described by Laemmli, Nature 227: 680 (1970). After transfer to a nitrocellulose membrane, the material is further incubated with the monospecific hyperimmune antiserum diluted in the range of dilutions from about 1:5 to about 1:5000, preferably from about 1:100 to about 1:500. Specific antigenicity is shown once a band corresponding to the product exhibits reactivity at any of the dilutions in the above range.
In an ELISA assay, the product to be screened is preferably used as the coating antigen. A
purified preparation is preferred, although a whole cell extract can also be used. Briefly, about 100 pl of a preparation at about 10 pg protein/ml are distributed into wells of a 96-well polycarbonate ELISA plate. The plate is incubated for 2 hours at 37°C
then overnight at 4°C. The plate is washed with phosphate buffer saline (PBS) containing 0.05% Tween 20 (PBS/Tween buffer). The wells are saturated with 250 ~.l PBS containing 1% bovine serum albumin (BSA) to 3o prevent non-specific antibody binding. After 1 hour incubation at 37°C, the plate is washed with PBS/Tween buffer. The antiserum is serially diluted in PBS/Tween buffer containing 0.5% BSA.
100 ~1 of dilutions are added per well. The plate is incubated for 90 minutes at 37°C, washed_and evaluated according to standard procedures. For example, a goat anti-rabbit peroxidase conjugate is added to the wells when specific antibodies were raised in rabbits.
Incubation is carried out for 90 minutes at 37°C and the plate is washed. The reaction is developed with the appropriate substrate and the reaction is measured by colorimetry (absorbance measured spectrophotometrically). Under the above experimental conditions, a positive reaction is shown by O.D. values greater than a non immune control serum.
In a dot blot assay, a purified product is preferred, although a whole cell extract can also be to used. Briefly, a solution of the product at about 100 ~g/ml is serially two-fold diluted in SO mM
Tris-HC1 (pH 7.5). 100 ~1 of each dilution are applied to a nitrocellulose membrane 0.45 ~.m set in a 96-well dot blot apparatus (Biorad). The buffer is removed by applying vacuum to the system.
Wells are washed by addition of 50 mM Tris-HCl (pH 7.5) and the membrane is air-dried. The membrane is saturated in blocking buffer (50 mM Tris-HCl (pH 7.5) 0. i 5 M
NaCI, 10 g/L skim milk) and incubated with an antiserum dilution from about I :50 to about 1:5000, preferably about 1:500. The reaction is revealed according to standard procedures. For example, a goat anti-rabbit peroxidase conjugate is added to the wells when rabbit antibodies are used.
Incubation is earned out 90 minutes at 37°C and the blot is washed. The reaction is developed with the appropriate substrate and stopped. The reaction is measured visually by the appearance of a colored spot, e.g., 2o by colorimetry. Under the above experimental conditions, a positive reaction is shown once a colored spot is associated with a dilution of at least about 1:5, preferably of at least about I :500.
Therapeutic or prophylactic efficacy of a polypeptide or derivative of the invention can be evaluated as described below.
According to a seventh aspect of the invention, there is provided: (i) a composition of matter containing a polypeptide of the invention together with a diluent or earner; in particular, (ii) a pharmaceutical composition containing a therapeutically or prophylactically effective amount of a polypeptide of the invention; (iii) a method for inducing an immune response against Chlamydia in a mammal, by administering to the mammal an immunogenically effective amount of a polypeptide of the invention to elicit an immune response, e.g., a protective immune response 3o to Chlamydia; and particularly, (iv) a method for preventing and/or treating a Chlamydia (e.g., C. trachomatis. C. psittaci, C. pneumoniae. or C. pecorum) infection, by administering a prophylactic or therapeutic amount of a polypeptide of the invention to an individual in need..
Additionally, the seventh aspect of the invention encompasses the use of a polypeptide of the invention in the preparation of a medicament for preventing and/or treating Chlamydia infection.
The immunogenic compositions of the invention can be administered by any conventional route in use in the vaccine field, in particular to a mucosal (e.g., ocular, intranasal, pulmonary, oral, gastric, intestinal, rectal, vaginal, or urinary tract) surface or via the parenteral (e.g., subcutaneous, intradermal, intramuscular, intravenous, or intraperitoneal) route. The choice of the administration route depends upon a number of parameters, such as the adjuvant associated with the polypeptide.
1o For example, if a mucosal adjuvant is used, the intranasal or oral route will be preferred and if a lipid formulation or an aluminum compound is used, the parenteral route will be preferred. In the latter case, the subcutaneous or intramuscular route is most preferred. The choice can also depend upon the nature of the vaccine agent. For example, a polypeptide of the invention fused to CTB or LTB will be best administered to a mucosal surface.
A composition of the invention can contain one or several polypeptides or derivatives of the invention. It can also contain at least one additional Chlamydia antigen, or a subunit, fragment, homolog, mutant, or derivative thereof.
For use in a composition of the invention, a polypeptide or derivative thereof can be formulated into or with liposomes, preferably neutral or anionic liposomes, microspheres, 2o ISCOMS, or virus-like-particles (VLPs) to facilitate delivery and/or enhance the immune response.
These compounds are readily available to one skilled in the art; for example, see LIPOSOMES: A
PRACTICAL APPROACH (supra).
Adjuvants other than liposomes and the like can also be used and are known in the art. An appropriate selection can conventionally be made .by those skilled in the art, for example, from the list provided below.
Administration can be achieved in a single dose or repeated as necessary at intervals as can be determined by one skilled in the art. For example, a priming dose can be followed by three booster doses at weekly or monthly intervals. An appropriate dose depends on various parameters including the recipient (e.g., adult or infant), the particular vaccine antigen, the route and 3o frequency of administration, the presence/absence or type of adjuvant, and the desired effect (e.g., protection and/or treatment), as can be determined by one skilled in the art.
In general, a vaccine antigen of the invention can be administered by a mucosal route in an amount from about 10 ~g to about 500 mg, preferably from about 1 mg to about 200 mg. For the parenteral route of administration, the dose usually should not exceed about 1 mg, preferably about 100 pg.
s When used as vaccine agents, polynucleotides and polypeptides of the invention can be used sequentially as part of a multistep immunization process. For example, a mammal can be initially primed with a vaccine vector of the invention such as a pox virus, e.g., via the parenteral route, and then boosted twice with the polypeptide encoded by the vaccine vector, e.g., via the mucosal route. In another example, liposomes associated with a polypeptide or derivative of the to invention can also be used for priming, with boosting being carned out mucosally using a soluble polypeptide or derivative of the invention in combination with a mucosal adjuvant (e.g., LT).
A polypeptide derivative of the invention is also useful as a diagnostic reagent for detecting the presence of anti-Chlamydia antibodies, e.g., in a blood sample. Such polypeptides are about 5 to about 80, preferably about 10 to about 50 amino acids in length and can be labeled or unlabeled, I5 depending upon the diagnostic method. Diagnostic methods involving such a reagent are described below.
Upon expression of a DNA molecule of the invention, a polypeptide or polypeptide derivative is produced and can be purified using known laboratory techniques.
For example, the polypeptide or polypeptide derivative can be produced as a fusion protein containing a fused tail 2o that facilitates purification. The fusion product can be used to immunize a small mammal, e.g., a mouse or a rabbit, in order to raise antibodies against the polypeptide or polypeptide derivative (monospecific antibodies). The eighth aspect of the invention thus provides a monospecific antibody that binds to a polypeptide or polypeptide derivative of the invention.
By "monospecific antibody" is meant an antibody that is capable of reacting with a unique 25 naturally-occurring Chlamydia polypeptide. An antibody of the invention can be polyclonal or monoclonal. Monospecific antibodies can be recombinant, e.g., chimeric (e.g., constituted by a variable region of marine origin associated with a human constant region), humanized (a-human immunoglobulin constant backbone together with hypervariable region of animal, e.g., marine, origin), and/or single chain. Both polyclonal and monospecific antibodies can also be in the form 30 of immunoglobulin fragments, e.g., F(ab)'2 or Fab fragments. The antibodies of the invention can be of any isotype, e.g., IgG or IgA, and polyclonal antibodies can be of a single isotype or can contain a mixture of isotypes.
The antibodies of the invention, which are raised to a polypeptide or polypeptide derivative of the invention, can be produced and identified using standard immunological assays, e.g., Western blot analysis, dot blot assay, or ELISA (see, e.g., Coligan et al., CultttirNT PROTOCOLS 1N
IMMUNOLOGY (1994) John Wiley & Sons, Inc., New York, NY). The antibodies can be used in diagnostic methods to detect the presence of a Chlamydia antigen in a sample, such as a biological sample. The antibodies can also be used in affinity chromatography methods for purifying a polypeptide or polypeptide derivative of the invention. As is discussed further below, such t 0 antibodies can be used in prophylactic and therapeutic passive immunization methods.
Accordingly, a ninth aspect of the invention provides: (i) a reagent for detecting the presence of Chlamydia in a biological sample that contains an antibody, polypeptide, or polypeptide derivative of the invention; and {ii) a diagnostic method for detecting the presence of Chlamydia in a biological sample, by contacting the biological sample with an antibody, a polypeptide, or a polypeptide derivative of the invention, such that an immune complex is formed, and by detecting such complex to indicate the presence of Chlamydia in the sample or the organism from which the sample is derived.
Those skilled in the art will understand that the immune complex is formed between a component of the sample and the antibody, polypeptide, or polypeptide derivative, whichever is 2o used, and that any unbound material can be removed prior to detecting the complex. As can be easily understood, a polypeptide reagent is useful for detecting the presence of anti-Chlamydia antibodies in a sample, e.g., a blood sample, while an antibody of the invention can be used for screening a sample, such as a gastric extract or biopsy, for the presence of Chlamydia polypeptides.
For use in diagnostic applications, the reagent (i.e., the antibody, polypeptide, or polypeptide derivative of the invention) can be in a free state or immobilized on a solid support, such as a tube, a bead, or any other conventional support used in the field.
Immobilization can be achieved using direct or indirect means. Direct means include passive adsorption (non-covalent binding) or covalent binding between the support and the reagent. By "indirect means" is meant 3o that an anti-reagent compound that interacts with a reagent is first attached to the solid support.

For example, if a polypeptide reagent is used, an antibody that binds to it can serve as an anti-reagent, provided that it binds to an epitope that is not involved in the recognition of antibodies in biological samples. Indirect means can also employ a ligand-receptor system, for example, a molecule such as a vitamin can be grafted onto the polypeptide reagent and the corresponding receptor can be immobilized on the solid phase. This is illustrated by the biotin-streptavidin system. Alternatively, indirect means can be used, e.g., by adding to the reagent a peptide tail, chemically or by genetic engineering, and immobilizing the grafted or fused product by passive adsorption or covalent linkage of the peptide tail.
According to a tenth aspect of the invention, there is provided a process for purifying, from to a biological sample, a polypeptide or polypeptide derivative of the invention, which involves carrying out antibody-based affinity chromatography with the biological sample, wherein the antibody is a monospecific antibody of the invention.
For use in a purification process of the invention, the antibody can be polyclonal or monospecific, and preferably is of the IgG type. Purified IgGs can be prepared from an antiserum t5 using standard methods (see, e.g., Coligan et al., supra). Conventional chromatography supports, as well as standard methods for grafting antibodies, are disclosed in, e.g., ANTIBODIES: A
LABORATORY MANUAL, D. Lane, E. Harlow, Eds. ( 1988).
Briefly, a biological sample, such as an C. pneumoniae extract, preferably in a buffer solution, is applied to a chromatography material, preferably equilibrated with the buffer used to 2o dilute the biological sample so that the polypeptide or polypeptide derivative of the invention (i.e., the antigen) is allowed to adsorb onto the material. The chromatography material, such as a gel or a resin coupled to an antibody of the invention, can be in batch form or in a column. The unbound components are washed off and the antigen is then eluted with an appropriate elution buffer, such as a glycine buffer or a buffer containing a chaotropic agent, e.g., guanidine HCI, or high salt 25 concentration (e.g., 3 M MgCl2). Eluted fractions are recovered and the presence of the antigen is detected, e.g., by measuring the absorbance at 280 nm.
An antibody of the invention can be screened for therapeutic efficacy as described as follows. According to an eleventh aspect of the invention, there is provided:
(i) a composition of matter containing a monospecific antibody of the invention, together with a diluent or carrier; (ii) a 3o pharmaceutical composition containing a therapeutically or prophylactically effective amount of a monospecific antibody of the invention, and (iii) a method for treating or preventing a Chlamydia (e.g., C. trachomatis, C. psittaci, C. pneumoniae or C. pecorum) infection, by administering a therapeutic or prophylactic amount of a monospecific antibody of the invention to an individual in need. Additionally, the eleventh aspect of the invention encompasses the use of a monospecific antibody of the invention in the preparation of a medicament for treating or preventing Chlamydia infection.
To this end, the monospecific antibody can be polyclonal or monoclonal, preferably of the IgA isotype (predominantly). In passive immunization, the antibody can be administered to a mucosal surface of a mammal, e.g., the gastric mucosa, e.g., orally or intragastrically, advantageously, in the presence of a bicarbonate buffer. Alternatively, systemic administration, not requiring a bicarbonate buffer, can be carried out. A monospecific antibody of the invention can be administered as a single active component or as a mixture with at least one monospecific antibody specific for a different Chlamydia polypeptide. The amount of antibody and the particular regimen used can be readily determined by one skilled in the art.
For example, daily 15 administration of about 100 to 1,000 mg of antibodies over one week, or three doses per day of about 100 to 1,000 mg of antibodies over two or three days, can be an effective regimens for most purposes.
Therapeutic or prophylactic efficacy can be evaluated using standard methods in the art, e.g., by measuring induction of a mucosal immune response or induction of protective and/or 2o therapeutic immunity, using, e.g., the C. pneumoniae mouse model. Those skilled in the art will recognize that the C. pneumoniae strain of the model can be replaced with another Chlamydia strain. For example, the efficacy of DNA molecules and polypeptides from C.
pneumoniae is preferably evaluated in a mouse model using an C. pneumoniae strain.
Protection can be determined by comparing the degree of Chlamydia infection to that of a control group. Protection 25 is shown when infection is reduced by comparison to the control group. Such an evaluation can be made for polynucleotides, vaccine vectors, polypeptides and derivatives thereof, as well as antibodies of the invention.
Adjuvants useful in any of the vaccine compositions described above are as follows.
Adjuvants for parenteral administration include aluminum compounds, such as aluminum 3o hydroxide, aluminum phosphate, and aluminum hydroxy phosphate. The antigen can be precipitated with, or adsorbed onto, the aluminum compound according to standard protocols.
Other adjuvants, such as RIBI (ImmunoChem, Hamilton, MT), can be used in parenteral administration.
Adjuvants for mucosal administration include bacterial toxins, e.g., the cholera toxin (CT), the E. coli heat-labile toxin (LT), the Clostridium di~cile toxin A and the pertussis toxin (PT), or combinations, subunits, toxoids, or mutants thereof. For example, a purified preparation of native cholera toxin subunit B (CTB) can be of use. Fragments, homologs, derivatives, and fusions to any of these toxins are also suitable, provided that they retain adjuvant activity. Preferably, a mutant having reduced toxicity is used. Suitable mutants are described, e.g., in WO 95/17211 (Arg-7-Lys CT mutant), WO 96/6627 (Arg-192-Gly LT mutant), and WO 95/34323 (Arg-9-Lys and Glu-129-Gly PT mutant). Additional LT mutants that can be used in the methods and compositions of the invention include, e.g., Ser-63-Lys, Ala-69-GIy, Glu-110-Asp, and Glu-112-Asp mutants. Other adjuvants, such as a bacterial monophosphoryl lipid A (MPLA) of, e.g., E. coli, Salmonella minnesota, Salmonella typhimurium, or Shigella flexneri; saponins, or I5 polylactide glycolide (PLGA) microspheres, can also be used in mucosal administration.
Adjuvants useful for both mucosal and parenteral administrations include polyphosphazene (WO 95/2415), DC-chol (3 b-(N-(N',N'-dimethyl aminomethane)-carbamoyl) cholesterol (U.S.
Patent 5,283,185 and WO 96/14831) and QS-21 (WO 88/9336).
Any pharmaceutical composition of the invention, containing a polynucleotide, a polypeptide, a polypeptide derivative, or an antibody of the invention, can be manufactured in a conventional manner. In particular, it can be formulated with a pharmaceutically acceptable diluent or carrier, e.g., water or a saline solution such as phosphate buffer saline. In general, a diluent or carrier can be selected on the basis of the mode and route of administration, and standard pharmaceutical practice. Suitable pharmaceutical Garners or diluents, as well as pharmaceutical necessities for their use in pharmaceutical formulations, are described in Remington's Pharmaceutical Sciences, a standard reference text in this field and in the USP/NF.
The invention also includes methods in which Chlamydia infection, are treated by~ oral administration of a Chlamydia polypeptide of the invention and a mucosal adjuvant, in combination with an antibiotic, an antacid, sucralfate, or a combination thereof. Examples of such compounds that can be administered with the vaccine antigen and the adjuvant are antibiotics, including, e.g., macrolides, tetracyclines, and derivatives thereof (specific examples of antibiotics that can be used include azithromycin or doxicyclin or immunomodulators such as cytokines-or steroids. In addition, compounds containing more than one of the above-listed components coupled together, can be used. The invention also includes compositions for carrying out these methods, i.e., compositions containing a Chlamydia antigen (or antigens) of the invention, an adjuvant, and one or more of the above-listed compounds, in a pharmaceutically acceptable carrier or diluent.
Amounts of the above-listed compounds used in the methods and compositions of the invention can readily be determined by one skilled in the art. In addition, one skilled in the art can 1o readily design treatment/immunization schedules. For example, the non-vaccine components can be administered on days 1-14, and the vaccine antigen + adjuvant can be administered on days 7, 14, 21, and 28.
The above disclosure generally describes the present invention. A more complete understanding can be obtained by reference to the following specific examples.
These examples 15 are described solely for purposes of illustration and are not intended to limit the scope of the invention. Changes in form and substitution of equivalents are contemplated as circumstances may suggest or render expedient. Although specific terms have been employed herein, such terms are intended in a descriptive sense and not for purposes of limitation.
EQUIVALENTS
20 From the foregoing detailed description of the specific embodiments of the invention, it should be apparent that unique Chlamydia antigens have been described.
Although particular embodiments have been disclosed herein in detail, this has been done by way of example for purposes of illustration only, and is not intended to be limiting with respect to the scope of the appended claims which follow. In particular, it is contemplated by the inventor that various 25 substitutions, alterations, and modifications may be made to the invention without departing from the spirit and scope of the invention as defined by the claims.

