AU764256B2 - Methods of identifying antigen gene sequences - Google Patents

Description

WO 00/31115 1 PCT/AU99/01035 METHODS OF IDENTIFYING ANTIGEN GENE SEQUENCES The present invention relates to methods of identifying gene sequences of potential vaccine antigens. Also included are gene sequences and the polypeptides encoded by the gene sequences as well as the use of such sequences to induce a protective immune response in animals. Particularly, the invention relates to identifying potential antigen gene sequences of Mycoplasma, preferably Mycoplasma hyopneumoniae.

INTRODUCTION

The success of a vaccine to a pathogen resides in identifying a suitable antigen of a pathogen which is readily accessible to a host immune system.

Once identified, it can form the basis for protection against the pathogen.

In recent years a number of vaccines have been commercialised by Animal Health companies. Many of the vaccines are based on inactivated whole cell bacterins. Although these vaccines provide a reasonable level of efficacy there is considerable scope for improvement. Therefore the presently available vaccines could be improved on by developing vaccines that were not based on whole cells or fractions of whole cells.

Other means of generating vaccines against pathogens includes the use of crude inactivated antigen mixtures of various proteins. However, the problem with these methods is that they present to the host a variety of antigens, none of which may provide suitable overall protection since the proteins most readily accessible to a host immune system are swamped by the other antigens. Moreover, some pathogens are difficult or expensive to grow because of their fastidious requirements. Some pathogens are also harmful to handle. Therefore, a vaccine which does not require the growing of the cells or bacteria and the processing of whole cells or fractions of whole cells would provide a safer, cheaper and possibly more efficacious vaccine.

One way of achieving a cleaner and more specific vaccine is by using recombinant protective antigens. By providing specific antigens, only those antigens readily accessible to the host immune system may be used.

WO 00/31115 PCT/AU99/01035 2 There is the problem of identifying potential antigens. One way is to create a gene library. However, from the library, it is very time consuming to determine those sequences which may include a potential protective antigen.

To efficiently screen through a large number of potential antigen genes for their efficacy in providing some level of protection from disease it is useful to initially clone them as expressing clones. In this way the need for tedious and time consuming clone analysis and subsequent expression subcloning is avoided. The whole process of antigen discovery is speeded up. Applicants now provide a novel method to ensure that the clones initially investigated were expressing recombinant protein.

Previously, expression libraries have been screened for clones expressing all or part of a specific protein by using an antibody specific for or enriched for the protein of interest.

Identifying the DNA sequences that code for the proteins, also makes it possible, using appropriate expression vehicles, to form recombinant DNA molecules and to transform appropriate hosts prokaryotic or eukaryotic hosts) with those recombinant DNA molecules. Culturing of the transformed hosts then permits the hosts to express the DNA sequences and to produce the desired proteins.

Administering the produced and subsequently isolated proteins, active ingredients or combinations thereof by injection), in an amount sufficient to elicit a protective immune response, provides a means for immunising against infections.

One pathogen for which vaccines have been commercialised by Animal Health companies is Mycoplasma hyopneumoniae. This pathogen causes Enzootic pneumonia in pigs. It rarely causes death, but often results in severe morbidity and reduced performance manifesting in significant depression in feed conversion efficiency resulting in reduced weight gain in pigs. The animals show symptoms of coughing and fever and are often prone to secondary infection by opportunistic microorganisms.

Numerous attempts to provide a vaccine against Mycoplasma hyopneumoniae have not been terribly successful. Particularly, the use of heat inactivated, live or extract of Mycoplasma have proven to be ineffective in providing protection. Some vaccines based on inactivated whole cell bacterins have shown some level of efficacy but there is scope for improvement.

Additionally, Mycoplasma hyopneumoniae is difficult and expensive to grow because of its fastidious requirements. Therefore the presently available vaccines could be improved on by developing vaccines that were not based on whole cells or fractions of whole cells.

Accordingly, it is an object of the present invention to overcome or at least alleviate some of the problems of the prior art.

SUMMARY OF THE INVENTION In one aspect of the present invention there is provided a method of identifying expression proteins translated from a nucleotide sequence in an expression vector, said method comprising the use of a small marker tag coexpressed with a protein translated from the nucleotide sequence.

In a preferred embodiment of the present invention there is provided a use of a polyHis tag for identifying expression proteins encoded by a nucleotide sequence in an expression vector wherein said polyHis tag is co-expressed with said protein.

In another aspect of the present invention there is provided a method for identifying expression proteins encoded by a nucleotide sequence from a mixture o: 20 of nucleotide sequences, said method comprising the steps of: providing an expression vector including a small marker tag; oo°introducing a nucleotide sequence from the mixture into the expression vector; and identifying an expression protein expressed by the expression vector by determining the presence of a fusion protein comprising the small marker tag co- S. expressed with the expression protein.

oil In a preferred embodiment the small marker tag is a polyHis tag.

a In a further preferred embodiment, the expression vector is transfected into :a host cell for expression of a marker-fusion protein comprising an expression 30 protein. Preferably the marker fusion protein is a polyHis tag-fusion protein comprising a polyHis tag and an expression protein. Introduction of a population of expression vectors into the host cell may create a genomic or cDNA library which may be screened.

4 In a further preferred embodiment of the method there is included a step of purifying the expression protein using the small marker tag expressed in the fusion protein. Preferably the small marker tag is a polyHis tag.

There is provided in another aspect, a purified recombinant protein identified according to the above method. Preferably the recombinant protein is an expression protein.

In another aspect, there is provided a purified nucleotide sequence which encodes a recombinant protein identified according to the above method.

In another aspect of the present invention there is provided an expression vector including a small marker tag, preferably a polyHis tag for use in identifying expression proteins encoded by a nucleotide sequence.

In yet another aspect of the invention, there is provided a host including an expression vector, said vector including a marker, preferably a polyHis tag for use in identifying an expression protein.

In a further aspect of the present invention, there is provided a method of identifying gene sequences encoding antigenic proteins from a sample of nucleotide sequences, said method comprising the steps of: providing an expression vector including a small marker tag; oooo introducing a nucleotide sequence from the sample into the expression vector; :identifying an antigenic expression protein expressed by the expression i vector by determining the presence of a fusion protein comprising the small marker tag co-expressed with the expression protein translated by the nucleotide sequence; and identifying the antigenic expression protein.

Preferably the small marker tag is a polyHis tag.

In a preferred embodiment, the method further includes determining the gene sequence of the antigenic protein encoded by the expression vector.

In another aspect of the invention there is provided a method of screening 30 a library of nucleotide sequences, said method including the steps of: providing a source of nucleotide sequences; providing an expression vector including a small marker tag; identifying an expression protein expressed by the expression vector by determining the presence of a fusion protein comprising the small marker tag coexpressed with the expression protein translated by the nucleotide sequence; and determining the nucleotide sequence of the expression protein.

Preferably the small marker tag is a polyHis tag.

In another aspect, there is provided a method of screening to identify expression clones in an expression library wherein said method includes: providing a source of nucleotide sequence; providing an expression vector including a small marker tag; introducing the nucleotide sequence into the expression vector; transforming the expression vector into a host to create an expression library; and locating an expression clone by detecting the expression of a fusion protein, comprising the small marker tag, preferably a polyHis tag co-expressed with an expression protein.

In a further aspect of the present invention there is provided a method of identifying a therapeutic antigenic gene sequence encoding a therapeutic antigenic protein of a disease, from a sample of nucleotide sequences, said method comprising the steps of: 20 providing an expression vector including a small marker tag; introducing a nucleotide sequence from the sample into the expression S vector; S°identifying an antigenic expression protein expressed by the expression vector by determining the presence of a fusion protein comprising the small marker tag, co-expressed with the expression protein translated by the nucleotide sequence; inoculating an animal with the expression vector which expresses the oo* antigenic expression protein; challenging the animal with the disease; 30 identifying an expression vector and expression protein that provides a therapeutic effect induced by vaccination; and determining the gene sequence of the antigenic expression protein encoded by the expression vector.

Preferably the small marker tag is a polyHis tag.

6 determining the gene sequence of the antigenic expression protein encoded by the expression vector.

Preferably the small marker tag is a polyHis tag.