SEQUENCE LISTING
<110> Murdin, Andrew <120> CFiLAMYDIA ANTIGENS AND CORRESPONDING DNA FRAGMENTS
THEREOF
<130> Chlamydia antigens <140>
<141>
<160> 16 <170> Patentln Ver. 2.0 <210> 1 <211> 2550 <212> DNA
<213> Chlamydia sp.
<220>
<221> CDS
<222> (101)..(2461) <400> 1 taggtcttgt taaaaaactt ctttgtccct atttttgtgc ttacctcatt atctgataaa 60 caagctttct atctagggaa gactcttggg aatgctcatc atg cga aat aaa gtt 115 Met Arg Asn Lys Val atc ttg caa ata tct att cta gcg tta atc caa acc cct tta act tta 163 Ile Leu Gln Ile Ser Ile Leu Ala Leu Ile Gln Thr Pro Leu Thr Leu ttt tct act gaa aaa gtt aaa gaa ggc cat gtg gtg gta gac tct atc 211 Phe Ser Thr Glu Lys Val Lys Glu Gly His Val Val Val Asp Ser Ile aca atc ata acg gaa gga gaa aat get tca aat aaa cat ccc tta ccc 259 Thr Ile Ile Thr Glu Gly Glu Asn Ala Ser Asn Lys His Pro Leu Pro aaa tta aag acc aga agt ggg get ctt ttt tct caa tta gat ttt gat 307 Lys Leu Lys Thr Arg Ser Gly Ala Leu Phe Ser Gln Leu Asp Phe Asp gaa ttg attcta aaa tac gactctgtt gagcct aaa 355 gac aga get gaa Glu Leu IleLeu Lys Tyr AspSerVal GluPro Lys Asp Arg Ala Glu gta ttt gaaggg act ata gcccttcac ctaata get 403 gaa tct aaa aac Val Phe GluGly Thr Ile AlaLeuHis LeuIle Ala Glu Ser Lys Asn aaa ccc tca att cga aat att cat atc tca gga aat caa gtc gtt cct 451 Lys Pro Ser Ile Arg Asn Ile His Ile Ser Gly Asn Gln Val Val Pro gaa cat aaa att ctt aaa acc cta caa att tac cgt aat gat ctc ttt 499 Glu His Lys Ile Leu Lys Thr Leu Gln Ile Tyr Arg Asn Asp Leu Phe gaa cga gaa aaa ttt ctt aag ggt ctt gat gat cta aga acg tat tat 547 Glu Arg Glu Lys Phe Leu Lys Gly Leu Asp Asp Leu Arg Thr Tyr Tyr ctc aag cga gga tat ttc gca tcc agt gta gac tac agt ctg gaa cac 595 Leu Lys Arg Gly Tyr Phe Ala Ser Ser Val Asp Tyr Sex Leu Glu His aat caa gaa aaa ggt cac atc gat gtt tta att aaa atc aat gaa ggt 643 Asn Gln Glu Lys Gly His Ile Asp Val Leu Ile Lys Ile Asn Glu Gly cct tgc ggg aaa att aaa cag ctt acg ttc tca gga atc tct cga tca 691 Pro Cys Gly Lys Ile Lys Gln Leu Thr Phe Ser Gly Ile Ser Arg Ser gaa aaa tca gat atc caa gaa ttt att caa acc aag cag cac tct aca 739 Glu Lys Ser Asp Ile Gln Glu Phe Ile Gln Thr Lys Gln His Ser Thr act aca agt tgg ttt act gga get gga ctc tat cac cca gat att gtt 787 Thr Thr Ser Trp Phe Thr Gly Ala Gly Leu Tyr His Pro Asp Ile Val gaa caa gat agc ttg gca att acg aat tac cta cat aat aac ggg tac 835 Glu Gln Asp Ser Leu Ala Ile Thr Asn Tyr Leu His Asn Asn Gly Tyr get gat get ata gtc aac tct cac tat gac ctt gac gac aaa ggg aat 883 Ala Asp Ala Ile Val Asn Ser His Tyr Asp Leu Asp Asp Lys Gly Asn att ctt ctt tac atg gat att gat cga ggg tcg cga tat acc tta gga 931 Ile Leu Leu Tyr Met Asp Ile Asp Arg Gly Ser Arg Tyr Thr Leu Gly cac gtc cat atc caa ggg ttt gag gtt ttg cca aaa cgc ctt ata gaa 979 His Val His Ile Gln Gly Phe Glu Val Leu Pro Lys Arg Leu Ile Glu aag caa tcc caa gtc ggc ccc aat gat ctt tat tgc ccc gat aaa ata 1027 Lys Gln Ser Gln Val Gly Pro Asn Asp Leu Tyr Cys Pro Asp Lys Ile 2g5 300 305 tgg gat ggg get cat aag atc aaa caa act tat gca aag tat ggc tac 1075 Trp Asp Gly Ala His Lys Ile Lys Gln Thr Tyr Ala Lys Tyr Gly Tyr atc aat acc aat gta gac gtt ctc ttc atc cct cac gca acc cgc cct 1123 Ile Asn Thr Asn Val Asp Val Leu Phe Ile Pro His Ala Thr Arg Pro att tat gat gta act tat gag gta agt gaa ggg tct cct tat aaa gtt 1171 Ile Tyr Asp Val Thr Tyr Glu Val Ser Glu Gly Ser Pro Tyr Lys Val ggg tta att aaa att act ggg aat acc cat aca aaa tct gac gtt att 1219 Gly Leu Ile Lys Ile Thr Gly Asn Thr His Thr Lys Ser Asp Val Ile tta cac gaa acc agt ctc ttc cca gga gat aca ttc aat cgc tta aag 1267 Leu His Glu Thr Ser Leu Phe Pro Gly Asp Thr Phe Asn Arg Leu Lys cta gaa gat act gag caa cgt tta aga aat aca ggc tac ttc caa agc 1315 Leu Glu Asp Thr Glu Gln Arg Leu Arg Asn Thr Gly Tyr Phe Gln Ser gtt agt gtc tat aca gtt cgt tct caa ctt gat cct atg ggc aat gcg 1363 Val Ser Val Tyr Thr Val Arg Ser Gln Leu Asp Pro Met Gly Asn Ala gat caa tac cga gat att ttt gta gaa gtc aaa gaa aca aca aca gga 1411 Asp Gln Tyr Arg Asp Ile Phe Val Glu Val Lys Glu Thr Thr Thr Gly aac tta ggc tta ttc tta gga ttt agt tct ctt gac aat ctt ttt gga 1459 Asn Leu Gly Leu Phe Leu Gly Phe Ser Ser Leu Asp Asn Leu Phe Gly gga att gaa cta tct gaa agt aat ttt gat cta ttt gga get aga aat 1507 Gly Ile Glu Leu Ser Glu Ser Asn Phe Asp Leu Phe Gly Ala Arg Asn ata ttt tct aaa ggt ttt cgt tgt cta aga ggc ggt gga gaa cat cta 1555 Ile Phe Ser Lys Gly Phe Arg Cys Leu Arg Gly Gly Gly Glu His Leu ttc tta aaa gcc aac ttc ggg gac aaa gtc aca gac tat act ttg aag 1603 Phe Leu Lys Ala Asn Phe Gly Asp Lys Val Thr Asp Tyr Thr Leu Lys tgg acc aaa cct cat ttt cta aac act cct tgg att tta gga att gaa 1651 Trp Thr Lys Pro His Phe Leu Asn Thr Pro Trp Ile Leu Gly Ile Glu tta gat aaa tca att aac aga gca tta tct aaa gat tat get gtc caa 1699 Leu Asp Lys Ser Ile Asn Arg Ala Leu Ser Lys Asp Tyr Ala Val Gln acc tat ggc ggg aac gtc agc aca acg tat atc ttg aac gaa cac ctg 1747 Thr Tyr Gly Gly Asn Val Ser Thr Thr Tyr Ile Leu Asn Glu His Leu aaa tac ggt cta ttt tat cga gga agt caa acg agt tta cat gaa aaa 1795 Lys Tyr Gly Leu Phe Tyr Arg Gly Ser Gln Thr Ser Leu His Glu Lys cgt aag ttc ctc cta ggg cca aat ata gac agc aat aaa gga ttt gtc 1843 Arg Lys Phe Leu Leu Gly Pro Asn Ile Asp Ser Asn Lys Gly Phe Val tct get gca ggt gtc aac ttg aat tac gat tct gta gat agt cct aga 1891 Ser Ala Ala Gly Val Asn Leu Asn Tyr Asp Ser Val Asp Ser Pro Arg act cca act aca ggg att cgc ggg ggg gtg act ttt gag gtt tct ggt 1939 Thr Pro Thr Thr Gly Ile Arg Gly Gly Val Thr Phe Glu Val Ser Gly ttg gga gga act tat cat ttt aca aaa ctc tct tta aac agc tct atc 1987 Leu Gly Gly Thr Tyr His Phe Thr Lys Leu Ser Leu Asn Ser Ser Ile tat aga aaa ctt acg cgt aaa ggt att ttg aaa atc aaa ggg gaa get 2035 Tyr Arg Lys Leu Thr Arg Lys Gly Ile Leu Lys Ile Lys Gly Glu Ala caa ttt att aaa ece tat agc aat act aca get gaa gga gat ect gtc 2083 Gln Phe Ile Lys Pro Tyr Ser Asn Thr Thr Ala Glu Gly Asp Pro Val agt gag cgc ttc ttc cta ggt gga gag act aca gtt cgg gga tat aaa 2131 Ser Glu Arg Phe Phe Leu Gly Gly Glu Thr Thr Val Arg Gly Tyr Lys tcc ttt att atc ggt cca aaa tac tct get aca gaa cct cag gga gga 2179 Ser Phe Ile Ile Gly Pro Lys Tyr Ser Ala Thr Glu Pro Gln Gly Gly ctc tct tcg ctc ctt att tca gaa gag ttt caa tac cct ctc atc aga 2227 Leu Ser Ser Leu Leu Ile Ser Glu Glu Phe Gln Tyr Pro Leu Ile Arg caa cct aat att agt gcc ttt gta ttc tta gac tca ggt ttt gtc ggt 2275 Gln Pro Asn Ile Ser Ala Phe Val Phe Leu Asp Ser Gly Phe Val Gly tta caa gag tat aag att tcg tta aaa gat cta cgt agt agt get gga 2323 Leu Gln Glu Tyr Lys Ile Ser Leu Lys Asp Leu Arg Ser Ser Ala Gly ttt ggt ctg cgc ttc gat gta atg aat aat gtt cct gtt atg tta gga 2371 Phe Gly Leu Arg Phe Asp Val Met Asn Asn Val Pro Val Met Leu Gly ttt ggt tgg ccc ttc cgt cca acc gag act ttg aat gga gaa aaa att 2419 Phe Gly Trp Pro Phe Arg Pro Thr Glu Thr Leu Asn Gly Glu Lys Ile gat gta tct cag cga ttc ttc ttt get tta ggg ggc atg ttc 2461 Asp Val Ser Gln Arg Phe Phe Phe Ala Leu Gly Gly Met Phe taagatataa attaaggact tatcgaagga aatctttgtt gttttcagaa aaggcttttg 2521 gtaccctttt tcctataccc aaagttagt 2550 <210> 2 <211> 787 <212> PRT
<213> Chlamydia sp.
<400> 2 Met Arg Asn Lys Val Ile Leu Gln Ile Ser Ile Leu Ala Leu Ile Gln Thr Pro Leu Thr Leu Phe Ser Thr Glu Lys Val Lys Glu Gly His Val Val Val Asp Ser Ile Thr Ile Ile Thr Glu Gly Glu Asn Ala Ser Asn Lys His Pro Leu Pro Lys Leu Lys Thr Arg Ser Gly Ala Leu Phe Ser Gln Leu Asp Phe Asp Glu Asp Leu Arg Ile Leu Ala Lys Glu Tyr Asp Ser Val Glu Pro Lys Val Glu Phe Ser Glu Gly Lys Thr Asn Ile Ala Leu His Leu Ile Ala Lys Pro Ser Ile Arg Asn Ile His Ile Ser Gly Asn Gln Val Val Pro Glu His Lys Ile Leu Lys Thr Leu Gln Ile Tyr Arg Asn Asp Leu Phe Glu Arg Glu Lys Phe Leu Lys Gly Leu Asp Asp Leu Arg Thr Tyr Tyr Leu Lys Arg Gly Tyr Phe Ala Ser Ser Val Asp Tyr Ser Leu Glu His Asn Gln Glu Lys Gly His Ile Asp Val Leu Ile Lys Ile Asn Glu Gly Pro Cys Gly Lys Ile Lys Gln Leu Thr Phe Ser Gly Ile Ser Arg Ser Glu Lys Ser Asp Ile Gln Glu Phe Ile Gln Thr Lys Gln His Ser Thr Thr Thr Ser Trp Phe Thr Gly Ala Gly Leu Tyr His Pro Asp Ile Val Glu Gln Asp Ser Leu Ala Ile Thr Asn Tyr Leu His Asn Asn Gly Tyr Ala Asp Ala Ile Val Asn Ser His Tyr Asp Leu Asp Asp Lys Gly Asn Ile Leu Leu Tyr Met Asp Ile Asp Arg Gly Ser Arg Tyr Thr Leu Gly His Val His Ile Gln Gly Phe Glu Val Leu Pro Lys Arg Leu Ile Glu Lys Gln Ser Gln Val Gly Pro Asn Asp Leu Tyr Cys Pro Asp Lys Ile Trp Asp Gly Ala His Lys Ile Lys Gln Thr Tyr Ala Lys Tyr Gly Tyr Ile Asn Thr Asn Val Asp Val Leu Phe Ile Pro His Ala Thr Arg Pro Ile Tyr Asp Vai Thr Tyr Glu Val Ser Glu Gly Ser Pro Tyr Lys Val Gly Leu Ile Lys Ile Thr Gly Asn Thr His Thr Lys Ser Asp Val Ile Leu His Glu Thr Ser Leu Phe Pro Gly Asp Thr Phe Asn Arg Leu Lys Leu Glu Asp Thr Glu Gln Arg Leu Arg Asn Thr Gly Tyr Phe Gln Ser Val Ser Val Tyr Thr Val Arg Ser Gln Leu Asp Pro Met Gly Asn Ala Asp Gln Tyr Arg Asp Ile Phe Val Glu Val Lys Glu Thr Thr Thr Gly Asn Leu Gly Leu Phe Leu Gly Phe Ser Ser Leu Asp Asn Leu Phe Gly Gly Ile Glu Leu Ser Glu Ser Asn Phe Asp Leu Phe Gly Ala Arg Asn Ile Phe Ser Lys Gly Phe Arg Cys Leu Arg Gly Gly Gly Glu His Leu Phe Leu Lys Ala Asn Phe Gly Asp Lys Val Thr Asp Tyr Thr Leu Lys Trp Thr Lys Pro His Phe Leu Asn Thr Pro Trp Ile Leu Gly Ile Glu Leu Asp Lys Ser Ile Asn Arg Ala Leu Ser Lys Asp Tyr Ala Val Gln Thr Tyr Gly Gly Asn Val Ser Thr Thr Tyr Ile Leu Asn Glu His Leu Lys Tyr Gly Leu Phe Tyr Arg Gly Ser Gln Thr Ser Leu His Glu Lys Arg Lys Phe Leu Leu Gly Pro Asn Ile Asp Ser Asn Lys Gly Phe Val Ser Ala Ala Gly Val Asn Leu Asn Tyr Asp Ser Val Asp Ser Pro Arg Thr Pro Thr Thr Gly Ile Arg Gly Gly Val Thr Phe Glu Val Ser Gly Leu Gly Gly Thr Tyr His Phe Thr Lys Leu Ser Leu Asn Ser Ser Ile Tyr Arg Lys Leu Thr Arg Lys Gly Ile Leu Lys Ile Lys Gly Glu Ala Gln Phe Ile Lys Pro Tyr Ser Asn Thr Thr Ala Glu Gly Asp Pro Val Sex Glu Arg Phe Phe Leu Gly Gly Glu Thr Thr Val Arg Gly Tyr Lys Ser Phe Ile Ile Gly Pro Lys Tyr Ser Ala Thr Glu Pro Gln Gly Gly Leu Ser Ser Leu Leu Ile Ser Glu Glu Phe Gln Tyr Pro Leu Ile Arg Gln Pro Asn Ile Ser Ala Phe Val Phe Leu Asp Ser Gly Phe Val Gly Leu Gln Glu Tyr Lys Ile Ser Leu Lys Asp Leu Arg Ser Ser Ala Gly Phe Gly Leu Arg Phe Asp Val Met Asn Asn Val Pro Val Met Leu Gly Phe Gly Trp Pro Phe Arg Pro Thr Glu Thr Leu Asn Gly Glu Lys Ile Asp Val Ser Gln Arg Phe Phe Phe Ala Leu Gly 770 775 7so Gly Met Phe <210> 3 <211> 3200 <212> DNA
<213> Chlamydia sp.
<220>
<221> CDS