In a further preferred embodiment of the invention, there is provided a method of screening a library of nucleotide sequences for a nucleotide sequence encoding polypeptides of Mycoplasma, preferably Mycoplasma hyopneumoniae, said method comprising the steps of: providing a source of nucleotide sequences from a sample of Mycoplasma; providing an expression vector including a small marker tag; introducing a nucleotide sequence into the expression vector; identifying a Mycoplasma expression protein expressed by the expression vector by determining the presence of a fusion protein comprising the small marker tag, preferably a polyHis tag, co-expressed with the expression protein translated by a nucleotide sequence; and determining the nucleotide sequence of the expression protein of Mycoplasma, preferably Mycoplasma hyopneumoniae.

Preferably the small marker tag is a polyHis tag.

In a preferred embodiment, the gene sequence encodes an antigenic polypeptide of Mycoplasma, preferably Mycoplasma hyopneumoniae.

20 In yet another aspect of the present invention, there is provided a nucleotide sequence, mutant, analogue, derivative or functionally active fragment thereof encoding a polypeptide of Mycoplasma, preferably Mycoplasma hyopneumoniae, said nucleotide sequence identified by the methods described above. Preferably the nucleotide sequence comprises a sequence according to any one of Figures 1 to 42, and mutants, analogues derivatives or functionally active fragments thereof. More specifically, the nucleotide sequence is according to SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ S. ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ 30 ID NO:31, SEQ ID NO:33, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:41, SEQ ID NO:43, SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53, SEQ ID WO 00/31115 PCT/AU99/01035 7 SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:61, SEQ ID NO:63, SEQ ID SEQ ID NO:67, SEQ ID NO:69, SEQ ID NO:71, SEQ ID NO:73, SEQ ID SEQ ID NO:77, SEQ ID NO:79, SEQ ID NO:81 or SEQ ID NO:83 or a mutant, analogue, derivatives or functionally active fragments thereof.

In an even further aspect of the present invention, there is provided a polypeptide encoded by the nucleotide sequence mutant, analogue, derivatives or functionally active fragments thereof encoding a polypeptide of Mycoplasma, preferably Mycoplasma hyopneumoniae, said nucleotide sequence identified by the methods described above. Preferably the polypeptide is encoded by a nucleotide sequence encoded by a sequence according to any one of Figures 1 to 42, mutant, analogues, derivatives or functionally active fragment thereof.

More specifically the polypeptide has an amino acid sequence according to SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID SEQ ID NO:32, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:38, SEQ ID SEQ ID NO:42, SEQ ID NO:44, SEQ ID NO:46, SEQ ID NO:48, SEQ ID SEQ ID NO:52, SEQ ID NO:54, SEQ ID NO:56, SEQ ID NO:58, SEQ ID SEQ ID NO 62, SEQ ID NO:64, SEQ ID NO:66, SEQ ID NO:68, SEQ ID SEQ ID NO:72, SEQ ID NO 74, SEQ ID NO: 76, SEQ ID NO:78, SEQ ID SEQ ID NO:82 or SEQ ID NO:84 or a mutant, analogue, derivatives or functionally active fragments thereof.

In a further aspect of the present invention, there is included compositions comprising the polypeptides or nucleotide sequences described above. Furthermore in other aspects, there is included pharmaceutical compositions and vaccine compositions of the polypeptide or nucleotide sequences described above.

Other aspects of the invention include monoclonal antibodies generated against the polypeptides.

Also included as a further aspect of the invention is a method of treating Mycoplasma infection, said method including administering an effective amount of an antigen polypeptide of Mycoplasma encoded by a nucleotide sequence or nucleotide sequence encoding the polypeptide identified by the methods above.

WO 00/31115 PCT/AU99/01035 8 Preferably, the antigen polypeptide is encoded by a nucleotide sequence according to any one of Figures 1 to 42 or a mutant, analogue, derivative or functionally active fragment thereof. More specifically the nucleotide sequence is according to SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:41, SEQ ID NO:43, SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53, SEQ ID NO:55, SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:61, SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:67, SEQ ID NO:69, SEQ ID NO:71, SEQ ID NO:73, SEQ ID NO:75, SEQ ID NO:77, SEQ ID NO:79, SEQ ID NO:81 or SEQ ID NO:83 or a mutant, analogue, derivatives or functionally active fragments thereof.

Throughout the description and claims of this specification, the word "comprise" and variations of the word, such as "comprising" and "comprises", is not intended to exclude other additives, components, integers or steps.

BRIEF DESCRIPTION OF FIGURES Figure 1 shows the nucleotide (SEQ ID NO:1) and amino acid sequence (SEQ ID NO:2) of Mycoplasma represented by clone pAD612.

Figure 2 shows the nucleotide (SEQ ID NO:3) and amino acid sequence (SEQ ID NO:4) of Mycoplasma represented by clone pAD633.

'Figure 3 shows the nucleotide (SEQ ID NO:5) and amino acid sequence (SEQ:ID NO:6) of Mycoplasma represented by clone pAD639.

Figure 4 shows the nucleotide (SEQ ID NO:7) and amino acid sequence (SEQ ID NO:8) of Mycoplasma represented by clone pAD640.

Figure 5 shows the nucleotide (SEQ ID NO:9) and amino acid sequence (SEQ ID NO:10) of Mycoplasma represented by clones pAD641 and pAD1033.

Figure 6 shows the nucleotide (SEQ ID NO:11) and amino acid sequence (SEQ ID NO:12) of Mycoplasma represented by clone pAD653.

Figure 7 shows the nucleotide (SEQ ID NO:13) and amino acid sequence (SEQ ID NO:14) of Mycoplasma represented by clones pAD657 and pAD964.

WO 00/31115 PCT/AU99/01 035 9 Figure 8 shows the nucleotide (SEQ ID NO:15) and amino acid sequence (SEQ ID NO:16) of Mycoplasma represented by clones pAD659 and pAD910.

Figure 9 shows the nucleotide (SEQ ID NO:17) and amino acid sequence (SEQ ID NO:18) of Mycoplasma represented by clone pAD662.

Figure 10 shows the nucleotide (SEQ ID NO:19) and amino acid sequence (SEQ ID NO:20) of Mycoplasma represented by clone pAD681.

Figure 11 shows the nucleotide (SEQ ID NO:21) and amino acid sequence (SEQ ID NO:22) of Mycoplasma represented by clone pAD700.

Figure 12 shows the nucleotide (SEQ ID NO:23) and amino acid sequence (SEQ ID NO:24) of Mycoplasma represented by clone pAD711.

Figure 13 shows the nucleotide (SEQ ID NO:25) and amino acid sequence (SEQ ID NO:26) of Mycoplasma represented by clone pAD721.

Figure 14 shows the nucleotide (SEQ ID NO:27) and amino acid sequence (SEQ ID NO:28) of Mycoplasma represented by clone pAD727.

Figure 15 shows the nucleotide (SEQ ID NO:29) and amino acid sequence (SEQ ID NO:30) of Mycoplasma represented by clone pAD742.

Figure 16 shows the nucleotide (SEQ ID NO:31) and amino acid sequence (SEQ ID NO:32) of Mycoplasma represented by clone pAD760.

Figure 17 shows the nucleotide (SEQ ID NO:33) and amino acid sequence (SEQ ID NO:34) of Mycoplasma represented by clones pAD774.

Figure 18 shows the nucleotide (SEQ ID NO:35) and amino acid sequence (SEQ ID NO:36) of Mycoplasma represented by clone pAD784.

Figure 19 shows the nucleotide (SEQ ID NO:37) and amino acid sequence (SEQ ID NO:38) of Mycoplasma represented by clone pAD789.

Figure 20 shows the nucleotide (SEQ ID NO:39) and amino acid sequence (SEQ ID NO:40) of Mycoplasma represented by clones pAD908, pAD981, pAD1013 and pAD1049.

Figure 21 shows the nucleotide (SEQ ID NO:41) and amino acid sequence (SEQ ID NO:42) of Mycoplasma represented by clones pAD913.

Figure 22 shows the nucleotide (SEQ ID NO:43) and amino acid sequence (SEQ ID NO:44) of Mycoplasma represented by clone pAD920.

Figure 23 shows the nucleotide (SEQ ID NO:45) and amino acid sequence (SEQ ID NO:46) of Mycoplasma represented by clone pAD922.

WO 00/31115 Figure 24 shows the nucleotide (SEQ ID NO:47) and amino (SEQ ID NO:48) of Mycoplasma represented by clones pAD923 Figure 25 shows the nucleotide (SEQ ID NO:49) and amino (SEQ ID NO:50) of Mycoplasma represented by clone pAD950.

Figure 26 shows the nucleotide (SEQ ID NO:51) and amino (SEQ ID NO:52) of Mycoplasma represented by clone pAD951.

Figure 27 shows the nucleotide (SEQ ID NO:53) and amino (SEQ ID NO:54) of Mycoplasma represented by clone pAD977.

Figure 28 shows the nucleotide (SEQ ID NO:55) and amino (SEQ ID NO:56) of Mycoplasma represented by clone pAD983.

Figure 29 shows the nucleotide (SEQ ID NO:57) and amino (SEQ ID NO:58) of Mycoplasma represented by clone pAD984.

Figure 30 shows the nucleotide (SEQ ID NO:59) and amino PCT/AU99/01035 acid sequence and pAD925.

acid sequence acid sequence acid sequence acid sequence acid sequence acid sequence (SEQ ID NO:60) of Mycoplasma sequence 30 represented by clone pAD994.

Figure 31 shows the nucleotide (SEQ ID NO:61) and amino acid sequence (SEQ ID NO:62) of Mycoplasma represented by clone pAD1005.

Figure 32 shows the nucleotide (SEQ ID NO:63) and amino acid sequence (SEQ ID NO:64) of Mycoplasma represented by clone pAD1016.

Figure 33 shows the nucleotide (SEQ ID NO:65) and amino acid sequence (SEQ ID NO:66) of Mycoplasma represented by clone pAD1020.

Figure 34 shows the nucleotide (SEQ ID NO:67) and amino acid sequence (SEQ ID NO:68) of Mycoplasma represented by clone pAD1027.

Figure 35 shows the nucleotide (SEQ ID NO:69) and amino acid sequence (SEQ ID NO:70) of Mycoplasma represented by clone pAD1037.

Figure 36 shows the nucleotide (SEQ ID NO:71) and amino acid sequence (SEQ ID NO:72) of Mycoplasma represented by clone pAD1038.

Figure 37 shows the nucleotide (SEQ ID NO:73) and amino acid sequence (SEQ ID NO:74) of Mycoplasma represented by clones pAD1040.

Figure 38 shows the nucleotide (SEQ ID NO:75) and amino acid sequence (SEQ ID NO:76) of Mycoplasma represented by clone pAD702.

Figure 39 shows the nucleotide (SEQ ID NO:77) and amino acid sequence (SEQ ID NO:78) of Mycoplasma represented by clone pAD763.

WO 00/31115 PCT/AU99/01035 11 Figure 40 shows the nucleotide (SEQ ID NO:79) and amino acid sequence (SEQ ID NO:80) of Mycoplasma represented by clone pAD766.

Figure 41 shows the nucleotide (SEQ ID NO:81) and amino acid sequence (SEQ ID NO:82) of Mycoplasma represented by clone pAD957.

Figure 42 shows the nucleotide (SEQ ID NO:83) and amino acid sequence (SEQ ID NO:84) of Mycoplasma represented by clone pAD996.

DETAILED DESCRIPTION OF THE INVENTION In one aspect of the present invention there is provided a method of identifying expression proteins translated from a nucleotide sequence in an expression vector, said method comprising the use of a marker co-expressed with a protein translated from the nucleotide sequence.

In a preferred embodiment, the marker is any marker which can be detected. Detection may be by a monoclonal antibody or specific polyclonal antiserum. Preferably the marker is small, preferably less than 30 amino acids.

Most preferably, the marker is a polyHis tag. Although the tag may also be any one of the following such as: the FLAG tag, an 8 amino acid tag sequence which is available in plasmid vectors from Sigma-Aldrich Inc. USA, the I-SPY epitope 6 amino acid tag available in vectors from Amrad Corporation Ltd., Australia, the Hemagglutinin epitope 9 amino acid tag (Clontech Laboratories Inc.

USA); and the c-Myc epitope 12 amino acid tag (Clontech Laboratories Inc. USA).

When the marker is a polyHis tag, the number of histidine residues is preferably in the range of 4 to 8 histidine residues, most preferably, there are 6 histidine residues. At least four amino acid residues may precede the polyHis tag providing a total of at least 10 amino acids providing there are at least 4 to 8 contiguous histidine residues.

To efficiently screen through a large number of potential antigen genes for their efficacy in providing some level of protection from disease it is useful to initially clone them as expressing clones. In this way the need for tedious and time consuming clone analysis and subsequent expression subcloning is avoided. The whole process of antigen discovery is speeded up. Applicants WO 00/31115 PCT/AU99/01035 12 provide a novel method to ensure that the clones initially investigated were expressing recombinant protein.

This is particularly the case when dealing with a gene library. Within any random library, there are a majority of clones which are in the incorrect orientation for expression or in the wrong reading frame to express a protein.

The use of a marker, preferably a polyHis tag in an expression vector allows for the efficient identification of expressing clones which express a marker, preferably a polyHis tag fusion protein.

In clones that are not producing a recombinant marker-fusion protein or preferably a polyHis fusion protein, or only a short recombinant protein, the small marker or polyHis protein that is encoded is rapidly degraded within the host cell and any marker or polyHis protein that remains binds very weakly to membranes used for colony screening. Thus non-productive clones give a negative result in colony screening whereas clones producing a longer tagged recombinant protein can give a positive signal. Thus a general genomic expression library can be reduced significantly in complexity and therefore allow faster and more efficient screening and subsequent utilisation of desired clones.

Such clones expressing the proteins are deemed as potential protective antigens and because of the strategy used to construct and screen the clones the originally isolated clones could be immediately used in vaccine trials, both in the form of purified plasmid DNA for genetic vaccination, or as a purified marker-fusion recombinant protein, preferably a polyHis purified recombinant fusion protein.

Where the marker is a polyHis tag region, it may be derived from any source that contains the region. For instance, the regions of the vectors pQE3, pQE31 and pQE32 (Qiagen Pty. Ltd. Australia) encoding the polyHis tag may be used. Other sources of polyHis tag may be obtained from Novagen Inc., USA, (pET-1.9-33) or by synthetically making a polyHis tag.

In another aspect of the present invention there is provided a method for identifying expression proteins translated from a nucleotide sequence from a mixture of nucleotide sequences, said method comprising the steps of: providing an expression vector including a marker; WO 00/31115 PCT/AU99/01035 13 introducing a nucleotide sequence from the mixture into the expression vector; and identifying an expression protein expressed by the expression vector by determining the presence of a fusion protein comprising the marker coexpressed with the expression protein.

Preferably, the marker is a polyHis tag. The following specific examples are directed to a polyHis tag marker and polyHis tag fusion proteins. However, any marker which can be detected and co-expressed with the protein in an expressing clone may be used. Accordingly, the present invention is not restricted to polyHis tags. This marker is illustrative only and should not be taken in any way as a restriction on the generality of the invention described.

As described above, identification of expression proteins, particularly from clones derived from a gene library, can expedite the identification of nucleotide sequences encoding potential antigens to pathogens.

The mixture of nucleotide sequences may be derived from any source such as from a tissue sample, microorganism, cell or any component which contains nucleic acid. Preferably the nucleic acid is genomic, mitochondrial, recombinant or mRNA. A sample of DNA may be firstly subjected to any means which fragments the DNA such as by sonication or by enzyme digestion such as restriction enzyme digestion. Alternatively, the nucleotide sequences may derive from PCR of a portion of a DNA sequence described above. These sequences may be open reading frames (ORF). The expression vector preferably includes a polyHis tag. The polyHis tag may be derived from any source harbouring the polyHis tag as discussed above.

Preferably the polyHis tag may be derived from standard vectors such as pQE31 and pQE32 (Qiagen Pty. Ltd,, Australia). These vectors may be encode a T5 promoter, Lac operator and polyHis tag region. This region may cloned into an expression vector which is capable of expressing proteins encoded by the nucleotide sequences. Alternatively, the polyHis tag may be made synthetically.

Preferably the marker or preferably a polyHis tag region is cloned into a eukaryotic expression vector. Most preferably, the vector is pCI (Promega Corporation, Australia) and preferably creates clones pCl30, pCI31, and pCI32.

WO 00/31115 PCT/AU99/01035 14 This series of vectors allows the cloning of fragments downstream of the polyHis tag region in all three reading frames. The cytomegalovirus (CMV) immediate-early enhancer/promoter, chimeric intron, and SV40 late polyadenylation signal in these exemplified clones allow the vectors to be used as eukaryotic expression vectors in cell culture systems and genetic immunisations using purified plasmid DNA. The T5 promoter allows expression of cloned inserts in prokaryotes. The construction of these vectors is significant because they enable the screening of a library to identify a large number of clones that are expressing recombinant proteins.