<222> (101)..(3010) <400> 3 gggaactttt attcgtttta ttaagtaaga tcgagtatgg tattgagaaa cgttctcgta 60 aaagatggta ttctagtaat tacagtgaat taaacgacat atg tta ggt ttt ctt 115 Met Leu Gly Phe Leu aaa cgc ttc ttt ggt tcc tct caa gag cgt att cta aaa aaa ttt caa 163 Lys Arg Phe Phe Gly Ser Ser Gln Glu Arg Ile Leu Lys Lys Phe Gln aaa ctt gta gat aaa gtg aac att tat gat gaa atg ctc acg cct tta 211 Lys Leu Val Asp Lys Val Asn Ile Tyr Asp Glu Met Leu Thr Pro Leu tct gat gat gaa ttg cgg aat aag act gcg gaa tta aag cag aga tat 259 Ser Asp Asp Glu Leu Arg Asn Lys Thr Ala Glu Leu Lys Gln Arg Tyr cag aac gga gag tct ctt gat agc atg ctt cct gaa get tat ggt gtc 307 Gln Asn Gly Glu Ser Leu Asp Ser Met Leu Pro Glu Ala Tyr Gly Val gtg aaa aac gtt tgt cga cgt tta gca ggc acc cca gtc gaa gtc tcc 355 Val Lys Asn Val Cys Arg Arg Leu Ala Gly Thr Pro Val Glu Val Ser 70 7s eo es gga tac cat caa aga tgg gat atg gtt cct tat gat gtg cag att cta 403 Gly Tyr His Gln Arg Trp Asp Met Val Pro Tyr Asp Val Gln Ile Leu ggg gcc att get atg cac aag gga ttt att aca gag atg cag acc ggg 451 Gly Ala Ile Ala Met His Lys Gly Phe Ile Thr Glu Met Gln Thr Gly gag ggg aaa aca ctc act gca gtg atg cct ctg tat tta aat get tta 499 Glu Gly Lys Thr Leu Thr Ala Val Met Pro Leu Tyr Leu Asn Ala Leu aca ggc aag cca gtg cat ttg gtt act gtt aac gac tat ctt gca caa 547 Thr Gly Lys Pro Val His Leu Val Thr Val Asn Asp Tyr Leu Ala Gln cga gat tgt gaa tgg gta gga tcg gta ctg cgc tgg tta gga ctt aca 595 Arg Asp Cys Glu Trp Val Gly Ser-Val Leu Arg Trp Leu Gly Leu Thr acg gga gtt ttg gtt tca gga act ctt tta gaa aag cgt aag aaa att 643 Thr Gly Val Leu Val Ser Gly Thr Leu Leu Glu Lys Arg Lys Lys Ile tat caa tgt gat gtt gtc tat ggt aca gca tct gag ttt ggt ttt gat 691 Tyr Gln Cys Asp Val Val Tyr Gly Thr Ala Ser Glu Phe Gly Phe Asp tat ttg aga gat aat tct ata get act cgc ctt gaa gag cag gta ggt 739 Tyr Leu Arg Asp Asn Ser Ile Ala Thr Arg Leu Glu Glu Gln Val Gly aga gga tat tac ttt get atc att gat gaa gtc gac tcg atc tta ata 787 Arg Gly Tyr Tyr Phe Ala Ile Ile Asp Glu Val Asp Ser Ile Leu Ile gat gaa get aga aca ccc tta att atc tca ggt cct gga gaa aaa cat 835 Asp Glu Ala Arg Thr Pro Leu Ile Ile Ser Gly Pro Gly Glu Lys His aat cca gtc tat ttt gag ctt aaa gaa aaa gtc gca agt cta gtg tat 883 Asn Pro Val Tyr Phe Glu Leu Lys Glu Lys Val Ala Ser Leu Val Tyr ttg caa aaa gag ctc tgc agc cgt atc gca tta gaa gca cgt cgc ggc 931 Leu Gln Lys Glu Leu Cys Ser Arg Ile Ala Leu Glu Ala Arg Arg Gly tta gat agc ttt tta gat gtt gat att ctt cct aaa gat aaa aaa gtt 979 Leu Asp Ser Phe Leu Asp Val Asp Ile Leu Pro Lys Asp Lys Lys Val ctt gaa ggc atc tct gaa ttt tgc cgc agc ctt tgg ttg gta agc aaa 1027 Leu Glu Gly Ile Ser Glu Phe Cys Arg Ser Leu Trp Leu Val Ser Lys gga atg cct ttg aat cgt gtg tta cgt cgt gta cgt gag cac cca gat 1075 Gly Met Pro Leu Asn Arg Val Leu Arg Arg Val Arg Glu His Pro Asp ctt cgt get atg atc gat aaa tgg gat gtt tat tat cat get gag cag 1123 Leu Arg Ala Met Ile Asp Lys Trp Asp Val Tyr Tyr His Ala Glu Gln aat aaa gaa gag agc cta gag cgt ctt tca gag ctc tac att att gtt 1171 Asn Lys Glu Glu Ser Leu Glu Arg Leu Ser Glu Leu Tyr Ile Ile Val gat gag cac aat aat gat ttt gag ctt aca gat aaa gga atg cag cag 1219 Asp Glu His Asn Asn Asp Phe Glu Leu Thr Asp Lys Gly Met Gln Gln tgg gtt gag tat get gga ggc tct acc gaa gag ttc gtg atg atg gat 1267 Trp Val Glu Tyr Ala Gly Gly Ser Thr Glu Glu Phe Val Met Met Asp atg ggg cat gag tat get ctt ata gaa aat gat gag acc cta tca cct 1315 Met Gly His Glu Tyr Ala Leu Ile Glu Asn Asp Glu Thr Leu Ser Pro gca gat aag atc aat aaa aaa att gca att tct gaa gaa gac acc tta 1363 Ala Asp Lys Ile Asn Lys Lys Ile.Ala Ile Ser Glu Glu Asp Thr Leu aga aag get cgt get cac gga tta cga cag tta tta cga gcc caa ctt 1411 Arg Lys Ala Arg Ala His Gly Leu Arg Gln Leu Leu Arg Ala Gln Leu ctc atg gag cgt gat gta gat tat att gtc cgc gac gat cag att gtg 1459 Leu Met Glu Arg Asp Val Asp Tyr Ile Val Arg Asp Asp Gln Ile Val att atc gat gaa cat aca gga cgt cct caa cct gga cga cgt ttt tct 1507 Ile Ile Asp Glu His Thr Gly Arg Pro Gln Pro Gly Arg Arg Phe Ser gaa ggc ctc cat caa get atc gaa get aaa gaa cac gtc act atc cgt 1555 Glu Gly Leu His Gln Ala Ile Glu Ala Lys Glu His Val Thr Ile Arg aag gaa tct cag acg ctt get aca gtc acg ttg caa aat ttc ttc cgt 1603 Lys Glu Ser Gln Thr Leu Ala Thr Val Thr Leu Gln Asn Phe Phe Arg cta tat gaa aag ctt gca ggg atg acc gga aca gca att acg gag tct 1651 Leu Tyr Glu Lys Leu Ala Gly Met Thr Gly Thr Ala Ile Thr Glu Ser cga gag ttt aaa gaa att tat aat ctt tat gtc ctc caa gta ccc acg 1699 Arg Glu Phe Lys Glu Ile Tyr Asn Leu Tyr Val Leu Gln Val Pro Thr ttc aag ccc ttg tta cgc ata gat cat aat gat gaa ttt tat atg aca 1747 Phe Lys Pro Leu Leu Arg Ile Asp His Asn Asp Glu Phe Tyr Met Thr gag cgt gag aag tac cac get att gtt aat gag att gcg act att cat 1795 Glu Arg Glu Lys Tyr His Ala Ile Val Asn Glu Ile Ala Thr Ile His ggc aag ggg aac cct att ctt gtt ggt aca gaa tct gta gag gtc tct 1843 Gly Lys Gly Asn Pro Ile Leu Val Gly Thr Glu Ser Val Glu Val Ser gag aag ctg tct cgg att ttg aga cag aat cgg ata gag cat act gta 1891 Glu Lys Leu Ser Arg Ile Leu Arg Gln Asn Arg Ile Glu His Thr Val ttg aat get aag aat cat get caa gaa gca gaa att ata gca gga gca 1939 Leu Asn Ala Lys Asn His Ala Gln Glu Ala Glu Ile Ile Ala Gly Ala ggg aaa tta ggt get gtg act gta get aca aat atg get ggt cga ggc 1987 Gly Lys Leu Gly Ala Val Thr Val Ala Thr Asn Met Ala Gly Arg Gly aca gac atc aaa cta gat aat gaa get gtg atc gta ggc ggt ctc cat 2035 Thr Asp Ile Lys Leu Asp Asn Glu Ala Val Ile Val Gly Gly Leu His gtg atc ggt act aca cgg cat caa tcc cgt cga att gat aga cag ttg 2083 Val Ile Gly Thr Thr Arg His Gln Ser Arg Arg Ile Asp Arg Gln Leu cgt ggg cgt tgt get cgt tta gga gac cct ggc get gcg aaa ttc ttt 2131 Arg Gly Arg Cys Ala Arg Leu Gly Asp Pro Gly Ala Ala Lys Phe Phe tta tct ttt gaa gat cgg ctg atg cga cta ttc gcc tcc cct aaa ttg 2179 Leu Ser Phe Glu Asp Arg Leu Met Arg Leu Phe Ala Ser Pro Lys Leu 6gp 685 690 aat acc ctt atc cgt cat ttt cgt cct cca gaa gga gag gcg atg tcg 2227 Asn Thr Leu Ile Arg His Phe Arg Pro Pro Glu Gly Glu Ala Met Ser gac cct atg ttt aat aga ctc ata gaa aca.gca cag aaa cgt gtc gaa 2275 Asp Pro Met Phe Asn Arg Leu Ile Glu Thr Ala Gln Lys Arg Val Glu ggg aga aac tat act atc cgt aag cat acc tta gag tat gat gat gtc 2323 Gly Arg Asn Tyr Thr Ile Arg Lys His Thr Leu Glu Tyr Asp Asp Val atg aat aag cag aga caa geg ata tac get tte cgc cat gat gtc tta 2371 Met Asn Lys Gln Arg Gln Ala Ile Tyr Ala Phe Arg His Asp Val Leu cat gcg gaa tct gtt ttc gat ctt gca aaa gaa att cta tgc cat gtg 2419 His Ala Glu Ser Val Phe Asp Leu Ala Lys Glu Ile Leu Cys His Val tct ctg atg gta gca tcc tta gtg atg agt gat cgt cag ttc aaa ggg 2467 Ser Leu Met Val Ala Ser Leu Val Met Ser Asp Arg Gln Phe Lys Gly tgg aca ttg cca aat ctt gaa gaa tgg ata acc tca tct ttc cca ata 2515 Trp Thr Leu Pro Asn Leu Glu Glu Trp Ile Thr Ser Ser Phe Pro Ile gcc tta aat ata gaa gaa ctc aga cag ctt aaa gat aca gat tct att 2563 Ala Leu Asn Ile Glu Glu Leu Arg Gln Leu Lys Asp Thr Asp Ser Ile get gaa aag atc get get gaa ttg att caa gag ttt caa gta cgc ttt 2611 Ala Glu Lys Ile Ala Ala Glu Leu Ile Gln Glu Phe Gln Val Arg Phe gat cat atg gta gaa ggg ctc tcc aaa get gga ggg gaa gaa ttg gat 2659 Asp His Met Val Glu Gly Leu Ser Lys Ala Gly Gly Glu Glu Leu Asp gca tct get att tgt aga gat gtc gtt cgg tct gtc atg gtc atg cat 2707 Ala Ser Ala Ile Cys Arg Asp Val Val Arg Ser Val Met Val Met His att gat gag cag tgg cgg att cat ctt gta gat atg gac tta cta cgg 2755 Ile Asp Glu Gln Trp Arg Ile His Leu Val Asp Met Asp Leu Leu Arg g7p 875 880 885 agt gaa gtt ggc cta cgt act gta ggg caa aaa gat cct ttg tta gaa 2803 Ser Glu Val Gly Leu Arg Thr Val Gly Gln Lys Asp Pro Leu Leu Glu ttt aaa cac gag tct ttc tta ctg ttt gag agc ttg att cgt gat att 2851 Phe Lys His Glu Ser Phe Leu Leu Phe Glu Ser Leu Ile Arg Asp Ile cgt att acg att gcg cgg cat ctt ttc cgt ctt gaa ctg acc gta gag 2899 Arg Ile Thr Ile Ala Arg His Leu Phe Arg Leu Glu Leu Thr Val Glu cct aat cct cgt gtc aac aac gtg att cct act gta get acg tct ttt 2947 Pro Asn Pro Arg Val Asn Asn Val Ile Pro Thr Val Ala Thr Ser Phe cat aat aat gtg aat tac ggt ccg tta gag ttg act gta gtt aca gat 2995 His Asn Asn Val Asn Tyr Gly Pro Leu Glu Leu Thr Val Vai Thr Asp tct gaa gat caa gat taagaaataa cctagaagga ggagtccttc taggtatttg 3050 Ser Glu Asp Gln Asp tttcacaggc tacatagttt ttatttttaa ataaatgggg tccattctga ccaaacttgg 3110 tcattatagt agcgaatcca gaattttctg cgattgccgg ctatgaggaa ctgggcatag 3170 tggctcagtg tattgtttgc tatacagaaa 3200 <210> 4 <211> 970 <212> PRT
<213> Chlamydia sp.
<400> 4 Met Leu Gly Phe Leu Lys Arg Phe Phe Gly Ser Ser Gln Glu Arg Ile Leu Lys Lys Phe Gln Lys Leu Val Asp Lys Val Asn Ile Tyr Asp Glu Met Leu Thr Pro Leu Ser Asp Asp Glu Leu Arg Asn Lys Thr Ala Glu Leu Lys Gln Arg Tyr Gln Asn Gly Glu Ser Leu Asp Ser Met Leu Pro Glu Ala Tyr Gly Val Val Lys Asn Val Cys Arg Arg Leu Ala Gly Thr Pro Val Glu Val Ser Gly Tyr His Gln Arg Trp Asp Met Val Pro Tyr Asp Val Gln Ile Leu Gly Ala Ile Ala Met His Lys Gly Phe Ile Thr Glu Met Gln Thr Gly Glu Gly Lys Thr Leu Thr Ala Val Met Pro Leu Tyr Leu Asn Ala Leu Thr Gly Lys Pro Val His Leu Val Thr Val Asn Asp Tyr Leu Ala Gln Arg Asp Cys Glu Trp Val Gly Ser Val Leu Arg Trp Leu Gly Leu Thr Thr Gly Val Leu Val Ser Gly Thr Leu Leu Glu Lys Arg Lys Lys Ile Tyr Gln Cys Asp Val Val Tyr Gly Thr Ala Ser Glu Phe Gly Phe Asp Tyr Leu Arg Asp Asn Ser Ile Ala Thr Arg Leu Glu Glu Gln Val Gly Arg Gly Tyr Tyr Phe Ala Ile Ile Asp Glu Val Asp Ser Ile Leu Ile Asp Glu Ala Arg Thr Pro Leu Ile Ile Ser Gly Pro Gly Glu Lys His Asn Pro Val Tyr Phe Glu Leu Lys Glu Lys Val Ala Ser Leu Val Tyr Leu Gln Lys Glu Leu Cys Ser Arg Ile Ala Leu Glu Ala Arg Arg Gly Leu Asp Ser Phe Leu Asp Val Asp Ile Leu Pro Lys Asp Lys Lys Val Leu Glu Gly Ile Ser Glu Phe Cys Arg Ser Leu WO 00!11183 PCT/IB99/01449 Trp Leu Val Ser Lys Gly Met Pro Leu Asn Arg Val Leu Arg Arg Val Arg Glu His Pro Asp Leu Arg Ala Met Ile Asp Lys Trp Asp Val Tyr Tyr His Ala Glu Gln Asn Lys Glu Glu Ser Leu Glu Arg Leu Ser Glu Leu Tyr Ile Ile Val Asp Glu His Asn~Asn Asp Phe Glu Leu Thr Asp Lys Gly Met Gln Gln Trp Val Glu Tyr Ala Gly Gly Ser Thr Glu Glu Phe Val Met Met Asp Met Gly His Glu Tyr Ala Leu Ile Glu Asn Asp Glu Thr Leu Ser Pro Ala Asp Lys Ile Asn Lys Lys Ile Ala Ile Ser Glu Glu Asp Thr Leu Arg Lys Ala Arg Ala His Gly Leu Arg Gln Leu Leu Arg Ala Gln Leu Leu Met Glu Arg Asp Val Asp Tyr Ile Val Arg Asp Asp Gln Ile Val Ile Ile Asp Glu His Thr Gly Arg Pro Gln Pro Gly Arg Arg Phe Ser Glu Gly Leu His Gln Ala Ile Glu Ala Lys Glu His Val Thr Ile Arg Lys Glu Ser Gln Thr Leu Ala Thr Val Thr Leu Gln Asn Phe Phe Arg Leu Tyr Glu Lys Leu Ala Gly Met Thr Gly Thr Ala Ile Thr Glu Ser Arg Glu Phe Lys Glu Ile Tyr Asn Leu Tyr Val Leu Gln Val Pro Thr Phe Lys Pro Leu Leu Arg Ile Asp His Asn Asp Glu Phe Tyr Met Thr Glu Arg Glu Lys Tyr His Ala Ile Val Asn Glu Ile Ala Thr Ile His Gly Lys Gly Asn Pro Ile Leu Val Gly Thr Glu Ser Val Glu Val Ser Glu Lys Leu Ser Arg Ile Leu Arg Gln Asn Arg Ile Glu His Thr Val Leu Asn Ala Lys Asn His Ala Gln Glu Ala Glu Ile Ile Ala Gly Ala Gly Lys Leu Gly Ala Val Thr Val Ala Thr Asn Met Ala Gly Arg Gly Thr Asp Ile Lys Leu Asp Asn Glu Ala Val Ile Val Gly Gly Leu His Val Ile Gly Thr Thr Arg His Gln Ser Arg Arg Ile Asp Arg Gln Leu Arg Gly Arg Cys Ala Arg Leu Gly Asp Pro Gly Ala Ala Lys Phe Phe Leu Ser Phe Glu Asp Arg Leu Met Arg Leu Phe Ala Ser Pro Lys Leu Asn Thr Leu Ile Arg His Phe Arg Pro Pro Glu Gly Glu Ala Met Ser Asp Pro Met Phe Asn Arg Leu Ile Glu Thr Ala Gln Lys Arg Val Glu Gly Arg Asn Tyr Thr Ile Arg Lys His Thr Leu Glu Tyr Asp Asp Val Met Asn Lys Gln Arg Gln Ala Ile Tyr Ala Phe Arg His Asp Val Leu His Ala Glu Ser Val Phe Asp Leu Ala Lys Glu Ile Leu Cys His Val Ser Leu Met Val~Ala Ser Leu Val Met Ser Asp Arg Gln Phe Lys Gly Trp Thr Leu Pro Asn Leu Glu Glu Trp Ile Thr Ser Ser Phe Pro Ile Ala Leu Asn Ile Glu Glu Leu Arg Gln Leu Lys Asp Thr Asp Sex Ile Ala Glu Lys Ile Ala Ala Glu Leu Ile Gln Glu Phe Gln Val Arg Phe Asp His Met Val Glu Gly Leu Ser Lys Ala Gly Gly Glu Glu Leu Asp Ala Ser Ala Ile Cys Arg Asp Val Val Arg Ser Val Met Val Met His Ile Asp Glu Gln Trp Arg Ile His Leu Val Asp Met Asp Leu Leu Arg Ser'Glu Val Gly Leu Arg Thr Val Gly Gln Lys Asp Pro Leu Leu Glu Phe Lys His Glu Ser Phe Leu Leu Phe Glu Ser Leu Ile Arg Asp Ile Arg Ile Thr Ile Ala Arg His Leu Phe Arg Leu Glu Leu Thr Val Glu Pro Asn Pro Arg Val Asn Asn Val Zle Pro Thr Val Ala Thr Ser Phe His Asn Asn Val Asn Tyr Gly Pro Leu Glu Leu Thr Val Val Thr Asp Ser Glu Asp Gln Asp <210> 5 <211> 1300 <212> DNA