The nucleotide sequence(s) from the mixture such as from a digestion of genomic DNA may be introduced or cloned into the expression vector by any means known to the skilled addressee. The expression vector may be used to express protein in a host cell.

In a preferred embodiment, the expression vector is transfected into a host cell for expression of the expression protein of polyHis fusion protein comprising a polyHis tag and an expression protein. Introduction of the expression vector into the host cell may create a genomic or cDNA library.

The host cell may be any cell competent to receive the expression vector. Preferably the cell is prokaryotic. Most preferably it is Escherichia coli.

Identification of the expression protein (and consequently any clones or host cells) expressing the protein can be determined by the presence of the polyHis tag on a polyHis tag fusion protein.

The vectors in the present invention allow the general screening for all expressing clones on the basis of the polyHis tag.

The polyHis tag may be identified by anti-polyHis antibodies or with metal-chelate conjugated probes such as nickel resin wherein the polyHis signals are developed by addition of a chromogenic substrate.

In clones that are not producing a recombinant polyHis fusion protein, or only a short recombinant protein, the small polyHis protein that is encoded is rapidly degraded within the host cell and any polyHis protein that remains binds very weakly to membranes used for colony screening. Thus non-productive clones give a negative result in polyHis colony screening whereas clones producing a longer polyHis tagged recombinant protein can give a positive WO 00/31115 PCT/AU99/01035 signal. Thus a general genomic expression library can be reduced significantly in complexity and therefore allow faster and more efficient screening and subsequent utilisation of desired clones.

In a further preferred embodiment of the method there is included a step of purifying the expression protein using the polyHis tag expressed in the fusion protein.

Once an expression protein expressed by an expression clone is identified using the polyHis tag, then this tag may also be used to purify any expression proteins. Any purification method may be used which is known by the skilled addressee. Affinity columns in affinity chromatography with antipolyHis antibodies may be used. Generally, the expressed protein, identified by virtue of the polyHis tag, may be isolated by any means which relies on utilising the polyHis tag.

There is provided in another aspect, a purified recombinant protein identified according to the above method.

Once the protein is isolated, it may be used in any number of ways including vaccine trials in the form of purified proteins. The proteins can be immediately identified as potential antigens. Preferably, they will be protective antigens.

In another aspect, there is provided a purified nucleotide sequence which encodes recombinant protein identified according to the above method.

The proteins identified by the polyHis method described above are expressed by discrete nucleotide sequences, preferably recombinant sequences. By identifying the protein, a clone expressing the protein may be identified which harbours the nucleotide sequence.

These clones may provide purified plasmid DNA, particularly the plasmid DNA may represent a potential protective antigen.

In another aspect of the present invention there is provided an expression vector or a recombinant vector including a marker, preferably a polyHis tag for use in identifying expression proteins encoded by a nucleotide sequence.

The expression vector may be any vector which produces expression of an expression protein. It may be a plasmid or cosmid. It may be introduced WO 00/31115 PCT/AU99/01035 16 into clones for further expansion of the expression protein for the purposes of obtaining the protein for vaccination or identification purposes.

In yet another aspect of the invention, there is provided a host including an expression vector, said vector including a marker, preferably a polyHis tag for use in identifying an expression protein encoded by a nucleotide sequence.

The host cell may be any competent cell which can harbour an expression vector. Preferably, the host cell is prokaryotic or eukaryotic.

In a further aspect of the present invention, there is provided a method of identifying gene sequences encoding antigenic proteins from a sample of nucleotide sequences, said method comprising the steps of: providing an expression vector including a marker, preferably a polyHis tag; introducing a nucleotide sequence from the sample into the expression vector; identifying an antigenic expression protein expressed by the expression vector by determining the presence of a fusion protein comprising a marker, coexpressed with the expression protein translated by the nucleotide sequence; identifying the antigenic protein; and preferably determining the nucleotide sequence of the antigenic protein gene encoded by the expression vector.

Identification of an antigenic protein may include identifying and preferably isolating a fusion protein having a polyHis tag and determining whether the fusion protein can elicit an immune response which is not generated by virtue of the polyHis tag. Determination of an immune response may be conducted by standard in vitro immunological tests using antisera from immune animals conducting ELISA's or antibody/antigen reactions. They may also be tested in vivo in protection trials in animals against further challenge.

In another aspect of the invention there is provided a method of screening a library of nucleotide sequences, said method including the steps of: providing a source of nucleotide sequences; providing an expression vector including a marker, preferably a polyHis tag; WO 00/31115 PCT/AU99/01035 17 creating a library by introducing a nucleotide sequence into the expression vector; identifying an expression protein expressed by the expression vector by determining the presence of a fusion protein comprising a marker co-expressed with the expression protein translated by the nucleotide sequence; and determining the nucleotide sequence of the expression protein gene.

In another aspect, there is provided a method of screening to identify expression clones in a library wherein said method includes: providing a source of nucleotide sequences; providing an expression vector including a marker, preferably a polyHis tag; introducing a nucleotide sequence into the expression vector; transforming the expression vector into a host to create an expression library; and locating an expression clone by detecting the expression of a fusion protein, comprising a marker, preferably a polyHis tag co-expressed with an expression protein.

Identification of the expression proteins having a polyHis tag transcribed by the expression clones further identifies those clones (host cells) which express an open reading frame (ORF).

Accordingly, the method identifies ORF's in a mixture of nucleotide sequences. This effectively speeds up the process of identifying useful nucleotide sequences encoding proteins which are potentially useful as antigens. It quickly identifies a clone which can be immediately expanded to provide sufficient sample of nucleotide sequence or plasmid DNA or recombinant protein for any use such as in genetic vaccines or for developing antibodies against a recombinant protein.

In a further aspect of the present invention there is provided a method of identifying a therapeutic antigenic gene sequence encoding a therapeutic antigenic protein of a disease, from a sample of nucleotide sequences, said method comprising the steps of: providing an expression vector including a marker; WO 00/31115 PCT/AU99/01035 18 introducing a nucleotide sequence from the sample into the expression vector; identifying an antigenic expression protein expressed by the expression vector by determining the presence of a fusion protein comprising a marker, coexpressed with the expression protein translated by the nucleotide sequence; inoculating an animal with the expression vector which expresses the antigenic expression protein; challenging the animal with the disease; identifying an expression vector and expression protein that provides a therapeutic effect induced by vaccination; and determining the gene sequence of the antigenic expression protein encoded by the expression vector.

It is an advantage of the invention that it provides an efficient means of screening a library of nucleotide sequences and using identified clones for DNA vaccination trials to immediately assess the vaccine potential of clones. This aspect of easily using the identified clones to do vaccine trials far improves methods presently available.

In a further preferred embodiment of the invention, there is provided a method of screening a library of a nucleotide sequence for a nucleotide sequence encoding polypeptides of Mycoplasma, preferably Mycoplasma hyopneumoniae, said method comprising the steps of: providing a source of nucleotide sequences from a sample of Mycoplasma; providing an expression vector including a marker, preferably a polyl-lis tag; introducing a nucleotide sequence into the expression vector; identifying a Mycoplasma expression protein expressed by the expression vector by determining the presence of a fusion protein comprising a marker, preferably a polyHis tag, co-expressed with the expression protein translated by a nucleotide sequence; and determining the nucleotide sequence of the expression protein gene of Mycoplasma, preferably Mycoplasma hyopneumoniae.

WO 00/31115 PCT/AU99/01035 19 The source of nucleotide sequence from a sample of Mycoplasma may be of genomic, mitochondrial, recombinant or mRNA origin. Where the source is genomic, genomic DNA may be isolated from Mycoplasma by any means known to the skilled addressee.

The Mycoplasma is preferably Mycoplasma hyopneumoniae. The methods of identification of the polyHis tag-Mycoplasma fusion protein are as described above.

Preferably, the nucleotide sequence encodes an antigenic protein.

Accordingly, there is a further step of determining whether the protein elicits an immune response. Preferably, the antigenic protein is a Mycoplasma antigenic protein. More preferably a Mycoplasma hyopneumoniae antigenic protein.

The term "immune response" means a selective response mounted by the immune system in which specific antibodies and/or cytotoxic cells are produced against invading of foreign components such as microorganisms, parasites, transplanted tissue, proteins or antigens in general.

The term "immune system" includes any cells and tissues which enable the mounting of a specific protective response to invading of foreign components such as microorganisms, parasites, transplanted tissue, proteins or antigens in general aimed at protecting the body from infection and in some cases setting up a long lasting specific immunity to re-infection.

In yet another aspect of the present invention, there is provided a nucleotide sequence mutant, analogue, derivative or functionally active fragments thereof encoding a polypeptide of Mycoplasma, preferably Mycoplasma hyopneumoniae, said nucleotide sequence identified by the methods described above. Preferably the nucleotide sequence comprises a sequence according to any one of Figures 1 to 42, and mutants, analogues derivatives or functionally active fragments thereof. More specifically it comprises SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:41, SEQ ID NO:43, SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53, SEQ ID NO:55, SEQ ID NO:57, SEQ ID WO 00/31115 PCT/AU99/01035 NO:59, SEQ ID NO:61, SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:67, SEQ ID NO:69, SEQ ID NO:71, SEQ ID NO:73, SEQ ID NO:75, SEQ ID NO:77, SEQ ID NO:79, SEQ ID NO:81 or SEQ ID NO:83 or a mutant, analogue, derivatives or functionally active fragments thereof.

In another embodiment there is provided a recombinant vector including a nucleotide sequence described above, mutant analogue derivative or functionally active fragment thereof.

according to any one of Figures 1 functionally active fragment thereof.

NO:1, SEQ ID NO:3, SEQ ID NO:5, NO:11, SEQ ID NO:13, SEQ ID NO:15, NO:21, SEQ ID NO:23, SEQ ID NO:25, NO:31, SEQ ID NO:33, SEQ ID NO:35, NO:41, SEQ ID NO:43, SEQ ID NO:45, NO:51, SEQ ID NO:53, SEQ ID NO:55, NO:61, SEQ ID NO:63, SEQ ID NO:65, NO:71, SEQ ID NO:73, SEQ ID NO:75, Preferably the nucleotide sequence to 42, mutant analogue derivative More specifically it comprises SEQ SEQ ID NO:7, SEQ ID NO:9, SEQ SEQ ID NO:17, SEQ ID NO:19, SEQ SEQ ID NO:27, SEQ ID NO:29, SEQ SEQ ID NO:37, SEQ ID NO:39, SEQ SEQ ID NO:47, SEQ ID NO:49, SEQ SEQ ID NO:57, SEQ ID NO:59, SEQ SEQ ID NO:67, SEQ ID NO:69, SEQ SEQ ID NO:77, SEQ ID NO:79, SEQ NO:81 or SEQ ID NO:83 or a mutant, analogue, derivatives or functionally active fragments thereof.

The term "mutant, analogue, or derivative" refers to those sequences that may contain deletions, additions or substitutions of their sequences.

Where these terms relate to a protein, the changes do not substantially change the activity of the protein. Where it relates to a changed nucleic acid molecule, the change does not result in a change in the reading frame of a protein coding region and preferably encodes a protein having no change, only a minor reduction or an increase in biological function.

The term "fragments" relates to a portion of an amino acid or nucleic acid sequence that is less than full length but is capable of hybridizing to a full length sequence and being recognised as being part of a nucleic acid sequence capable of encoding an amino acid sequence which is a portion of the full length sequence.

Due to the degeneracy of the genetic code, other nucleic acid sequences which encode the same or functionally equivalent amino acid WO 00/31115 PCT/AU99/01 035 21 sequence are included within the scope of the current invention. Such alterations of the nucleotide sequence may include substitutions of different nucleotides resulting in the same or a functionally equivalent gene product.

Also included within the scope of this invention are nucleic acid sequences having deletions and/or additions and which result in a functionally equivalent gene product. In addition the gene product may include deletions, additions or substitutions of amino acid residues within the sequence which result in changes that still produce a functionally active product.

In a further embodiment, there is provided a host cell including a recombinant vector described above. The host cell may be any cell competent to receive the recombinant vector. Preferably the host cell is prokaryotic, more preferably it is E. coli.

In another aspect of the invention there is provided an amino acid sequence which encodes a polypeptide identified by the methods described above. The polypeptide may be a product of the expression vector or is an expression protein identified as a protein co-expressed with a marker preferably a polyHis marker. Preferably, the amino acid sequence is translated from a nucleotide sequence identified by the methods described above.

In a preferred aspect, the amino acid sequence encodes a polypeptide of Mycoplasma, preferably Mycoplasma hyopneumoniae. More preferably, the amino acid sequence comprises a sequence according to any one of Figures 1 to 42 or a mutant, analogue, derivative, or functionally active fragment thereof.

More specifically it comprises SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO:40, SEQ ID NO:42, SEQ ID NO:44, SEQ ID NO:46, SEQ ID NO:48, SEQ ID NO:50, SEQ ID NO:52, SEQ ID NO:54, SEQ ID NO:56, SEQ ID NO:58, SEQ ID NO:60, SEQ ID NO 62, SEQ ID NO:64, SEQ ID NO:66, SEQ ID NO:68, SEQ ID NO:70, SEQ ID NO:72, SEQ ID NO 74, SEQ ID NO: 76, SEQ ID NO:78, SEQ ID NO:80, SEQ ID NO:82 or SEQ ID NO:84 or a mutant, analogue, derivatives or functionally active fragments thereof.

WO 00/31115 PCT/AU99/01035 22 In an even further aspect of the present invention, there is provided a polypeptide encoded by the amino acid sequence mutant, analogue, derivative or functionally active fragment thereof as described above. Preferably the sequence encodes a polypeptide of Mycoplasma, preferably Mycoplasma hyopneumoniae, said amino acid sequence being identified by the methods described above. Preferably the polypeptide is encoded by an amino acid sequence according to any one of Figures 1 to 42, mutant, analogues, derivatives or functionally active fragment thereof. More specifically it comprises SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID SEQ ID NO:32, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:38, SEQ ID SEQ ID NO:42, SEQ ID NO:44, SEQ ID NO:46, SEQ ID NO:48, SEQ ID SEQ ID NO:52, SEQ ID NO:54, SEQ ID NO:56, SEQ ID NO:58, SEQ ID SEQ ID NO 62, SEQ ID NO:64, SEQ ID NO:66, SEQ ID NO:68, SEQ ID SEQ ID NO:72, SEQ ID NO 74, SEQ ID NO: 76, SEQ ID NO:78, SEQ ID SEQ ID NO:82 or SEQ ID NO:84 or a mutant, analogue, derivatives or functionally active fragments thereof.

The polypeptide may be isolated and purified by any methods known to the skilled addressee. Preferably, the methods of isolation utilise the polyHis tag which was used to identify expression protein or expression clones from a library of nucleotide sequences.

In a further embodiment, there is provided a composition comprising the polypeptide or nucleotide sequence as described above. The composition may be a pharmaceutical composition comprising at least one polypeptide or nucleotide sequence as described above and a pharmaceutically acceptable carrier. The composition may also be a vaccine composition comprising at least one polypeptide or nucleotide sequence described above. The vaccine composition may be capable of eliciting an immune response wherein the polypeptide is an antigenic protein or a nucleotide sequence preferably of Mycoplasma, more preferably of Mycoplasma hyopneumoniae.

WO 00/31115 PCT/AU99/01035 23 In another aspect of the present invention, there is provided an antibody against a polypeptide described or identified according to the methods described above. Preferably the antibody is a monoclonal antibody.

Various procedures are known in the art which may be used for the production of antibodies to epitopes of the polypeptides described above.

Various host animals may be used in the production of these antibodies following immunisation with the polypeptides including but not restricted to rabbits, mice, goats etc. Adjuvants may be used to increase the immunological response, depending on the host species, and may include but are not restricted to Freunds (complete and incomplete). 'Monoclonal antibodies may be prepared by using techniques which enable the continuous production of antibody molecules by cell lines in vitro. These may include, but are not limited to, the hybridoma technique (Kohler and Milstein, Nature 256, 495, (1975)).

Antibodies to the polypeptides may find use in the detection of antigenic polypeptides in various tissues, body fluids and cell lines, for example in screening assays for the antigen, and in the affinity purification of polypeptides, preferably antigenic polypeptides.

The vaccine composition includes purified plasmid DNA including nucleotide sequences described above.

In yet another aspect of the present invention, there is provided a method of treating an infection, said method including administering an effective amount of an antigenic polypeptide encoded by an amino acid sequence said polypeptide being identified by the methods described above.

Preferably the infection is a Mycoplasma infection. More preferably, the infection is a Mycoplasma hyopneumoniae infection.

Also it is preferred that the polypeptide is encoded by an amino acid sequence encoded by a nucleotide sequence according to any one of Figures 1 to 42, mutant, analogue, derivative or functionally active fragment thereof.