<213> Chlamydia sp.
<220>
<221> CDS
<222> (101)..(1180) <400> 5 gaacaaaacg tttgggcttc actcttgaga aaaacaagag cataaggtaa tttgcagata 60 attccgaata tagtcctctt aatcgaacct tggcaacagc atg ggt gaa aaa aca 115 Met Gly Glu Lys Thr gaa aag gcc acg ccg aag cga ctt aga gat get cgg aaa aaa ggt caa 163 Glu Lys Ala Thr Pro Lys Arg Leu Arg Asp Ala Arg Lys Lys Gly Gln gta gca aaa tct cag gat ttt cct tct gcg gtt acc ttt atc gtc tct 211 Val Ala Lys Ser Gln Asp Phe Pro Ser Ala Val Thr Phe Ile Val Ser atg ttt acg get tte tec cta tcg acc ttt ttt tte aag cat tta ggt 259 Met Phe Thr Ala Phe Ser Leu Ser Thr Phe Phe Phe Lys His Leu Gly ggc ttt ctg gtt tcc atg ctc tca caa get ccc act cgc cat gat cct 30?
Gly Phe Leu Val Ser Met Leu Ser Gln Ala Pro Thr Arg His Asp Pro gta att acc tta ttt tat ttg aag aac tgt ctt atg ctt att tta aca 355 Val. Ile Thr Leu Phe Tyr Leu Lys Asn Cys Leu Met Leu Ile Leu Thr gca tca ctt cec tta ctg gga get gtt get gtt gtt ggc gtc att gta 403 Ala Ser Leu Pro Leu Leu Gly Ala Val Ala Val Val Gly Val Ile Val ggt ttt ctt atc gtt ggt cct aca ttt tct acc gaa gtt ttt aaa cca 451 Gly Phe Leu Ile Val Gly Pro Thr Phe Ser Thr Glu Val Phe Lys Pro gat atc aag aag ttc aac cct att gag aac atc aaa caa aag ttt aaa 499 Asp Ile Lys Lys Phe Asn Pro Ile Glu Asn Ile Lys Gln Lys Phe Lys ata aag act ctc ata gag ctg atc aaa tcg att tta aaa att ttt gga 547 Ile Lys Thr Leu Ile Glu Leu Ile Lys Ser Ile Leu Lys Ile Phe Gly gca gcc tta att tta tac ata acg tta aaa agc aaa gtc tct tta att 595 Ala Ala Leu Ile Leu Tyr Ile Thr Leu Lys Ser Lys Val Ser Leu Ile ata gaa act gca gga gtc tct cct ata att act get caa atc ttc aaa 643 Ile Glu Thr Ala Gly Val Ser Pro Ile Ile Thr Ala Gln Ile Phe Lys gaa att ttt tat aaa gca gta acc tcg ata gga att ttc ttt ttg att 691 Glu Ile Phe Tyr Lys Ala Val Thr Ser Ile Gly Ile Phe Phe Leu Ile gtt gcg att ett gac ctt gtc tat cag cge cac aat tte get aaa gaa 739 Val Ala Ile Leu Asp Leu Val Tyr Gln Arg His Asn Phe Ala Lys Glu tta aag atg gag aag ttt gag gtt aag cag gag ttt aaa gac acg gaa 787 Leu Lys Met Glu Lys Phe Glu Val Lys Gln Glu Phe Lys Asp Thr Glu gga aat cct gag att aaa ggc cgt cgt cga caa att get caa gaa att 835 Gly Asn Pro Glu Ile Lys Gly Arg Arg Arg Gln I1e Ala Gln Glu Ile gcc tat gaa gac tcg tca tca cag gtg aaa cat gca agc acc gta gtc 883 Ala Tyr Glu Asp Ser Ser Ser Gln Val Lys His Ala Ser Thr Val Val tct aat ccc aaa gat att get gtt get att ggc tac atg cct gaa aaa 931 Ser Asn Pro Lys Asp Ile Ala Val Ala Ile Gly Tyr Met Pro Glu Lys tat aaa gca cct tgg atc att gcc atg ggc atc aac tta cga get aaa 979 Tyr Lys Ala Pro Trp Ile Ile Ala Met Gly Ile Asn Leu Arg Ala Lys agg ata ctt gat gaa get gaa aag tac gga att ccc att atg cga aac 1027 Arg Ile Leu Asp Glu Ala Glu Lys Tyr Gly Ile Pro Ile Met Arg Asn gta cct tta gca cat cag ctt ttg gat gaa ggg aag gaa tta aaa ttt 1075 Val Pro Leu Ala His Gln Leu Leu Asp Glu Gly Lys Glu Leu Lys Phe att cca gaa tct act tac gaa get att gga gaa att cta ctc tat atc 1123 Ile Pro Glu Ser Thr Tyr Glu Ala Ile Gly Glu Ile Leu Leu Tyr Ile act tca ctg aat gcg caa aat cct aat aat aaa aat act aac caa cct 1171 Thr Ser Leu Asn Ala Gln Asn Pro Asn Asn Lys Asn Thr Asn Gln Pro gat cat tta taatgaataa gctactcaat ttcgtcagca gaacacttgg 1220 Asp His Leu tggcgatacc gccttaaaca tgatcaataa gtccagcgac ttaatccttg ctctttggat 1280 gatgggcgtt gtcttaatga 1300 WO 00/111$3 PCT/IB99/01449 <210> 6 <211> 360 <212> PRT
<213> Chlamydia sp.
<400> 6 Met Gly Glu Lys Thr Glu Lys Ala Thr Pro Lys Arg Leu Arg Asp Ala Arg Lys Lys Gly Gln Val Ala Lys Ser Gln Asp Phe Pro Ser Ala Val Thr Phe Ile Val Ser Met Phe Thr Ala Phe Ser Leu Ser Thr Phe Phe Phe Lys His Leu Gly Gly Phe Leu Val Ser Met Leu Ser Gln Ala Pro Thr Arg His Asp Pro Val Ile Thr Leu Phe Tyr Leu Lys Asn Cys Leu Met Leu Ile Leu Thr Ala Ser Leu Pro Leu Leu Gly Ala Val Ala Val Val Gly Val Ile Val Gly Phe Leu Ile Val Gly Pro Thr Phe Ser Thr Glu Val Phe Lys Pro Asp Ile Lys Lys Phe Asn Pro Ile Glu Asn Ile Lys Gln Lys Phe Lys Ile Lys Thr Leu Ile Glu Leu Ile Lys Ser Ile Leu Lys Ile Phe Gly Ala Ala Leu Ile Leu Tyr Ile Thr Leu Lys Ser Lys Val Ser Leu Ile Ile Glu Thr Ala Gly Val Ser Pro Ile Ile Thr Ala Gln Ile Phe Lys Glu Ile Phe Tyr Lys Ala Val Thr Ser Ile Gly Ile Phe Phe Leu Ile Val Ala Ile Leu Asp Leu Val Tyr Gln Arg His Asn Phe Ala Lys Glu Leu Lys Met Glu Lys Phe Glu Val Lys Gln Glu Phe Lys Asp Thr Glu Gly Asn Pro Glu Ile Lys Gly Arg Arg Arg Gln Ile Ala Gln Glu Ile Ala Tyr Glu Asp Ser Ser Ser Gln Val Lys His Ala Ser Thr Val Val Ser Asn Pro Lys Asp Ile Ala Val Ala Ile Gly Tyr Met Pro Glu Lys Tyr Lys Ala Pro Trp Ile Ile Ala Met Gly Ile Asn Leu Arg Ala Lys Arg Ile Leu Asp Glu Ala Glu Lys Tyr Gly Ile Pro Ile Met Arg Asn Val Pro Leu Ala His Gln Leu Leu Asp Glu Gly Lys Glu Leu Lys Phe Ile Pro Glu Ser Thr Tyr Glu Ala Ile Gly Glu Ile Leu Leu Tyr Ile Thr Ser Leu Asn Ala Gln Asn Pro Asn Asn Lys Asn Thr Asn Gln Pro Asp His Leu <210> 7 <211> 2350 <212> DNA
<213> Chlamydia sp.
<220>
<221> CDS
<222> (101)..(2230) <400> 7 aatctactta cgaagctatt ggagaaattc tactctatat cacttcactg aatgcgcaaa 60 atcctaataa taaaaatact aaccaacctg atcatttata atg aat aag cta ctc 115 Met Asn Lys Leu Leu aat ttc gtc agc aga aca ctt ggt ggc gat acc gcc tta aac atg atc 163 Asn Phe Val Ser Arg Thr Leu Gly Gly Asp Thr Ala Leu Asn Met Ile aat aag tcc agc gac tta atc ctt get ctt tgg atg atg ggc gtt gtc 211 Asn Lys Ser Ser Asp Leu Ile Leu Ala Leu Trp Met Met Gly Val Val tta atg atc att att cct ttg cct ccg cct atc gtt gac ttg atg atc 259 Leu Met Ile Ile Ile Pro Leu Pro Pro Pro Ile Val Asp Leu Met Ile acc atc aac tta tcg atc tct gta ttc tta ttg atg gtg get ctt tat 307 Thr Ile Asn Leu Ser Ile Ser Val Phe Leu Leu Met Val Ala Leu Tyr att cca agt get ttg cag ctg tct gtt ttt ccc tcg ttg ctc ctc atc 355 Ile Pro Ser Ala Leu Gln Leu Ser Val Phe Pro Ser Leu Leu Leu Ile act acg atg ttc cgc ttg ggg att aat att tcc tct tct cga cag att 403 Thr Thr Met Phe Arg Leu Gly Ile Asn Ile Ser Ser Ser Arg Gln Ile ctc ctt aaa gcg tat gcg agt cat gtc att cag gcc ttc gga gac ttc 451 Leu Leu Lys Ala Tyr Ala Ser His Val Ile Gln Ala Phe Gly~Asp Phe gtg gtt gga ggg aac tat gtg gtc ggg ttc att atc ttc ctc att att 499 Val Val Gly Gly Asn Tyr Val Val Gly Phe Ile Ile Phe Leu Ile Ile aca atc att cag ttt atc gta gta act aag ggt gcc gag cgt gtt gcc 547 Thr Ile Ile Gln Phe Ile Val Val Thr Lys Gly Ala Glu Arg Val Ala gaa gtt get gcc cga ttc cga ttg gat gcg atg cca ggt aaa cag atg 595 Glu Val Ala Ala Arg Phe Arg Leu Asp Ala Met Pro Gly Lys Gln Met gcg att gat gcg gac tta cga get ggt atg att gat gcc aca caa get 643 Ala Ile Asp Ala Asp Leu Arg Ala Gly Met Ile Asp Ala Thr Gln Ala cgt gat aaa agg get caa atc caa aag gaa agt gaa ctc tac gga gcc 691 Arg Asp Lys Arg Ala Gln Ile Gln Lys Glu Ser Glu Leu Tyr Gly Ala atg gac ggt gcc atg aag ttc atc aaa gga gac gtt atc get ggt atc 739 Met Asp Gly Ala Met Lys Phe Ile Lys Gly Asp Val Ile Ala Gly Ile gtt atc tct ttg att aac att gtt ggc ggt ttg acg att ggg gtg get 787 Val Ile Ser Leu Ile Asn Ile Val Gly Gly Leu Thr Ile Gly Val Ala atg cac ggc atg gac ctc get caa gca get cac gtc tac act ctt ctc 835 Met His Gly Met Asp Leu Ala Gln Ala Ala His Val Tyr Thr Leu Leu tcc att gga gat ggt tta gtc tct caa att cct tct ctt ttg att gcg 883 Ser Ile Gly Asp Gly Leu Val Ser Gln Ile Pro Ser Leu Leu Ile Ala ttg aca gcg ggt att gtc acg act cgt gta tcg agt gac aaa aat acg 931 Leu Thr Ala Gly Ile Val Thr Thr Arg Val Ser Ser Asp Lys Asn Thr aac ttg ggt aaa gag att tct act cag ctc gtt aaa gaa cca cga gca 979 Asn Leu Gly Lys Glu Ile Ser Thr Gln Leu Vai Lys Glu Pro Arg Ala cta ctc ctt gca ggt get gca act tta ggg gtt ggt ttc ttc aag ggc 1027 Leu Leu Leu Ala Gly Ala Ala Thr Leu Gly Val Gly Phe Phe Lys Gly ttc cct cta tgg tcc ttc tcc att tta gca tta att ttc gtt gcc tta 1075 Phe Pro Leu Trp Ser Phe Ser Ile Leu Ala Leu Ile Phe Val Ala Leu ggg att ctc cta ctg act aag aaa tca gcg gca gga aaa aaa ggt ggt 1123 Gly Ile Leu Leu Leu Thr Lys Lys Ser Ala Ala Gly Lys Lys Gly Gly ggc tca gga get tca aca acc gta ggg get get ggt gat ggc get get 1171 Gly Ser Gly Ala Ser Thr Thr Val Gly Ala Ala Gly Asp Gly Ala Ala act gtt ggg gat aat ccc gat gac tat tct cta act ctt ccc gta att 1219 Thr Val Gly Asp Asn Pro Asp Asp Tyr Ser Leu Thr Leu Pro Val Ile cta gaa ctt gga aaa gat ctc tct aag ctt atc caa cac aag aca aaa 1267 Leu Glu Leu Gly Lys Asp Leu Ser Lys Leu Ile Gln His Lys Thr Lys tca gga caa agc ttt gtt gat gat atg att cct aaa atg cgg caa get 1315 Ser Gly Gln Ser Phe Val Asp Asp Met Ile Pro Lys Met Arg Gln Ala ctc tat cag gat atc gga atc cga tac cct ggc att cat gtt cgc aca 1363 Leu Tyr Gln Asp Ile Gly Ile Arg Tyr Pro Gly Ile His Val Arg Thr gat tcc cct tct tta gaa gga tac gat tat atg att ctg ctt aat gaa 1411 Asp Ser Pro Ser Leu Glu Gly Tyr Asp Tyr Met Ile Leu Leu Asn Glu gtc cct tat gtg cga gga aaa att cct ccg cac cat gtg tta acc aat 1459 Val Pro Tyr Val Arg Gly Lys Ile Pro Pro His His Val Leu Thr Asn gag gtg gag gac aat ctc agc cgt tat aat cta cct ttc att acc tat 1507 Glu Val Glu Asp Asn Leu Ser Arg Tyr Asn Leu Pro Phe Ile Thr Tyr aag aat get gcg ggt ctt cct tct get tgg gtt agt gaa gat gca aaa 1555 Lys Asn Ala Ala Gly Leu Pro Ser Ala Trp Val Ser Glu Asp Ala Lys 470 475 480 4$5 get att cta gag aag gca gca att aaa tat tgg acg ccg ctc gaa gtg 1603 Ala Ile Leu Glu Lys Ala Ala Ile Lys Tyr Trp Thr Pro Leu Glu Val atc att ctc cat ctt tcg tac ttt ttc cat aaa agc tct caa gag ttt 1651 Ile Ile Leu His Leu Ser Tyr Phe Phe His Lys Ser Ser Gln Glu Phe ttg gga att caa gag gta cgt tct atg atc gaa ttt atg gaa cgt tca 1699 Leu Gly Ile Gln Glu Val Arg Ser Met Ile Glu Phe Met Glu Arg Ser ttc ccg gac tta gtg aag gaa gtc aca agg ctt att cca ttg caa aag 1747 Phe Pro Asp Leu Val Lys Glu Val Thr Arg Leu Ile Pro Leu Gln Lys ctt acg gaa atc ttt aag aga ttg gtt caa gag caa atc tca att aaa 1795 Leu Thr Glu Ile Phe Lys Arg Leu Val Gln Glu Gln Ile Ser Ile Lys gac cta cgt aca atc tta gaa tct ctg agc gag tgg gcg caa act gag 1843 Asp Leu Arg Thr Ile Leu Glu Ser Leu Ser Glu Trp Ala Gln Thr Glu aaa gat aca gtt ttg ctt aca gaa tat gta cgg tct tct tta aag ctt 1891 Lys Asp Thr Val Leu Leu Thr Glu Tyr Val Arg Ser Ser Leu Lys Leu tat atc agc ttc aag ttc tct caa gga caa tca gca att tct gtt tat 1939 Tyr Ile Ser Phe Lys Phe Ser Gln Gly Gln Ser Ala Ile Ser Val Tyr ctc tta gat cca gaa att gaa gag atg att cgt gga gca att aaa cag 1987 Leu Leu Asp Pro Glu Ile Glu Glu Met Ile Arg Gly Ala Ile Lys Gln aca tcg gca ggt tct tac ctt get cta gat cct gat tct gtg aac cta 2035 Thr Ser Ala Gly Ser Tyr Leu Ala Leu Asp Pro Asp Ser Val Asn Leu att tta aaa tct atg agg aat acg atc acg cca aca cct gca gga ggc 2083 Ile Leu Lys Ser Met Arg Asn Thr Ile Thr Pro Thr Pro Ala Gly Gly caa cca cca gta tta ttg aca gca att gat gta aga aga tat gta cga 2131 Gln Pro Pro Val Leu Leu Thr Ala Ile Asp Val Arg Arg Tyr Val Arg aaa tta ata gaa aca gaa ttc cet gac att get gtg att tct tat caa 2179 Lys Leu Ile Glu Thr Glu Phe Pro Asp Ile Ala Val Ile Ser Tyr Gln gaa atc cta cca gaa atc cgc atc cag cct tta gga aga att cag att 2227 Glu Ile Leu Pro Glu Ile Arg Ile Gln Pro Leu Gly Arg Ile Gln Ile ttc taattgatac gttgtcgctc ataggaggca tatggcagca tcaggaggca 2280 Phe caggtggttt aggaggcact cagggtgtca accttgcagc tgtagaagct gcagctgcaa 2340 aagcagatgc 2350 <210> 8 <211> 710 <212> PRT