More specifically the sequence comprises SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33, SEQ ID SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:41, SEQ ID NO:43, SEQ ID WO 00/31115 PCT/AU99/01035 24 SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53, SEQ ID SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:61, SEQ ID NO:63, SEQ ID SEQ ID NO:67, SEQ ID NO:69, SEQ ID NO:71, SEQ ID NO:73, SEQ ID SEQ ID NO:77, SEQ ID NO:79, SEQ ID NO:81 or SEQ ID NO:83 or a mutant, analogue, derivatives or functionally active fragments thereof.

It is also preferred that the polypeptide has an amino acid sequence according to any one of Figures 1 to 42, mutant, analogue, derivative or functionally active fragment thereof. More specifically it comprises SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO:40, SEQ ID NO:42, SEQ ID NO:44, SEQ ID NO:46, SEQ ID NO:48, SEQ ID NO:50, SEQ ID NO:52, SEQ ID NO:54, SEQ ID NO:56, SEQ ID NO:58, SEQ ID NO:60, SEQ ID NO 62, SEQ ID NO:64, SEQ ID NO:66, SEQ ID NO:68, SEQ ID NO:70, SEQ ID NO:72, SEQ ID NO 74, SEQ ID NO: 76, SEQ ID NO:78, SEQ ID NO:80, SEQ ID NO:82 or SEQ ID NO:84 or a mutant, analogue, derivatives or functionally active fragments thereof.

In yet another aspect of the present invention, there is provided a method of treating an infection, said method including administering to an animal in need, an effective amount of a DNA molecule including a nucleotide sequence identified by the methods described above. Preferably the infection is a Mycoplasma infection. More preferably, the infection is a Mycoplasma hyopneumoniae infection.

Preferably, the nucleotide sequence is according to any one of Figures 1 to 42, mutant, analogue, derivative or functionally active fragment thereof or more specifically, SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:41, SEQ ID NO:43, SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53, SEQ ID NO:55, SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:61, SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:67, WO 00/31115 PCT/AU99/01 035 SEQ ID NO:69, SEQ ID NO:71, SEQ ID NO:73, SEQ ID NO:75, SEQ ID NO:77, SEQ ID NO:79, SEQ ID NO:81 or SEQ ID NO:83 or a mutant, analogue, derivatives or functionally active fragments thereof.

The DNA may be administered by injection which allows for cells to take up DNA or it may be applied topically. Preferably, the DNA is injected into the skin layer subcutaneously, or intramuscularly.

In another aspect of the present invention, there is provided a method of treating an infection, said method including administering an effective amount of an antibody specific to a polypeptide which has been identified by a method described above.

Preferably the infection is a Mycoplasma infection. More preferably, the infection is a Mycoplasma hyopneumoniae infection.

Preferably the polypeptide is encoded by an amino acid sequence according to any one of Figures to 1 to 42, a mutant, analogue, derivative or functionally active fragment thereof or more specifically SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO:40, SEQ ID NO:42, SEQ ID NO:44, SEQ ID NO:46, SEQ ID NO:48, SEQ ID NO:50, SEQ ID NO:52, SEQ ID NO:54, SEQ ID NO:56, SEQ ID NO:58, SEQ ID NO:60, SEQ ID NO 62, SEQ ID NO:64, SEQ ID NO:66, SEQ ID NO:68, SEQ ID NO:70, SEQ ID NO:72, SEQ ID NO 74, SEQ ID NO: 76, SEQ ID NO:78, SEQ ID NO:80, SEQ ID NO:82 or SEQ ID NO:84 or a mutant, analogue, derivatives or functionally active fragments thereof.

The present invention will now be more fully described with reference to the accompanying Examples and Figures. It should be understood, however, that the description following is illustrative only and should not be taken in any way as a restriction on the generality of the invention described.

WO 00/31115 PCT/AU99/01035 26

EXAMPLES

EXAMPLE 1: Construction of genomic libraries in pCI31 and pCI32.

Genomic DNA isolated from Mycoplasma hyopneumoniae was broken down into suitably sized fragments by sonication or partial restriction enzyme digestion. The DNA was then run on an agarose gel and size selected fractions cut from the gel and recovered. Vector DNA (pCI30, pCI31, and pCI32) was digested with an appropriate restriction enzyme (Sma I or BamHI) treated with shrimp alkaline phosphatase to remove the terminal phosphates, then run on an agarose gel. The linearized band was cut out and the DNA recovered.

Genomic DNA fragments and cut vector DNA were mixed at molar ratios of between 1:1 and 5:1 and ligated using standard ligation conditions. The ligation mixtures were used to transform competent E. coli cells and thus produce a number of distinct genomic libraries.

EXAMPLE 2: Screening of the genomic libraries.

The genomic libraries were plated onto Luria-Bertoni (LB) broth agar containing ampicillin (100 pg/ml) and grown overnight at 370C. Filter lift replicas of the plates were made by placing dry; sterile nitrocellulose membranes on top of the colonies then immediately removing it and placing it colony side up onto an LB plate containing ampicillin and IPTG (1mM) and incubating for 4 hours at 37 0 C. The membranes were then processed by standard methods to lyse the bacteria, neutralise the membranes and bind protein to the membranes. The dry membranes were then probed either with anti-polyHis antibodies or with conjugated nickel resin and the polyHis signals developed by addition of chromogenic substrate. The developed membranes were keyed back to the original plates and the positive colonies picked in to 96 well, deep well plates containing rich growth media supplemented with ampicillin. The clones were then grown at 370C. Replicas of these plates were taken and held at -70°C for long term storage.

EXAMPLE 3: Sequence analysis of selected clones.

A collection of polyHis positive clones from each library was selected.

Plasmid DNA was prepared and DNA sequence information from the inserted fragment was generated by automated dideoxy sequencing using a primer WO 00/31115 PCT/AU99/01035 27 homologous to a region just upstream of the site of insertion into the vector.

The sequence data generated was compared with the publicly available GenBank database. Homologies between our positive sequences and database entries were noted and in many instances this information gave some indication of the possible function of the gene fragments which had been cloned. On the basis of this homology information a subgroup of the positive clones was selected which were likely to represent genes which encoded proteins that may be easily accessible to the host immune system during Mycoplasma hyopneumoniae infection or that for other reasons were regarded as likely to offer some level of protection. These selected positive clones encode candidate protective antigens.

Some of the clones chosen for further study represent different sections of the same gene.

EXAMPLE 4: Screening of candidate protective antigens A series of animal trials were conducted to investigate the therapeutic application of the identified genes. All the trials followed a similar protocol. Pigs from a M. hyopneumoniae free herd were vaccinated 3 times, at 2 week intervals, starting from 6 weeks of age. Each vaccination was with 500 .g of plasmid DNA, divided evenly between 2 shots, 1 shot into the tissue at the base of the back of each ear, delivered using a Biojector 2000 needless injection device with a No.2 syringe. 1-2 weeks after the third vaccination the pigs were challenged by intratracheal instillation of a 5 ml dose of 0.45 gM filtered lung homogenate derived from a pig with an active M. hyopneumoniae infection. 3-4 weeks after challenge the pigs were euthanased and the lungs removed for scoring of pneumonic lesions. The pneumonic lesions were scored by counting the number of lung lobes that were affected and by estimating the percentage of each lung lobe that was affected by disease. This scoring system gives two measures of disease status, a lobe score and a lesion score. For each trial the average results across each group are presented. The trial results of those groups of clones which had a positive therapeutic effect are presented. Other groups of clones did not give a positive therapeutic effect and these results are not presented.

WO 00/31115 PCT/AU99/01035 Trial 1 Vaccinated with a group of clones that included clones pAD612-789.

Treatment Unvaccinated, unchallenged Unvaccinated, challenged Vaccinated, challenged Lobe score Lesion score 0 0 4.4 120 3.25 72.5 The vaccine containing clones pAD612-789 had a therapeutic effect. The lobe score in the vaccinated pigs was 26% lower than in the controls and the lesion score was 40% lower.

Trial 2 Two different groups of clones tested as vaccines.

Group 1: pAD's 662, 774, 784, 922, and 964 Group 2: pAD's 612, 633, 639, 640, 641, 653, 657, 659, 681, 700, 711, 721, 727, 742, 760, 789, 908, 910, 911, 913, 920, 923, 925, 950, 951, 966, 967, 977, 981, 983, 984, 994, 1001, 1005, 1013, 1016, 1020, 1027, 1033, 1037, 1038, 1040, and 1049.