<213> Chlamydia sp.
<400> 8 Met Asn Lys Leu Leu Asn Phe Val Ser Arg Thr Leu Gly Gly Asp Thr Ala Leu Asn Met Ile Asn Lys Ser Ser Asp Leu Ile Leu Ala Leu Trp Met Met Gly Val Val Leu Met Ile Ile Ile Pro Leu Pro Pro Pro Ile Val Asp Leu Met Ile Thr Ile Asn Leu Ser Ile Ser Val Phe Leu Leu Met Val Ala-Leu Tyr Ile Pro Ser Ala Leu Gln Leu Ser Val Phe Pro Ser Leu Leu Leu Ile Thr Thr Met Phe Arg Leu Gly Ile Asn Ile Ser Ser Ser Arg Gln Ile Leu Leu Lys Ala Tyr Ala Ser His Val Ile Gln Ala Phe Gly Asp Phe Val Val Gly Gly Asn Tyr Val Val Gly Phe Ile Ile Phe Leu Ile Ile Thr Ile Ile Gln Phe Ile Val Val Thr Lys Gly Ala Glu Arg Val Ala Glu Val Ala Ala Arg Phe Arg Leu Asp Ala Met Pro Gly Lys Gln Met Ala Ile Asp Ala Asp Leu Arg Ala Gly Met Ile Asp Ala Thr Gln Ala Arg Asp Lys Arg Ala Gln Ile Gln Lys Glu Ser Glu Leu Tyr Gly Ala Met Asp Gly Ala Met Lys Phe Ile Lys Gly Asp Val Ile Ala Gly Ile Val Ile Ser Leu Ile Asn Ile Val Gly Gly Leu Thr Ile Gly Val Ala Met His Gly Met Asp Leu Ala Gln Ala Ala His Val Tyr Thr Leu Leu Ser Ile Gly Asp Gly Leu Val Ser Gln Ile Pro Ser Leu Leu Ile Ala Leu Thr Ala Gly Ile Val Thr Thr Arg Val Ser Ser Asp Lys Asn Thr Asn Leu Gly Lys Glu Ile Ser Thr Gln Leu Val Lys Glu Pro Arg Ala Leu Leu Leu Ala Gly Ala Ala Thr Leu Gly Val Gly Phe Phe Lys Gly Phe Pro Leu Trp Ser Phe Ser Ile Leu Ala Leu Ile Phe Val Ala Leu Gly Ile Len'Leu~Leu~Thr Lys Lps Ser Ala Ala Gly Lys Lys Gly Gly Gly Ser Gly Ala Ser Thr Thr Val Gly Ala Ala Gly Asp Gly Ala Ala Thr Val Gly Asp Asn Pro Asp Asp Tyr Ser Leu Thr Leu Pro Val Ile Leu Glu Leu Gly Lys Asp Leu Ser Lys Leu Ile Gln His Lys Thr Lys Ser Gly Gln Ser Phe Val Asp Asp Met Ile Pro Lys Met Arg Gln Ala Leu Tyr Gln Asp Ile Gly Ile Arg Tyr Pro Gly Ile His Val Arg Thr Asp Ser Pro Ser Leu Glu Gly Tyr Asp Tyr Met Ile Leu Leu Asn Glu Val Pro Tyr Val Arg Gly Lys Ile Pro Pro His His Val Leu Thr Asn Glu Val Glu Asp Asn Leu Ser Arg Tyr Asn Leu Pro Phe Ile Thr Tyr Lys Asn Ala Ala Gly Leu Pro Ser Ala Trp Val Ser Glu Asp Ala Lys Ala Ile Leu Glu Lys Ala Ala Ile Lys Tyr Trp Thr Pro Leu Glu Val Ile Ile Leu His Leu Ser Tyr Phe Phe His Lys Ser Ser Gln Glu Phe Leu Gly Ile Gln Glu Val Arg Ser Met Ile Glu Phe Met Glu Arg Ser Phe Pro Asp Leu Val Lys Glu Val Thr Arg Leu Ile Pro Leu Gln Lys Leu Thr Glu Ile Phe Lys Arg Leu Val Gln Glu Gln Tle Ser Ile Lys Asp Leu Arg Thr Ile Leu Glu Ser Leu Ser Glu Trp Ala Gln Thr Glu Lys Asp Thr Val Leu Leu Thr Glu Tyr Val Arg Ser Ser Leu Lys Leu Tyr Ile Ser Phe Lys Phe Ser Gln Gly Gln Ser Ala Ile Ser Val Tyr Leu Leu Asp Pro Glu Ile Glu Glu Met Ile Arg Gly Ala Ile Lys Gln Thr Ser Ala Gly Ser Tyr Leu Ala Leu Asp Pro Asp Ser Val Asn Leu Ile Leu Lys Ser Met Arg Asn Thr Ile Thr Pro Thr Pro Ala Gly Gly Gln Pro Pro Val Leu Leu Thr Ala Ile Asp Val Arg Arg Tyr Val Arg Lys Leu Ile Glu Thr Glu Phe Pro Asp Ile Ala Val Ile Ser Tyr Gln Glu Ile Leu Pro Glu Ile Arg Ile Gln Pro Leu Gly Arg Ile Gln Ile Phe <210> 9 <211> 1400 <212> DNA
<213> Chlamydia sp.
<220>
<221> CDS
<222> (101)..(1297) <400> 9 gctgtgattt cttatcaaga aatcctacca gaaatccgca tccagccttt aggaagaatt 60 cagattttct aattgatacg ttgtcgctca taggaggcat atg gca gca tca gga 115 Met Ala Ala Ser Gly ggc aca ggt ggt tta gga ggc act cag ggt gtc aac ctt gca get gta 163_ Gly Thr Gly Gly Leu Gly Gly Thr Gln Gly Val Asn Leu A1a Ala Val gaa get gca get gca aaa gca gat gca gca gaa gtt gta gcc agc caa 211 Glu Ala Ala Ala Ala Lys Ala Asp Ala Ala Glu Val Val Ala Ser Gln gaa ggt tct gag atg aac atg att caa caa tct cag gac ctg aca aat 259 Glu Gly Ser Glu Met Asn Met Ile Gln Gln Ser Gln Asp Leu Thr Asn ccc gca gca gca aca cgc acg aaa aaa aag gaa gag aag ttt caa act 307 Pro Ala Ala Ala Thr Arg Thr Lys Lys Lys Glu Glu Lys Phe Gln Thr cta gaa tct cgg aaa aaa gga gaa get gga aag get gag aaa aaa tct 355 Leu Glu Ser Arg Lys Lys Gly Glu Ala Gly Lys Ala Glu Lys Lys Ser gaa tct aca gaa gag aag cct gac aca gat ctt get gat aag tat get 403 Glu Ser Thr Glu Glu Lys Pro Asp Thr Asp Leu Ala Asp Lys Tyr Ala tct ggg aat tct gaa atc tct ggt caa gaa ctt cgc ggc ctg cgt gat 451 Ser Gly Asn Ser Glu Ile Ser Gly Gln Glu Leu Arg Gly Leu Arg Asp gca ata gga gac gat get tct cca gaa gac att ctt get ctt gta caa 499 Ala Ile Gly Asp Asp Ala Ser Pro Glu Asp Ile Leu Ala Leu Val Gln gag aaa att aaa gac cca get ctg caa tcc aca get ttg gac tac ctg 547 Glu Lys Ile Lys Asp Pro Ala Leu Gln Ser Thr Ala Leu Asp Tyr Leu gtt caa acg act cca ccc tcc caa ggt aaa tta aaa gaa gcg ctt atc 595 Val Gln Thr Thr Pro Pro Ser Gln Gly Lys Leu Lys Glu Ala Leu Ile caa gca agg aat act cat acg gag caa ttc gga cga act get att ggt 643 Gln Ala Arg Asn Thr His Thr Glu Gln Phe Gly Arg Thr Ala Ile Gly gcg aaa aac atc tta ttt gcc tct caa gaa tat gca gac caa ctg aat 691 Ala Lys Asn Ile Leu Phe Ala Ser Gln Glu Tyr Ala Asp Gln Leu Asn gtt tct cct tca ggg ctt cgc tct ttg tac tta gaa gtg act gga gac 739.
Val Ser Pro Ser Gly Leu Arg Ser Leu Tyr Leu Glu Val Thr Gly Asp aca cat acc tgt gat cag cta ctt tct atg ctt caa gac cgc tat acc 787 Thr His Thr Cys Asp Gln Leu Leu Ser Met Leu Gln Asp Arg Tyr Thr tac caa gat atg get att gtc agc tcc ttt cta atg aaa gga atg gea 835 Tyr Gln Asp Met Ala Ile Val Ser $er Phe Leu Met Lys Gly Met Ala aca gaa tta aaa agg cag ggt ccc tac gta ccc agt gcg caa cta caa 883 Thr Glu Leu Lys Arg Gln Gly Pro Tyr Val Pro Ser Ala Gln Leu Gln gtt ctc atg aca gaa act cgt aac ctg caa gca gtt ctt acc tcg tac 931 Val Leu Met Thr Glu Thr Arg Asn Leu Gln Ala Val Leu Thr Ser Tyr gat tac ttt gaa agt cgc gtt cct att tta ctc gat agc tta aaa get 979 Asp Tyr Phe Glu Ser Arg Val Pro Ile Leu Leu Asp Ser Leu Lys Ala gag gga atc caa act ect tct gat cta aac ttt gtg aag gta get gag 1027 Glu Gly Ile Gln Thr Pro Ser Asp Leu Asn Phe Val Lys Val Ala Glu tcc tac cat aaa atc att aac gat aag ttc cca aca gca tct aaa gta 1075 Ser Tyr His Lys Ile Ile Asn Asp Lys Phe Pro Thr Ala Ser Lys Val gaa cga gaa gtc cgc aat ctc ata gga gac gat gtt gat tct gtg acc 1123 Glu Arg Glu Val Arg Asn Leu Ile Gly Asp Asp Val Asp Ser Val Thr ggt gtc ttg aac tta ttc ttt tet get tta cgt caa acg tcg tca egc 1171 Gly Val Leu Asn Leu Phe Phe Ser Ala Leu Arg Gln Thr Ser Ser Arg ctt ttc tct tca gca gac aaa cgt cag caa tta gga get atg att get 1219 Leu Phe Ser Ser Ala Asp Lys Arg Gln Gln Leu Gly Ala Met Ile Ala aat get tta gat get gta aat ata aac aat gaa gat tat cce aaa gca 1267 Asn Ala Leu Asp Ala Val Asn Ile Asn Asn Glu Asp Tyr Pro Lys Ala tca gac ttc cct aaa ccc tat cct tgg tca tgattaaaaa aggattgcca 1317 Ser Asp Phe Pro Lys Pro Tyr Pro Trp Ser tgcaaaacca atacgagcaa ttactagaat ccttagcacc cctattaaat acgacacttg 1377 ctccagataa aaataactct tgt 1400 <210> 10 <211> 399 <212> PRT
<213> Chlamydia sp.
<400> 10 Met Ala Ala Ser Gly Gly Thr Gly Gly Leu Gly Gly Thr Gln Gly Val Asn Leu Ala Ala Val Glu Ala Ala Ala Ala Lys Ala Asp Ala Ala Glu Val Val Ala Ser Gln Glu Gly Ser Glu Met Asn Met Ile Gln Gln Ser Gln Asp Leu Thr Asn Pro Ala Ala Ala Thr Arg Thr Lys Lys Lys Glu Glu Lys Phe Gln Thr Leu Glu Ser Arg Lys Lys Gly Glu Ala Gly Lys Ala Glu Lys Lys Ser Glu Ser Thr Glu Glu Lys Pro Asp Thr Asp Leu Ala Asp Lys Tyr Ala Ser Gly Asn Ser Glu Ile Ser Gly Gln Glu Leu Arg Gly Leu Arg Asp Ala Ile Gly Asp Asp Ala Ser Pro Glu Asp Ile Leu Ala Leu Val Gln Glu Lys Ile Lys Asp Pro Ala Leu Gln Ser Thr Ala Leu Asp Tyr Leu Val Gln Thr Thr Pro Pro Ser Gln Gly Lys Leu Lys Glu Ala Leu Ile Gln Ala Arg Asn Thr His Thr Glu Gln Phe Gly Arg Thr Ala Ile Gly Ala Lys Asn Ile Leu Phe Ala Ser Gln Glu Tyr Ala Asp Gln Leu Asn Val Ser Pro Ser Gly Leu Arg Ser Leu Tyr Leu Glu Val Thr Gly Asp Thr His Thr Cys Asp Gln Leu Leu Ser Met Leu Gln Asp Arg Tyr Thr Tyr Gln Asp Met Ala Ile Val Ser Ser Phe Leu Met Lys Gly Met Ala Thr Glu Leu Lys Arg Gln Gly Pro Tyr Val Pro Ser Ala Gln Leu Gln Val Leu Met Thr Glu Thr Arg Asn Leu Gln Ala Val Leu Thr Ser Tyr Asp Tyr Phe Glu Ser Arg Val Pro Ile Leu Leu Asp Ser Leu Lys Ala Glu Gly Ile Gln Thr Pro Ser Asp Leu Asn Phe Val Lys Val Ala Glu Ser Tyr His Lys Ile Ile Asn Asp Lys Phe Pro Thr Ala Ser Lys Val Glu Arg Glu Val Arg Asn Leu Ile Gly Asp Asp Val Asp Ser Val Thr Gly Val Leu Asn Leu Phe Phe Ser Ala Leu Arg Gln Thr Ser Ser Arg Leu Phe Ser Ser Ala Asp Lys Arg Gln Gln Leu Gly Ala Met Ile Ala Asn Ala Leu Asp Ala Val Asn Ile Asn Asn Glu Asp Tyr Pro Lys Ala Ser Asp Phe Pro Lys Pro Tyr Pro Trp Ser <210> 11 <211> 650 <212> DNA
<213> Chlamydia sp.
<220>
<221> CDS
<222> (101)..(541) <400> 11 gctaatgctt tagatgctgt aaatataaac aatgaagatt atcccaaagc atcagacttc 60 cctaaaccct atccttggtc atgattaaaa aaggattgcc atg caa aac caa tac 115 Met Gln Asn Gln Tyr gag caa tta cta gaa tcc tta gca ccc cta tta aat acg aca ctt get 163 Glu Gln Leu Leu Glu Ser Leu Ala Pro Leu Leu Asn Thr Thr Leu Ala cca gat aaa aat aac tct tgt tta atc cgt ttc agc gat acc cat gtc 211 Pro Asp Lys Asn Asn Ser Cys Leu Ile Arg Phe Ser Asp Thr His Val cct gtg caa ata gaa gaa gat gga aat tcc gga gat ctt gca gta tcg 259 Pro Val Gln Ile Glu Glu Asp Gly Asn Ser Gly Asp Leu Ala Val Ser aca cta cta ggt act ctt cct gaa aac gta ttt cgc gag cgt att ttc 307 Thr Leu Leu Gly Thr Leu Pro Glu Asn Val Phe Arg Glu Arg Ile Phe aaa get get ctc tct gta aat ggc tcg ttc caa tcc agc atc aag gga 355 Lys Ala Ala Leu Ser Val Asn Gly Ser Phe Gln Ser Ser Ile Lys Gly att cta ggc tac ggt gag gtc act caa cag ctc tat ctt tca gat atc 403 Ile Leu Gly Tyr Gly Glu Val Thr Gln Gln Leu Tyr Leu Ser Asp Ile ctg agt atg aac tac cta aat gga gaa aag tta ttc gag tat ctc aag 451 Leu Ser Met Asn Tyr Leu Asn Gly Glu Lys Leu Phe Glu Tyr Leu Lys ctc ttt tct ttg cat get aag att tgg atg gaa tcc cta aga aca ggg 499 Leu Phe Ser Leu His Ala Lys Ile Trp Met Glu Ser Leu Arg Thr Gly aat ctt cct gac ctt cat gtt ttg gga atc tac tac gtc gcg 541 Asn Leu Pro Asp Leu His Val Leu Gly Ile Tyr Tyr Val Ala tgaatgtttt aaaatacaca aaacactcac cctcagcaca tgcttggaaa cttataggaa 601 cctctcctaa acacgggatt tatctcccac tattttcaat acacacaaa 650 <210> 12 <211> 147 <212> PRT
<213> Chlamydia sp.
<400> 12 Met Gln Asn Gln Tyr Glu Gln Leu Leu Glu Ser Leu Ala Pro Leu Leu Asn Thr Thr Leu Ala Pro Asp Lys Asn Asn Ser Cys Leu Ile Arg Phe Ser Asp Thr His Val Pro Val Gln Ile Glu Glu Asp Gly Asn Ser Gly Asp Leu Ala Val Ser Thr Leu Leu Gly Thr Leu Pro Glu Asn Val Phe Arg Glu Arg Ile Phe Lys Ala Ala Leu Ser Val Asn Gly Ser Phe Gln 65 ,70 75 80 Ser Ser Ile Lys Gly Ile Leu Gly Tyr Gly Glu Val Thr Gln Gln Leu Tyr Leu Ser Asp Ile Leu Ser Met Asn Tyr Leu Asn Gly Glu Lys Leu Phe Glu Tyr Leu Lys Leu Phe Ser Leu His Ala Lys Ile Trp Met Glu Ser Leu Arg Thr Gly Asn Leu Pro Asp Leu His Val Leu Gly Ile Tyr Tyr Val Ala <210>13 <211>3050 <212>DNA