Treatment Unvaccinated, unchallenged Vector vaccinated, challenged Group 1 vaccinated, challenged Group 2 vaccinated, challenged Lobe score Lesion score 0 0 3.4 67 1.6 38 2.2 51 The overall severity of disease induced in trial 2 was less than in trial 1 indicating a milder challenge. Both groups of clones had a therapeutic effect.

WO 00/31115 PCT/AU99/01035 29 The Group 1 vaccine reduced the lobe score by 53% and the lesion score by 43%. The Group 2 vaccine reduced the lobe score by 35% and the lesion score by 24%, compared with the group vaccinated with the empty vector plasmid

DNA.

Trial 3 Group 1: Same clones as Group 1 in trial 2.

The DNA was delivered in the standard way as detailed for the biojector or by genegun. For genegun delivery 1.6 micron gold particles were used with each pig receiving 4 shots, each of 1.25 p.g DNA, at each of the 3 vaccination time points. Vaccination was behind each ear and the inside of each hind leg. The delivery pressure was 500 psi.

Treatment Lobe score Lesion score Unvaccinated, unchallenged 0 0 Vector vaccinated, challenged 4.2 109 Group 1 biojector vaccinated, challenged 2.5 67 Group 1 genegun vaccinated, challenged 2.2 48.5 This trial again demonstrated that the five clones in Group 1 produce a therapeutic effect when delivered as naked DNA. The biojector delivered vaccine gave a 40% reduction in lobe score and a 39% reduction in lesion score. The same collection of clones delivered by genegun gave a 48% reduction in lobe score and a 56% reduction in lesion score.

Trial 4 Other combinations of the clones used in trial 2 were used in this trial.

Group 3: pAD's 653, 711, 727, 920, 951, and 965 Group 4: pAD's 612, 639, 640, 1005, 1020, and 1038 Group 5: pAD's 742, 760, 923, 925, 994, 984, and 1037 A group of 5 other clones that had not previously been used in trials was also included.

Group 6: pAD's 702, 763, 766, 957, and 996.

Treatment Lobe score Unvaccinated, unchallenged 0 Vector vaccinated, challenged 5.0 Group 3 vaccinated, challenged 2.8 Group 3 genegun vaccinated, challenged 2.6 Group 4 vaccinated, challenged 3.0 Group 5 vaccinated, challenged 3.6 Group 6 vaccinated, challenged 2.75 Lesion score 0 106 54 87 79 74 42.5 All of these groups of clones produced a therapeutic effect in that they reduced 15 the severity of pneumonic lesions. The group 3 clones reduced the lobe score by 44% when delivered by biojector and by 48% when delivered by genegun, and the lesion score by 49% when delivered by biojector and 18% when delivered by genegun. The other three experimental vaccines were delivered only be biojector. The group 4 vaccine reduced the lobe score by 40% and the 20 lesion score by 25%. The group 5 vaccine reduced the lobe score by 28% and the lesion score by 30%. The group 6 vaccine reduced the lobe score by and the lesion score by Trial The following vaccines were delivered by GeneGun in the same manner as detailed for trial 3.

Treatment Unvaccinated, unchallenged Vector vaccinated, challenged pAD662 pAD784 pAD922 pAD964 Lobe score 0 4.6 1.0 3.4 4.6 1.2 Lesion score 0 163 23 134 124 17 The four clones tested for therapeutic efficacy in this trial are from the Group 1 clones previously tested in Trials 2 and 3 as part of a larger group of 5 clones.

Clone pAD662 gave a 78% reduction in lobe score and an 86% reduction in lesion score. Clone pAD964 gave a 74% reduction in lobe score and a reduction in lesion score. The other clones had little therapeutic effect, giving results similar to that found with the vector control group.

This trial and the previous trials demonstrate that the methods outlined can be used to identify cloned genes that have therapeutic effects as individually delivered genetic vaccines or as components of larger groups of clones.

Trial 6 The recombinant proteins encoded by clones pAD662 and pAD964 were isolated and purified using the polyHis sequences for affinity chromatography.

The recombinant protein solutions were combined and formulated in Fortasol adjuvant from Intervet International BV. Pigs were vaccinated 3 times at twoweek intervals then challenged and lung lesions scored in a similar manner as for the previous trials. Two hundred micrograms of each antigen was present in 20 each dose of vaccine.

Treatment Lobe score Lesion score Unvaccinated, unchallenged 0 0 Unvaccinated, challenged 6.4 158 pAD662 and pAD964 proteins 2.2 A therapeutic effect produced by the recombinant proteins was apparent. The test vaccine group gave a 66% reduction in lobe score and a 75% reduction in lesion score.

This trial demonstrates that antigens identified by the methods outlined, which initially involve screening clones for protein expression followed by genetic vaccination, can also be effective when utilised as recombinant protein based vaccines.

31 This series of trials demonstrates that compared with unvaccinated or empty vector vaccinated pigs the collection of cloned genes detailed in this patent can produce a therapeutic effect by lowering the severity of lung lesions in the groups vaccinated with various combinations of the recombinant plasmids. On the basis of this demonstration of therapeutic effect a collection of 42 genes are claimed in the present invention.

EXAMPLE 5: Screening of genomic libraries to recover the complete genes.

The clones listed in example 4 contain only partial gene sequences. It is desirable to identify the nucleotide sequence of the complete genes, as these are likely to encode further epitopes that will be useful in vaccine design. The complete coding regions of the genes represented by the clones in example 4 S 15 were recovered by screening two genomic libraries. The genomic libraries were constructed by ligating fragments created by digestion with the restriction enzymes EcoRI and Bglll into pUC18 vector digested with EcoRI and BamHI respectively. Ligation products were recovered by transformation of the ligation mix into XL2-BlueMRF' competent cells (Strategene) and plating onto Luria- Bertoni agar plates containing 100 pLg/ml ampicillin. Colonies were picked into 96 well plates containing rich growth media, grown overnight and then plasmid DNA was prepared for each. Plasmid DNA from each was denatured and spotted on to nylon membranes. Representative membranes were then probed with digoxigenin labelled PCR products generated from the inserted DNA present in each of the clones listed in example 4. Hybridizing clones were identified and analysed by restriction enzyme digestion, PCR using gene specific primers and by nucleotide sequencing. The resulting gene sequences are detailed in figures 1-42. A number of these sequences represent the complete coding sequence of the genes but for many of the genes only a partial sequence is available.

Finally it is to be understood that various other modifications and/or alterations may be made without departing from the spirit of the present invention as outlined herein.

EDITORIAL NOTE APPLICATION NUMBER 15376/00 This specification does not contain pages 32 to 159.

EDITORIAL NOTE APPLICATION NUMBER 15376/00 The following Sequence Listing pages 1 to 128 are part of the description. The claims pages follow on pages 160 to 164.