<213>Chlamydia sp.

<220>
<221> CDS
<222> (101) . . (2935) <400> 13 aaatgcttct agtctagggg aacttttttc tatttctacg aagggagtgt cttgtctaga 60 actccatagg gagatcgcac gctaaatatg agaatataat atg aag act tca gtt 115 Met Lys Thr Ser Val tct atg ttg ttg gcc ctg ctt tgc tcg ggg get agc tct att gta ctc 163 Ser Met Leu Leu Ala Leu Leu Cys Ser Gly Ala Ser Ser Ile Val Leu cat gcc gca acc act cca cta aat cct gaa gat ggg ttt att ggg gag 211 His Ala Ala Thr Thr Pro Leu Asn Pro Glu Asp Gly Phe Ile Gly Glu ggc aat aca aat act ttt tct ccg aaa tct aca acg gat get gca gga 259 Gly Asn Thr Asn Thr Phe Ser Pro Lys Ser Thr Thr Asp Ala Ala Gly act acc tac tct ctc aca gga gag gtt ctg tat ata gat ccg ggg aaa 307 Thr Thr Tyr Ser Leu Thr Gly Glu Val Leu Tyr Ile Asp Pro Gly Lys ggt ggt tca att aca gga act tgc ttt gta gaa act get ggc gat ctt 355 Gly Gly Ser Ile Thr Gly Thr Cys Phe Val Glu Thr Ala Gly Asp Leu aca ttt tta ggt aat gga aat acc cta aag ttc ctg tcg gta gat gca 403 Thr Phe Leu Gly Asn Gly Asn Thr Leu Lys Phe Leu Ser Val Asp Ala ggt get aat atc gcg gtt get cat gta caa gga agt aag aat tta agc 451 Gly Ala Asn Ile Ala Val Ala His Val Gln Gly Ser Lys Asn Leu Ser ttc aca gat ttc ctt tct ctg gtg atc aca gaa tct cca aaa tcc get 499 Phe Thr Asp Phe Leu Ser Leu Val Ile Thr Glu Ser Pro Lys Ser Ala gtt act aca gga aaa ggt agc cta gtc agt tta ggt gca gtc caa ctg 547 Val Thr Thr Gly Lys Gly Ser Leu Val Ser Leu Gly Ala Val Gln Leu caa gat ata aac act cta gtt ctt aca agc aat gcc tct gtc gaa gat 595 Gln Asp Ile Asn Thr Leu Val Leu Thr Ser Asn Ala Ser Val Glu Asp ggt ggc gtg att aaa gga aac tcc tgc ttg att cag gga atc aaa aat 643 Gly Gly Val Ile Lys Gly Asn Ser Cys Leu Ile Gln Gly Ile Lys Asn agt gcg att ttt gga caa aat aca tct tcg aaa aaa gga ggg gcg atc 691 Sex Ala Ile Phe Gly Gln Asn Thr Ser Ser Lys Lys Gly Gly Ala Ile tcc acg act caa gga ctt acc ata gag aat aac tta ggg acg cta aag 739 Ser Thr Thr Gln Gly Leu Thr Ile Glu Asn Asn Leu Gly Thr Leu Lys ttc aat gaa aac aaa gca gtg acc tca gga ggc gcc tta gat tta gga 787 Phe Asn Glu Asn Lys Ala Val Thr Ser Gly Gly Ala Leu Asp Leu Gly gcc gcg tct aca ttc act gcg aac cat gag ttg ata ttt tca caa aat 835 Ala Ala Ser Thr Phe Thr Ala Asn His Glu Leu Ile Phe Ser Gln Asn aag act tct ggg aat get gca aat ggc gga gcc ata aat tgc tca ggg 883 Lys Thr Ser Gly Asn Ala Ala Asn Gly Gly Ala Ile Asn Cys Ser Gly gac ctt aca ttt act gat aac act tct ttg tta ctt caa gaa aat agc 931 Asp Leu Thr Phe Thr Asp Asn Thr Ser Leu Leu Leu Gln Glu Asn Ser aca atg cag gat ggt gga get ttg tgt agc aca gga acc ata agc att 979 Thr Met Gln Asp Gly Gly Ala Leu Cys Ser Thr Gly Thr Ile Ser Ile acc ggt agt gat tct atc aat gtg ata gga aat act tca gga caa aaa 1027 Thr Gly Ser Asp Ser Ile Asn Val Ile Gly Asn Thr Ser Gly Gln Lys 2g5 300 305 gga gga gcg att tct gca get tct ctc aag att ttg gga ggg cag gga 1075 Gly Gly Ala Ile Ser Ala Ala Ser Leu Lys Ile Leu Gly Gly Gln Gly ggc get ctc ttt tct aat aac gta gtg act cat gcc acc cct cta gga 1123 Gly Ala Leu Phe Ser Asn Asn Val Val Thr His Ala Thr Pro Leu Gly ggt gcc att ttt atc aac aca gga gga tcc ttg cag ctc ttc act caa 1171 Gly Ala Ile Phe Ile Asn Thr Gly Gly Ser Leu Gln Leu Phe Thr Gln gga ggg gat atc gta ttc gag ggg aat cag gtc act aca aca get cca 1219 Gly Gly Asp Ile Val Phe Glu Gly Asn Gln Val Thr Thr Thr Ala Pro aat get acc act aag aga aat gta att cac ctc gag agc acc gcg aag 1267 Asn Ala Thr Thr Lys Arg Asn Val Ile His Leu Glu Ser Thr Ala Lys tgg aeg gga ctt get gca agt caa ggt aac get atc tat ttc tat gat 1315 Trp Thr Gly Leu Ala Ala Ser Gln Gly Asn Ala Ile Tyr Phe Tyr Asp ccc att acc acc aac gat acg gga gca agc gat aac tta cgt atc aat 1363 Pro Ile Thr Thr Asn Asp Thr Gly Ala Ser Asp Asn Leu Arg Ile Asn gag gtc agt gca aat caa aag ctc tcg gga tct ata gta ttt tct gga 1411 Glu Val Ser Ala Asn Gln Lys Leu Ser Gly Ser Ile Val Phe Ser Gly gag aga ttg tcg aca gca gaa get ata get gaa aat ctt act tcg agg 1459 Glu Arg Leu Ser Thr Ala Glu Ala Ile Ala Glu Asn Leu Thr Ser Arg atc aac cag cct gtc act tta gta gag ggg agc tta gta ctt aaa cag 1507 Ile Asn Gln Pro Val Thr Leu Val Glu Gly Ser Leu Val Leu Lys Gln gga gtg acc ttg atc aca caa gga ttc tcg cag gag cca gaa tcc acg 1555 Gly Val Thr Leu Ile Thr Gln Gly Phe Ser Gln Glu Pro Glu Ser Thr ctt ctt ttg gat ctg ggg acc tca tta aaa get tct aea gaa gat att 1603 Leu Leu Leu Asp Leu Gly Thr Ser Leu Lys Ala Ser Thr Glu Asp Ile gtc atc aca aat tta tct ata aat gcc gat acc att tac gga aag aat 1651 Va1 Ile Thr Asn Leu Ser Ile Asn Ala Asp Thr Ile Tyr Gly Lys Asn cct atc aat att gta get tca gca gcg aat aag aac att acc cta aca 1699 Pro Ile Asn Ile Val Ala Ser Ala Ala Asn Lys Asn Ile Thr Leu Thr gga acc tta gca ctt gta aat gca gat gga get ttc tat gag aac cat 1747 Gly Thr Leu Ala Leu Val Asn Ala Asp Gly Ala Phe Tyr Glu Asn His acc ttg caa gac tct caa gac tat agc ttt gta aag tta tct cca gga 1795 Thr Leu Gln Asp Ser Gln Asp Tyr Ser Phe Val Lys Leu Ser Pro Gly gcg gga ggg act ata att act caa gat get tct cag aag cct ctt gaa 1843 Ala Gly Gly Thr Ile Ile Thr Gln Asp Ala Ser Gln Lys Pro Leu Glu gta get cct tct aga cca cat tat ggc tat caa gga cat tgg aat gtg 1891 Val Ala Pro Ser Arg Pro His Tyr Gly Tyr Gln Gly His Trp Asn Val caa gtc atc cca gga acg gga act caa ccg agc cag gca aat tta gaa 1939 Gln Val Ile Pro Gly Thr Gly Thr Gln Pro Ser Gln Ala Asn Leu Glu tgg gtg cgg aca gga tac ctt ccg aat ccc gaa cgg caa gga tct tta 1987 Trp Val Arg Thr Gly Tyr Leu Pro Asn Pro Glu Arg Gln Gly Ser Leu gtt ccc aat agc ctg tgg ggt tct ttt gtt gat cag cgt get atc caa 2035 Val Pro Asn Ser Leu Trp Gly Ser Phe Val Asp Gln Arg Ala Ile Gln gaa atc atg gta aat agt agc caa atc tta tgt cag gaa cgg gga gtc 2083 Glu Ile Met Val Asn Ser Ser Gln Ile Leu Cys Gln Glu Arg Gly Val tgg gga get gga att get aat ttc cta cat aga gat aaa att aat gag 2131 Trp Gly Ala Gly Ile Ala Asn Phe Leu His Arg Asp Lys Ile Asn Glu cac ggc tat cgc cat agc ggt gtc ggt tat ctt gtg gga gtt ggc act 2179 His Gly Tyr Arg His Ser Gly Val Gly Tyr Leu Val Gly Val Gly Thr cat get ttt tct gat get acg ata aat gcg get ttt tgc cag ctc ttc 2227 His Ala Phe Ser Asp Ala Thr Ile Asn Ala Ala Phe Cys Gln Leu Phe agt aga gat aaa gac tac gta gta tcc aaa aat cat gga act agc tac 2275 Ser Arg Asp Lys Asp Tyr Val Val Ser Lys Asn His Gly Thr Ser Tyr 71p 715 720 725 tca ggg gtc gta ttt ctt gag gat acc cta gag ttt aga agt cca cag 2323 Ser Gly Val Val Phe Leu Glu Asp Thr Leu Glu Phe Arg Ser Pro Gln gga ttc tat act gat agc tcc tca gaa get tgc tgt aac caa gtc gtc 2371 Gly Phe Tyr Thr Asp Ser Ser Ser Glu Ala Cys Cys Asn Gln Val Val act ata gat atg cag ttg tct tac agc cat aga aat aat gat atg aaa 2419 Thr Ile Asp Met Gln Leu Ser Tyr Ser His Arg Asn Asn Asp Met Lys acc aaa tac acg aca tat cca gaa get cag gga tct tgg gca aat gat 2467 Thr Lys Tyr Thr Thr Tyr Pro Glu Ala Gln Gly Ser Trp Ala Asn Asp gtt ttt ggt ctt gag ttt gga gcg act aca tac tac tac cct~aac agt 2515 Val Phe Gly Leu Glu Phe Gly Ala Thr Thr Tyr Tyr Tyr Pro Asn Ser act ttt tta ttt gat tac tac tct ccg ttt ctc agg ctg cag tgc acc 2563 Thr Phe Leu Phe Asp Tyr Tyr Ser Pro Phe Leu Arg Leu Gln Cys Thr tat get cac cag gaa gac ttc aaa gag aca gga ggt gag gtt cgt cac 2611 Tyr Ala His Gln Glu Asp Phe Lys Glu Thr Gly Gly Glu Val Arg His ttt act agc gga gat ctt ttc aat tta gca gtt cct att ggc gtg aag 2659 Phe Thr Ser Gly Asp Leu Phe Asn Leu Ala Val Pro Ile Gly Val Lys ttt gag aga ttt tca gac tgt aaa agg gga tct tat gaa ctt acc ctt 2707 Phe Glu Arg Phe Ser Asp Cys Lys Arg Gly Ser Tyr Glu Leu Thr Leu get tat gtt cct gat gtg att cgc aaa gat ccc aag agc acg gca aca 2755 Ala Tyr Vai Pro Asp Val Ile Arg Lys Asp Pro Lys Ser Thr Ala Thr ttg get agt gga get acg tgg agc acc cac gga aac aat ctc tcc aga 2803 Leu Ala Ser Gly Ala Thr Trp Ser Thr His Gly Asn Asn Leu Ser Arg caa gga tta caa ctg cgt tta ggg aac cac tgt ctc ata aat cct gga 2851 Gln Gly Leu Gln Leu Arg Leu Gly Asn His Cys Leu Ile Asn Pro Gly att gag gtg ttc agt cac gga get att gaa ttg cgg gga tcc tct cgt 2899 Ile Glu Val Phe Ser His Gly Ala Ile Glu Leu Arg Gly Ser Ser Arg aat tat aac atc aat ctc ggg ggt aaa tac cga ttt taatagggaa 2945 Asn Tyr Asn Ile Asn Leu Gly Gly Lys Tyr Arg Phe ctgagagttc cttattgtag agagtattta caaggatttt gagacgaaag agatttcttg 3005 ttgtaggtat ctctccacag ggtctagttc aatttctata gttca 3050 <210> 14 <211> 945 <212> PRT
<213> Chlamydia sp.
<400> 14 Met Lys Thr Ser Val Ser Met Leu Leu Ala Leu Leu Cys Ser Gly Ala Ser Ser Ile Val Leu His Ala Ala Thr Thr Pro Leu Asn Pro Glu Asp Gly Phe Ile Gly Glu Gly Asn Thr Asn Thr Phe Ser Pro Lys Ser Thr Thr Asp Ala Ala Gly Thr Thr Tyr Ser Leu Thr Gly Glu Val Leu Tyr Ile Asp Pro Gly Lys Gly Gly Ser Ile Thr Gly Thr Cys Phe Val Glu Thr Ala Gly Asp Leu Thr Phe Leu Gly Asn Gly Asn Thr Leu Lys Phe Leu Ser Val Asp Ala Gly Ala Asn Ile Ala Val Ala His Val Gln Gly Ser Lys Asn Leu Ser Phe Thr Asp Phe Leu Ser Leu Val Ile Thr Glu Ser Pro Lys Ser Ala Val Thr Thr Gly Lys Gly Ser Leu Val Ser Leu Gly Ala Val Gln Leu Gln Asp Ile Asn Thr Leu Val Leu Thr Ser Asn Ala Ser Val Glu Asp Gly Gly Val Ile Lys Gly Asn Ser Cys Leu Ile Gln Gly Ile Lys Asn Ser Ala Ile Phe Gly Gln Asn Thr Ser Ser Lys Lys Gly Gly Ala Ile Ser Thr Thr Gln Gly Leu Thr Ile Glu Asn Asn Leu Gly Thr Leu Lys Phe Asn Glu Asn Lys Ala Val Thr Ser Gly Gly Ala Leu Asp Leu Gly Ala Ala Ser Thr Phe Thr Ala Asn His Glu Leu Ile Phe Ser Gln Asn Lys Thr Ser Gly Asn Ala Ala Asn Gly Gly Ala Ile Asn Cys Ser Gly Asp Leu Thr Phe Thr Asp Asn Thr Ser Leu Leu Leu Gln Glu Asn Ser Thr Met Gln Asp Gly Gly Ala Leu Cys Ser Thr Gly Thr Ile Ser Ile Thr Gly Ser Asp Ser Ile Asn Val Ile Gly Asn Thr Ser Gly Gln Lys Gly Gly Ala Ile Ser Ala Ala Ser Leu Lys Ile Leu Gly Gly Gln Gly Gly Ala Leu Phe Ser Asn Asn Val Val Thr His Ala Thr Pro Leu Gly Gly Ala Ile Phe Ile Asn Thr Gly Gly Ser Leu Gln Leu Phe Thr Gln Gly Gly Asp Ile Val Phe Glu Gly Asn Gln Val Thr Thr Thr Ala Pro Asn Ala Thr Thr Lys Arg Asn Val Ile His Leu Glu Ser Thr Ala Lys Trp Thr Gly Leu Ala Ala Ser Gln Gly Asn Ala Ile Tyr Phe Tyr Asp Pro Ile Thr Thr Asn Asp Thr Gly Ala Ser Asp Asn Leu Arg Ile Asn Glu Val Ser Ala Asn Gln Lys Leu Ser Gly Ser Ile Val Phe Ser Gly Glu Arg Leu Ser Thr Ala Glu Ala Ile Ala Glu Asn Leu Thr Ser Arg Ile Asn Gln Pro Val Thr Leu Val Glu Gly Ser Leu Val Leu Lys Gln Gly Val Thr Leu Ile Thr Gln Gly Phe Ser Gln Glu Pro Glu Ser Thr Leu Leu Leu Asp Leu Gly Thr Ser Leu Lys Ala Ser Thr Glu Asp Ile Val Ile Thr Asn Leu Ser Ile Asn Ala Asp Thr Ile Tyr Gly Lys Asn Pro Ile Asn Ile Val Ala Ser Ala Ala Asn Lys Asn Ile Thr Leu Thr Gly Thr Leu Ala Leu Val Asn Ala Asp Gly Ala Phe Tyr Glu Asn His Thr Leu Gln Asp Ser Gln Asp Tyr Ser Phe Val Lys Leu Ser Pro Gly Ala Gly Gly Thr Ile Ile Thr Gln Asp Ala Ser Gln Lys Pro Leu Glu Val Ala Pro Ser Arg Pro His Tyr Gly Tyr Gln Gly His Trp Asn Val Gln Val Ile Pro Gly Thr Gly Thr Gln Pro Ser Gln Ala Asn Leu Glu Trp Val Arg Thr Gly Tyr Leu Pro Asn Pro Glu Arg Gln Gly Ser Leu Val Pro Asn Ser Leu Trp Gly Ser Phe Val Asp Gln Arg Ala Ile Gln Glu Ile Met Val Asn Ser Ser Gln Ile Leu Cys Gln Glu Arg Gly Val Trp Gly Ala Gly Ile Ala Asn Phe Leu His Arg Asp Lys Ile Asn Glu His Gly Tyr Arg His Ser Gly Val Gly Tyr Leu Val Gly Val Gly Thr His Ala Phe Ser Asp Ala Thr Ile Asn Ala Ala Phe Cys Gln Leu Phe Ser Arg Asp Lys Asp Tyr Val Val Ser Lys Asn His Gly Thr Ser Tyr Ser Gly Val Val Phe Leu Glu Asp Thr Leu Glu Phe Arg Ser Pro Gln Gly Phe Tyr Thr Asp Ser Ser Ser Glu Ala Cys Cys Asn Gln Val Val Thr Ile Asp Met Gln Leu Ser Tyr Ser His Arg Asn Asn Asp Met Lys Thr Lys Tyr Thr Thr Tyr Pro Glu Ala Gln Gly Ser Trp Ala Asn Asp Val Phe Gly Leu Glu Phe Gly Ala Thr Thr Tyr Tyr Tyr Pro Asn Ser Thr Phe Leu Phe Asp Tyr Tyr Ser Pro Phe Leu Arg Leu Gln Cys Thr Tyr Ala His Gln Glu Asp Phe Lys Glu Thr Gly g2p 825 830 Gly Glu Val Arg His Phe Thr Ser Gly Asp Leu Phe Asn Leu Ala Val Pro Ile Gly Val Lys Phe Glu Arg Phe Ser Asp Cys Lys Arg Gly Ser Tyr Glu Leu Thr Leu Ala Tyr Val Pro Asp Val Ile Arg Lys Asp Pro Lys Ser Thr Ala Thr Leu Ala Ser Gly Ala Thr Trp Ser Thr His Gly Asn Asn Leu Ser Arg Gln Gly Leu Gln Leu Arg Leu Gly Asn His Cys Leu Ile Asn Pro Gly Ile Glu Val Phe Ser His Gly Ala Ile Glu Leu ~:,Y", . , ., .a Arg Gly Ser Ser Arg Asn Tyr Asn Ile Asn Leu Gly Gly Lys Tyr Arg Phe <210> 15 <211> 2957 <212> DNA
<213> Chlamydia sp.
<220>
<221> CDS
<222> (101)..(2854) <400> 15 ttgaaaaaaa gagagttacg gcagctgtaa agtttttaat attgctccct ttgttccatt 60 tatgtagcgt tcagactttg cactaaaacg agggtgtcat atg aga tcg tct ttt 115 Met Arg Ser Ser Phe tcc ttg tta tta ata tct tca tct cta gcc ttt cct ctc tta atg agt 163 Ser Leu Leu Leu Ile Ser Ser Ser Leu Ala Phe Pro Leu Leu Met Ser gtt tct gca gat get gcc gat ctc aca tta ggg agt cgt gac agt tat 211 Val Ser Ala Asp Ala Ala Asp Leu Thr Leu Gly Ser Arg Asp Ser Tyr aat ggt gat aca agc acc aca gaa ttt act cct aaa gcg gca act tct 259 Asn Gly Asp Thr Ser Thr Thr Glu Phe Thr Pro Lys Ala Ala Thr Ser gat get agt ggc acg acc tat att ctc gat ggg gat gtc tcg ata agc 307 Asp Ala Ser Gly Thr Thr Tyr Ile Leu Asp Gly Asp Val Ser Ile Ser caa gca ggg aaa caa acg agc tta acc aca agt tgt ttt tct aac act 355 Gln Ala Gly Lys Gln Thr Ser Leu Thr Thr Ser Cys Phe Ser Asn Thr gca gga aat ctt acc ttc tta ggg aac gga ttt tct ctt cat ttt gac 403 Ala Gly Asn Leu Thr Phe Leu Gly Asn Gly Phe Ser Leu His Phe Asp aat att att tcg tct act gtt gca ggt gtt gtt gtt agc aat aca gca 451 Asn Ile Ile Ser Ser Thr Val Ala Gly Val Val Val Ser Asn Thr Ala get tct ggg att acg aaa ttc tca gga ttt tca act ctt cgg atg ctt 499 Ala Ser Gly Ile Thr Lys Phe Ser Gly Phe Ser Thr Leu~rg Met Leu gca get cct agg acc aca ggt aaa gga gcc att aaa att acc gat ggt 547 Ala Ala Pro Arg Thr Thr Gly Lys Gly Ala Ile Lys Ile Thr Asp Gly ctg gtg ttt gag agt ata ggg aat ctt gat ctt aat gaa aat gcc tct 595 Leu Val Phe Glu Ser Ile Gly Asn Leu Asp Leu Asn Glu Asn Ala Ser agt gaa aat ggg gga gcc atc aat acg aag act ttg tct ttg act ggg 693 Ser Glu Asn Gly Gly Ala Ile Asn Thr Lys Thr Leu Ser Leu Thr Gly agt acg cgg ttt gta gcg ttc ctt ggc aat agc tcg tcg caa caa ggg 691 Ser Thr Arg Phe Val Ala Phe Leu Gly Asn Ser Ser Ser Gln Gln Gly gga gcg atc tat get tct ggt gac tct gtg att tct gag aat gca gga 739 Gly Ala Ile Tyr Ala Ser Gly Asp Ser Val Ile Ser Glu Asn Ala Gly atc ttg agc ttc gga aac aac agt gcg aca aca tca gga ggc gcg atc 787 Ile Leu Ser Phe Gly Asn Asn Ser Ala Thr Thr Ser Gly Gly Ala Ile tct get gaa ggg aac ctt gtg atc tcc aat aac caa aat atc ttt ttc 835 Ser Ala Glu Gly Asn Leu Val Ile Ser Asn Asn Gln Asn Ile Phe Phe gat ggc tgc aaa gca act aca aat ggc gga get att gat tgt aac aaa 883 Asp Gly Cys Lys Ala Thr Thr Asn Gly Gly Ala Ile Asp Cys Asn Lys gca ggg gcg aac cca gac cct atc ttg act ctt tca gga aat gag agc 931 Ala Gly Ala Asn Pro Asp Pro Ile Leu Thr Leu Ser Gly Asn Glu Ser ctg cat ttt ctg aat aac aca gca gga aat agt gga ggt gcg att tat 979 Leu His Phe Leu Asn Asn Thr Ala Gly Asn Ser Gly Gly Ala Ile Tyr acc aaa aaa ttg gtg tta tcc tca gga cga gga gga gtg tta ttt tct 1027 Thr Lys Lys Leu Val Leu Ser Ser Gly Arg Gly Gly Val Leu Phe Ser aac aac aaa get gcg aat get act cct aaa gga ggg gca att gcg att 1075 Asn Asn Lys Ala Ala Asn Ala Thr Pro Lys Gly Gly Ala Ile Ala Ile cta gat tct gga gag att agc att tct gca gat ctc ggc aat atc att 1123 Leu Asp Ser Gly Glu Ile Ser Ile Ser Ala Asp Leu Gly Asn Ile Ile ttc gag ggc aat act acg agc act aca gga agt cct gcg agt gtg acc 1171 Phe Glu Gly Asn Thr Thr Ser Thr Thr Gly Ser Pro Ala Ser Val Thr aga aat get ata gat ctt gca tcg aat gca aaa ttt tta aat ctc cga 1219 Arg Asn Ala Ile Asp Leu Ala Ser Asn Ala Lys Phe Leu Asn Leu Arg gcg act cgg gga aat aaa gtt att ttc tat gat cct atc acg agc tca 1267 Ala Thr Arg Gly Asn Lys Val Ile Phe Tyr Asp Pro Ile Thr Ser Ser gga get act gat aag ctc tct ttg aat aaa get gac gca gga tct gga 1315 Gly Ala Thr Asp Lys Leu Ser Leu Asn Lys Ala Asp Ala Gly Ser Gly aat acc tat gaa ggc tac atc gtt ttc tct gga gag aaa ctc tca gaa 1363 Asn Thr Tyr Glu Gly Tyr Ile Val Phe Ser Gly Glu Lys Leu Ser Glu gag gaa ctt aag aaa cct gac aat ctg aag tct aca ttt aca cag get 1411 Glu Glu Leu Lys Lys Pro Asp Asn Leu Lys Ser Thr Phe Thr Gln Ala gta gag ctt get gca ggt gcc tta gta ttg aaa gat gga gtg act gta 1459 Val Glu Leu Ala Ala Gly Ala Leu Val Leu Lys Asp Gly Val Thr Val gtt gca aat act ata acg cag gtc gag gga tcg aaa gtc gtt atg gat 1507 Val Ala Asn Thr Ile Thr Gln Val Glu Gly Ser Lys Val Val Met Asp gga ggg act act ttt gag gca agc get gag ggg gtc act ctc aat ggc 1555 Gly Gly Thr Thr Phe Glu Ala Ser Ala Glu Gly Val Thr Leu Asn Gly cta gcc att aat ata gat tcc tta gat ggg aca aat aaa get atc att 1603 Leu Ala Ile Asn Ile Asp Ser Leu Asp Gly Thr Asn Lys Ala Ile Ile aag gcg acg gca gca agt aag gat gtt gcc tta tca ggg cct atc atg 1651 Lys Ala Thr Ala Ala Ser Lys Asp Val Ala Leu Ser Gly Pro Ile Met ctt gta gat get cag ggg aac tat tat gag cat cat aat ctc agt caa 1699 Leu Val Asp Ala Gln Gly Asn Tyr Tyr Glu His His Asn Leu Ser Gln cag cag gtc ttt cct tta ata gag ctt tct gca caa gga acg atg act 1747 Gln Gln Val Phe Pro Leu Ile Glu Leu Ser Ala Gln Gly Thr Met Thr act aca gat atc ccc gat acc cca att cta aat act acg aat cac tat 1795 Thr Thr Asp Ile Pro Asp Thr Pro Ile Leu Asn Thr Thr Asn His Tyr ggg tat caa ggg aac tgg aat att gtt tgg gtc gac gat gca act gca 1843 Gly Tyr Gln Gly Asn Trp Asn Ile Val Trp Val Asp Asp Ala Thr Ala aaa aca aaa aat get acc tta act tgg act aaa aca gga tac aag ccg 1891 Lys Thr Lys Asn Ala Thr Leu Thr Trp Thr Lys Thr Gly Tyr Lys Pro aat cca gaa cgt cag gga cct ttg gtt cct aat agc ctg tgg ggt tct 1939 Asn Pro Glu Arg Gln Gly Pro Leu Val Pro Asn Ser Leu Trp Gly Ser ttt gtc gat gtc cgc tcc att cag agc ctc atg gac cgg agc aca agt 1987 Phe Val Asp Val Arg Ser Ile Gln Ser Leu Met Asp Arg Ser Thr Ser tcg tta tct tcg tca aca aat ttg tgg gta tca gga atc gcg gac ttt 2035 Ser Leu Ser Ser Ser Thr Asn Leu Trp Val Ser Gly Ile Ala Asp Phe ttg cat gaa gat cag aaa gga aac caa cgt agt tat cgt cat tct agc 2083 Leu His Glu Asp Gln Lys Gly Asn Gln Arg Ser Tyr Arg His Ser Ser gcg ggt tat gca tta gga gga gga ttc ttc acg get tct gaa aat ttc 2131 Ala Gly Tyr Ala Leu Gly Gly Gly Phe Phe Thr Ala Ser Glu Asn Phe WO 00/111$3 PCT/IB99/01449 ttt aat ttt get ttt tgt cag ctt ttt ggc tac gac aag gac cat ctt 2179 Phe Asn Phe Ala Phe Cys Gln Leu Phe Gly Tyr Asp Lys Asp His Leu gtg get aag aac cat acc cat gta tat gca ggg gca atg agt tac cga 2227 Val Ala Lys Asn His Thr His Val Tyr Ala Gly Ala Met Ser Tyr Arg cac ctc gga gag tct aag acc ctc get aag att ttg tca gga aat tct 2275 His Leu Gly Glu Ser Lys Thr Leu Ala Lys Ile Leu Ser Gly Asn Ser gac tcc cta cct ttt gtc ttc aat get cgg ttt get tat ggc cat acc 2323 Asp Ser Leu Pro Phe Val Phe Asn Ala Arg Phe Ala Tyr Gly His Thr gac aat aac atg acc aca aag tac act ggc tat tct cct gtt aag gga 2371 Asp Asn Asn Met Thr Thr Lys Tyr Thr Gly Tyr Ser Pro Val Lys Gly agc tgg gga aat gat gcc ttc ggt ata gaa tgt gga gga get atc ccg 2419 Ser Trp Gly Asn Asp Ala Phe Gly Ile Glu Cys Gly Gly Ala Ile Pro gta gtt get tca gga cgt cgg tct tgg gtg gat acc cac acg cca ttt 2467 Val Val Ala Ser Gly Arg Arg Ser Trp Val Asp Thr His Thr Pro Phe cta aac cta gag atg atc tat gca cat cag aat gac ttt aag gaa aac 2515 Leu Asn Leu Glu Met Ile Tyr Ala His Gln Asn Asp Phe Lys Glu Asn ggc aca gaa ggc cgt tct ttc caa agt gaa gac ctc ttc aat cta gcg 2563 Gly Thr Glu Gly Arg Ser Phe Gln Ser Glu Asp Leu Phe Asn Leu Ala gtt cct gta ggg ata aaa ttt gag aaa ttc tcc gat aag tct acg tat 2611 Val Pro Val Gly Ile Lys Phe Glu Lys Phe Ser Asp Lys Ser Thr Tyr gat ctc tcc ata get tac gtt ccc gat gtg att cgt aat gat cca ggc 2659 Asp Leu Ser Ile Ala Tyr Val Pro Asp Val Ile Arg Asn Asp Pro Gly tgc acg aca act ctt atg gtt tct ggg gat tct tgg tcg aca tgt ggt 2707 Cys Thr Thr Thr Leu Met Val Ser Gly Asp Ser Trp Ser Thr Cys Gly aca agc ttg tct aga caa get ctt ctt gta cgt get gga aat cat cat 2755 Thr Ser Leu Ser Arg Gln Ala Leu Leu Val Arg Ala Gly Asn His His gcc ttt get tca aac ttt gaa gtt ttc agt cag ttt gaa gtc gag ttg 2803 Ala Phe Ala Ser Asn Phe Glu Val Phe Ser Gln Phe Glu Val Glu Leu cga ggt tct tct cgt agc tat get atc gat ctt gga gga aga ttc gga 2851 Arg Gly Ser Ser Arg Ser Tyr Ala Ile Asp Leu Gly Gly Arg Phe Gly ttt taatcctaag ttttccaacg agatagcatc agggtaagcc agggctctat 2904 Phe gtaagagatt tcatagagcc ctctctttgt cttgcttttt gtattttatt ttt 2957 <210> 16 <211> 918 <212> PRT
<213> Chlamydia sp.
<400> 16 Met Arg Ser Ser Phe Ser Leu Leu Leu Ile Ser Ser Ser Leu Ala Phe Pro Leu Leu Met Ser Val Ser Ala Asp Ala Ala Asp Leu Thr Leu Gly Ser Arg Asp Ser Tyr Asn Gly Asp Thr Ser Thr Thr Glu Phe Thr Pro Lys Ala Ala Thr Ser Asp Ala Ser Gly Thr Thr Tyr Ile Leu Asp Gly Asp Val Ser Ile Ser Gln Ala Gly Lys Gln Thr Ser Leu Thr Thr Ser Cys Phe Ser Asn Thr Ala Gly Asn Leu Thr Phe Leu Gly Asn Gly Phe Ser Leu His Phe Asp Asn Ile Ile Ser Ser Thr Val Ala Gly Val Val Val Ser Asn Thr Ala Ala Ser Gly Ile Thr Lys Phe Ser Gly Phe Ser Thr Leu Arg Met Leu Ala Ala Pro Arg Thr Thr Gly Lys Gly Ala Ile Lys Ile Thr Asp Gly Leu Val Phe Glu Ser Ile Gly Asn Leu Asp Leu Asn Glu Asn Ala Ser Ser Glu Asn Gly Gly Ala Ile Asn Thr Lys Thr Leu Ser Leu Thr Gly Ser Thr Arg Phe Val Ala Phe Leu Gly Asn Ser Ser Ser Gln Gln Gly Gly Ala Ile Tyr Ala Ser Gly Asp Ser Val Ile Ser Glu Asn Ala Gly Ile Leu Ser Phe Gly Asn Asn Ser Ala Thr Thr Ser Gly Gly Ala Ile Ser Ala Glu Gly Asn Leu Val Ile Ser Asn Asn Gln Asn Ile Phe Phe Asp Gly Cys Lys Ala Thr Thr Asn Gly Gly Ala Ile Asp Cys Asn Lys Ala Gly Ala Asn Pro Asp Pro Ile Leu Thr Leu Ser Gly Asn Glu Ser Leu His Phe Leu Asn Asn Thr Ala Gly Asn Ser Gly Gly Ala Ile Tyr Thr Lys Lys Leu Val Leu Ser Ser Gly Arg Gly Gly Val Leu Phe Ser Asn Asn Lys Ala Ala Asn Ala Thr Pro Lys Gly Gly Ala Ile Ala Ile Leu Asp Ser Gly Glu Ile Ser Ile Ser Ala Asp Leu Gly Asn Ile Ile Phe Glu Gly Asn Thr Thr Ser Thr Thr Gly Ser Pro Ala Ser Val Thr Arg Asn Ala Ile Asp Leu Ala Ser Asn Ala Lys Phe Leu Asn Leu Arg Ala Thr Arg Gly Asn Lys Val Ile Phe Tyr Asp Pro Ile Thr Ser Ser Gly Ala Thr Asp Lys Leu Ser Leu Asn Lys Ala Asp Ala Gly Ser Gly Asn Thr Tyr Glu Gly Tyr Ile Val Phe Ser Gly Glu Lys Leu Ser Glu Glu Glu Leu Lys Lys Pro Asp Asn Leu Lys Ser Thr Phe Thr Gln Ala Val Glu Leu Ala Ala Gly Ala Leu Val Leu Lys Asp Gly Val Thr Val Val Ala Asn Thr Ile Thr Gln Val Glu Gly Ser Lys Val Val Met Asp Gly Gly Thr Thr Phe Glu Ala Ser Ala Glu Gly Val Thr Leu Asn Gly Leu Ala Ile Asn Ile Asp Ser Leu Asp Gly Thr Asn Lys Ala Ile Ile Lys Ala Thr Ala Ala Ser Lys Asp Val Ala Leu Ser Gly Pro Ile Met Leu Val Asp Ala Gln Gly Asn Tyr Tyr Glu His His Asn Leu Ser Gln Gln Gln Val Phe Pro Leu Ile Glu Leu Ser Ala Gln Gly Thr Met Thr Thr Thr Asp Ile Pro Asp Thr Pro Ile Leu Asn Thr Thr Asn His Tyr Gly Tyr Gln Gly Asn Trp Asn Ile Val Trp Val Asp Asp Ala Thr Ala Lys Thr Lys Asn Ala Thr Leu Thr Trp Thr Lys Thr Gly Tyr Lys Pro Asn Pro Glu Arg Gln Gly Pro Leu Val Pro Asn Ser Leu Trp Gly Ser Phe Val Asp Val Arg Ser Ile Gln Ser Leu Met Asp Arg Ser Thr Ser Ser Leu Ser Ser Ser Thr Asn Leu Trp Val Ser Gly Ile Ala Asp Phe Leu His Glu Asp Gln Lys Gly Asn Gln Arg Ser Tyr Arg His Ser Ser Ala Gly Tyr Ala Leu Gly Gly Gly Phe Phe Thr Ala Ser Glu Asn Phe Phe Asn Phe Ala Phe Cys Gln Leu Phe Gly Tyr Asp Lys Asp His Leu Val Ala Lys Asn His Thr His Val Tyr Ala Gly Ala Met Ser Tyr Arg His Leu Gly Glu Ser Lys Thr Leu Ala Lys Ile Leu Ser Gly Asn Ser Asp Ser Leu Pro Phe Val Phe Asn Ala Arg Phe Ala Tyr Gly His Thr Asp Asn Asn Met Thr Thr Lys Tyr Thr Gly Tyr Ser Pro Val Lys Gly Ser Trp Gly Asn Asp Ala Phe Gly Ile Glu Cys Gly Gly Ala Ile Pro Val Val Ala Ser Gly Arg Arg Ser Trp Val Asp Thr His Thr Pro Phe Leu Asn Leu Glu Met Ile Tyr Ala His Gln Asn Asp Phe Lys Glu Asn Gly Thr Glu Gly Arg Ser Phe Gln Ser Glu Asp Leu Phe Asn Leu Ala Val Pro Val Gly Ile Lys Phe Glu Lys Phe Ser Asp Lys Ser Thr Tyr Asp Leu Ser Ile Ala Tyr Val Pro Asp Val Ile Arg Asn Asp Pro Gly Cys Thr Thr Thr Leu Met Val Ser Gly Asp Ser Trp Ser Thr Cys Gly Thr Ser Leu Ser Arg Gln Ala Leu Leu Val Arg Ala Gly Asn His His Ala Phe Ala Ser Asn Phe Glu Val Phe Ser Gln Phe Glu Val Glu Leu Arg Gly Ser Ser Arg Ser Tyr Ala Ile Asp Leu Gly Gly Arg Phe Gly Phe

Claims (53)

What is claimed is:

1. An isolated polynucleotide selected from the group consisting of:
(a) a polynucleotide having a nucleotide sequence selected from the group consisting of SEQ ID NOS: 1, 3, 5, 7, 9, 11, 13, and 15, and functional fragments thereof;
(b) a polynucleotide encoding a polypeptide having a sequence that is at least 75%
homologous to an amino acid sequence selected from the group consisting of SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, and 16, and functional fragments thereof;
and (c) a polynucleotide capable of hybridizing under stringent conditions to any of the polynucleotides having a sequence further comprising the nucleotide sequences:
SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, and 15, and functional fragments thereof.

2. The polynucleotide of claim 1, wherein the polynucleotide encodes a functional fragment of the polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, and 16.

3. The polynucleotide of claim 1, wherein the polynucleotide of steps (a) and (c) has the nucleotide sequence of SEQ ID NO: 1, and the polynucleotide of steps (b) encodes a polypeptide having a sequence that is at least 75% homologous to the amino acid sequence of SEQ ID NO: 2.

4. The polynucleotide of claim 1, wherein the polynucleotide of steps (a) and (c) has the nucleotide sequence of SEQ ID NO: 3, and the polynucleotide of steps (b) encodes a polypeptide having a sequence that is at least 75% homologous to the amino acid sequence of SEQ ID NO: 4.

5. The polynucleotide of claim 1, wherein the polynucleotide of steps (a) and (c) has the nucleotide sequence of SEQ ID NO: 5, and the polynucleotide of steps (b) encodes a polypeptide having a sequence that is at least 75% homologous to the amino acid sequence of SEQ ID NO: 6.

6. The polynucleotide of claim 1, wherein the polynucleotide of steps (a) and (c) has the nucleotide sequence of SEQ ID NO: 7, and the polynucleotide of steps (b) encodes a polypeptide having a sequence that is at least 75% homologous to the amino acid sequence of SEQ ID NO: 8.

7. The polynucleotide of claim 1, wherein the polynucleotide of steps (a) and (c) has the nucleotide sequence of SEQ ID NO: 9, and the polynucleotide of steps (b) encodes a polypeptide having a sequence that is at least 75% homologous to the amino acid sequence of SEQ ID NO: 10.

8. The polynucleotide of claim 1, wherein the polynucleotide of steps (a) and (c) has the nucleotide sequence of SEQ ID NO: 11, and the polynucleotide of steps (b) encodes a polypeptide having a sequence that is at least 75% homologous to the amino acid sequence of SEQ ID NO: 12.

9. The polynucleotide of claim 1, wherein the polynucleotide of steps (a) and (c) has the nucleotide sequence of SEQ ID NO: 13, and the polynucleotide of steps (b) encodes a polypeptide having a sequence that is at least 75% homologous to the amino acid sequence of SEQ ID NO: 4.

10. The polynucleotide of claim 1, wherein the polynucleotide of steps (a) and (c) has the nucleotide sequence of SEQ ID NO: 15, and the polynucleotide of steps (b) encodes a polypeptide having a sequence that is at least 75% homologous to the amino acid sequence of SEQ ID NO: 16.

11. An isolated polypeptide having an amino acid sequence that is at least 75%
homologous to an amino acid sequence selected from the group consisting of SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, and 16.

12. The polypeptide of claim 11, wherein the polypeptide has the amino acid sequence SEQ ID NO: 2, or a functional fragment thereof.

13. The polypeptide of claim 11, wherein the polypeptide has the amino acid sequence SEQ ID NO: 4, or a functional fragment thereof.

14. The polypeptide of claim 11, wherein the polypeptide has the amino acid sequence SEQ ID NO: 6, or a functional fragment thereof.

15. The polypeptide of claim 11, wherein the polypeptide has the amino acid sequence SEQ ID NO: 8, or a functional fragment thereof.

16. The polypeptide of claim 11, wherein the polypeptide has the amino acid sequence SEQ ID NO: 10, or a functional fragment thereof.

17. The polypeptide of claim 11, wherein the polypeptide has the amino acid sequence SEQ ID NO: 12, or a functional fragment thereof.

18. The polypeptide of claim 11, wherein the polypeptide has the amino acid sequence SEQ ID NO: 14, or a functional fragment thereof.

19. The polypeptide of claim 11, wherein the polypeptide has the amino acid sequence SEQ ID NO: 16, or a functional fragment thereof.

20. The polynucleotide of claim 1, linked to a second nucleotide sequence encoding a fusion polypeptide.

21. The polynucleotide of claim 20, wherein the fusion polypeptide is a heterologous signal peptide.

22. A polypeptide comprising the polypeptide of claim 21 linked to a fusion polypeptide.

23. The polypeptide of claim 22, wherein the fusion polypeptide is a signal peptide.

24. The polypeptide of claim 23, wherein the fusion polypeptide comprises a heterologous polypeptide having adjuvant activity.

25. An expression cassette, comprising the polynucleotide of claim 1 operably linked to a promoter.

26. An expression vector comprising the expression cassette of claim 25.

27. A host cell comprising the expression cassette of claim 26.

28. The host cell of claim 27, wherein said host cell is a prokaryotic cell.

29. The host cell of claim 27, wherein said host cell is a eukaryotic cell.

30. A method for producing a recombinant polypeptide, comprising:
(a) culturing a host cell of claim 27, under conditions that the allow the expression of the polypeptide; and (b) recovering the recombinant polypeptide.

31. The method of claim 30, wherein the recombinant polypeptide is selected from the group consisting of: CPN 100111, CPN 100224,CPN 100230, CPN 100231, CPN 100232, CPN 100235, CPN 100394, and CPN 100395.

32. A vaccine vector comprising the expression cassette of claim 25.

33. The vaccine vector of claim 32, wherein said host mammal is human.

34. The vaccine vector of claim 32, in a pharmaceutically acceptable excipient.

35. A pharmaceutical composition, comprising a immunologically effective amount of the vaccine vector of claim 32.

36. A method for inducing an immune response in a mammal, comprising:
administering to said mammal an immunologically effective amount of the vaccine vector of claim 32, wherein said administration induces an immune response.

37. A pharmaceutical composition, comprising an immunologically effective amount of the polypeptide of claim 1 and pharmaceutically acceptable diluent.

38. The pharmaceutical composition of claim 37, further comprising an adjuvant.

39. The pharmaceutical composition of claim 37, further comprising one or more known Chlamydia antigens.

40. A method for inducing an immune response in a mammal, comprising:
administering to said mammal an immunologically effective amount of the pharmaceutical composition of claim 37, wherein said administration induces an immune response.

41. A polynucleotide probe reagent capable of detecting the presence of Chlamydia in biological material, comprising a polynucleotide that hybridizes to the polynucleotide of claim 1 under stringent conditions.

42. The polynucleotide probe reagent of claim 41, wherein said reagent is a DNA primer.

43. A hybridization method for detecting the presence of Chlamydia in a sample, comprising the steps of:
(a) obtaining polynucleotide from the sample;
(b) hybridizing said obtained polynucleotide with a polynucleotide probe reagent of claim 41 under conditions which allow for the hybridization of said probe and said sample; and (c) detecting said hybridization of said detecting reagent with a polynucleotide in said sample.

44. An amplification method for detecting the presence of Chlamydia in a sample, comprising the steps of:
(a) obtaining polynucleotide from the sample;
(b) amplifying said obtained polynucleotide using one or more polynucleotide probe reagents of claim 41; and (c) detecting said amplified polypeptide.

45. A method for detecting the presence of Chlamydia in a sample comprising the steps of:
(a) contacting said sample with a detecting reagent that binds to a polypeptide to form a complex, the polypeptide being selected from the group consisting of: CPN
100111, CPN 100224, CPN 100230, CPN 100231, CPN 100232, CPN 100235, CPN 100394, and CPN 100395; and (b) detecting said formed complex.

46. The method of claim 45, wherein said detecting reagent is an antibody.

47. The method of claim 46, wherein said antibody is a monoclonal antibody.

48. The method of claim 46, wherein said antibody is a polyclonal antibody.

49. An affinity chromatography method for substantially purifying a Chlamydia antigen comprising the steps of:

(a) contacting a sample containing the Chlamydia antigen with a detecting reagent that binds to a polypeptide to form a complex, the polypeptide being selected frog the group consisting of: CPN 100111, CPN 100224, CPN 100230, CPN 100231, CPN 100232, CPN 100235, CPN 100394, and CPN 100395;
(b) isolating said formed complex;
(c) dissociating said formed complex; and (d) isolating the dissociated Chlamydia antigen.

50. The method of claim 49, wherein said detecting reagent is an antibody.

51. The method of claim 50, wherein said antibody is a monoclonal antibody.

52. The method of claim 50, wherein said antibody is a polyclonal antibody.

53. An antibody that immunospecifically binds a polypeptide of claim 1, or a fragment or derivative of said antibody containing the binding domain thereof.