WO 00/31115 WO 0031115PCT/AU99/01035 <210> <211> <212> <213> <220> <223> 1 SEQUENCE LISTING 1125

DNA

Artificial Sequence Description of Artificial Sequence:Clone pAD612 <400> 1 atggcaaaac ataaaaaaag tcagctgaag attttatctg caqcaatttg agttttacct aagggcgaaa gaaatctccc aattcttctt gtcttaaata ggttccggqa acaaaagagg gtggccgqat attaatgttc ccagtttcaa ttgaaaaaaq gctggggctc ttttatatcc atcaatgata aagattttta cttatcgaaa aacaaaaaca atgaaacaaa gtggtgggtc ctggttttgg attttcaagc agaaattaac ctqatattac tcccgggatt aaaacattac aagtaactat ttggtqgacc gtcaqcataa taattgatat ttagacttta ctgatccaaa ccgaatttag aaacaaaggc taaaattctg tttagtaaat agctgaggcc gcgaaaacag tagtggcttt ttttggcggc taaaatttat aaaatacgat aacttgctat tggtcgggtt aaaaattgca taaaattccg gggacacaaa acattttact tatcaaccaa tgattattat aaatccaaga tccccgtcaa aaaatgacta gqagttgaaa atttatcatc aaatttaaag tacgataaat tcaggatttg tcacaagaac atcagtttta caatqtgata aattgtcaag cagaacaaaa aagaagtgcc gccggaatcc ggcgggcctt agatccggaa aatactgtcg aaatccggtc attattggcg aaagatgagc aaagaatttc aatcagcatc ctgataaaaa aaatccagga tcggtcatgc attttggcga aaaaatatag tcgagtcaat actgtaaggg gtcggggaat caacttgttc atggaaaaac aggatggaat ctggaqatct atgatattca aagttcccaa caattgttaa atctcaaggt tcaaccaggt aataa actaacagaa tacaaaaaaa aqcctacgaa cgcttttgat tattttttca tcgtccttta tot cggaaaa ttcaqqcgct gcaaactgag agtttgttta tatagttgag gtttatccgc tcatcttgag tgtgaatatg cccaaccggt tgttcttcct tcatttaatt ttttgctcaa 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1125 WO 00/31115 PCT/AU99/01035 2 <210> 2 <211> 368 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence:Deduced protein sequence from clone pAD612 <400> 2 Met Ala Lys Gln Asp Phe Tyr Lys Ile Leu Gly Val Glu Lys Ser Ala 1 5 10 Ser Leu Thr Glu Ile Lys Lys Ala Tyr Arg Asn Leu Val Asn Ile Tyr 25 His Pro Asp Lys Asn Thr Lys Lys Ser Ala Glu Glu Gin Lys Gin Ala 40 Glu Ala Lys Phe Lys Glu Ile Gin Glu Ala Tyr Glu Ile Leu Ser Asp 55 Glu Thr Lys Arg Lys Gln Tyr Asp Lys Phe Gly His Ala Ala Phe Asp 70 75 Gin Gin Phe Gly Gly Gly Ser Ser Gly Phe Ser Gly Phe Asp Phe Gly 90 Asp Ile Phe Ser Ser Phe Thr Ser Gly Phe Gly Phe Gly Gly Ser Gin 100 105 110 WO 00/31115 PCT/AU99/01 035 Giu Gin Lys Tyr Ser Arg Pro 115 Leu Lys Gly Giu Asn Phe Gin Ala Lys 120 125 Ser Ile Leu Gly Lys Giu Ile Ser Gin 140 Ile Tyr 130 Ile Ser Phe Ile Lys 145 Leu Thr Lys Tyr Gln Cys Asp Asn Cys Lys Gly Ser Gly 155 Asn Ser Ser Ser Asp 165 Ile Thr Thr Cys Tyr Asn Cys Gin Gly 170 Arg Gly 175 Met Gin Thr Lys Thr Thr 195 Ile Ala Lys 210 Val Leu Asn Ile Pro Gly Phe Gly Arg 185 Gly Ser Gly Lys Asn 205 Cys Ser Val Cys Val Gin Asn 190 Ile Thr Lys Lys Giu Giu Lys Cys His Gly 215 Lys Thr Ile Val Giu Thr 220 Val Thr Ile Lys Ile Pro Ala Gly Ile Gin Asp Gly Met Phe Ile Arg 225 230 235 240 Val Ala Gly Phe Gly Gly Pro Gly His Lys Gly Gly Pro Ser Giy Asp 245 250 255 Leu His Leu Giu Ile Asn Val Arg Gin His Lys His Phe Thr Arg Ser 260 265 270 WO 00/31115 PCT/AU99/01035 Gly Asn Asp Ile His Val Asn 275 Asn Gin Asn Thr Val Glu Val 290 295 Met Pro Val Ser Ile 280 Ile Asp Ile Ile 285 Leu Lys Lys Val Pro Asn Pro Thr Gly 300 Arg Leu Tyr Asp Tyr 305 Ala Gly Ala Pro Asp Lys Ser Gly Pro Val Asn Val Leu Pro 320 Pro Lys Asn Pro Arg 330 Ile Ile Gly Asp Leu Lys 335 Val His Leu Ile 340 Glu Leu Asn Gin 355 Phe Tyr Ile Pro Glu 345 Val Phe Ala Gin Ile 360 Phe Ser Pro Arg Gin Lys Asp 350 Asn Asp Lys Thr Lys Ala Lys 365 <210> 3 <211> 819 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence:Clone pAD633 <400> 3 WO 00/31115 WO 0031115PCT/AU99/O1 035 atqataaaag aaaaatatta tcaggaaaat attgaagttg ggtataattt gctttcagtt ttagctgcaa ggccaaaagc tttgatgaaa gatgatctaa gcaattttag ccaaaattaa atctataaaa ctagagcaac tttccgatgt atgttgtttt caacgttttc atggagtttt ttcaaaaccc tggaaaacat aagtgcaaat aaaaagtagc taacttcaat gaaaaqataa ctgatgaaat ttttaaatga tttccagcgc tatgaaaatt ttgctttagt tqaaaaagqt taagattctt actaaataag agataatcaa taatgaggat ggagtcatat aattgcatca gcttgatccc aacaaaaact tattqttttt taaaaatatc acttaaatta aaagcaaaaa tatacaaaca aaatattatg tcaggaattt gaaaatttaa tttgttgggg ccaaaaaaaa ttagaccgtg gttttagcac agaggtaaat ttaatttcaa gcaaatgggg atcgaaaaag gttagtccaa acatcgtaa acatggacca caattctagg ttaaacctca cgaaaattag caacggttga tgagtcaaat agccacaatt taaatcctga atgatgttgt tcacccacaa gaattatcgc cgaaaattga cttatgacga gcttgtgctg gcataatggt aaaaggtagt gaaaaaaatt agatgacatc aatcgcaaat tttatctggg gattataatt taaaattctt tatgaatgaa tcagggggat ctccccattt aaatgaattg 210> <211> <212> <213> <220> <223> 4 272

PRT

Artificial Sequence Description of Artificial Sequence: Deduced protein sequence from clone pAD633 <400> 4 Met Ile Lys Val Ser Asp Val Cys Phe Ser Tyr Thr Asn Asn Met Asp 1 5 10 Gin Leu Val Leu Lys Asn Ile Asn Val Val Phe Glu Lys Gly Lys Tyr 25 WO 00/31115 WO 0031115PCT/AU99/01035 Tyr Ala Ile Leu Giy His Asn Ile Leu Ser Gly Ile Phe Lys Gly Val Leu Leu Asn Lys Glu 70 Giy Ser Gly Lys Ser 40 Thr Phe Ser Lys Ile Giu Val Asp Pro Gin Lys Gly Asn Leu Thr Lys 7 Ile Arg Lys Lys Gly Ile Ile Phe Gin Asn Pro Asp Asn Gin Phe 90 Val Gly Aia Thr Vai Giu Asp Asp Ile Ala Phe Ser Leu Glu Asn Ile Asn Giu Asp Pro Lys 100 105 110 Lys Met Ser Gin Ile Ile Ala Asn Leu Ala Ala Lys 115 120 Val Gin Met Giu 125 Gly Gin Lys Gin Ser Tyr Leu Asp Arg Giu Pro Gin Phe Leu Ser 130 135 Lys Val Ala Ile Ala Ser Val Leu Ala Leu Asn 145 150 155 Phe Asp Giu Ile Thr Ser Met Leu Asp Pro Arg 165 170 Pro Glu Ile Ile Gly Lys Tyr Asp Val 175 Val Lys Ile Leu Asp Asp Leu Arq Lys Asp Lys Thr Lys Thr Leu Ile 180 185 190 WO 00/31115 WO 0031115PCT/AU99/01 035 Ser Ile Thr His Asn Me-t 195 Asn Glu Ala Ile Leu Ala 200 Ile Ile Ala Gin Gly Asp 215 220 Val Phe 210 Ala Asn Gly Gly Asp Giu Ile Ile 205 Pro Lys Leu Ile Asp Ser Pro Phe 240 Asn Asp Lys Asn Ile Tyr Lys Ile Ser 245 Glu Asn Giu Leu Leu 260 Ile Ile Giu Lys Ala 230 Ser Ala Leu Lys Leu 250 Giu Gin Leu Trp Lys 265 Lys Ile 235 Vai Ser Pro Thr Tyr Asp 255 Leu Lys Gin Lys Thr Ser 270 <210> <211> <212> <213> <220> <223> 609

DNA

Artificial Sequence Description of Artificial Sequence:Cione pAD639 <400> atgtttagtc aaactattta taaatgaaaa agcgaaaatc tcgaaagaqt ggtcgtccct gaaaaaggac tttcttgcga atataacggt ctttttaqca taagaactgg caaaagtttg 120 WO 00/31115 WO 0031115PCT/AU99/01 035 tacgggctgc cttatcttta gatttttqat tttaaaataa aaccggtttt tcaaacttat gatttaaata gaaactttag ttagagatc tacaaaattt cctcttccgc tgcataaaaa tggatggtag tacaagaact tttttgtcag attcaactat ctctaaaaag agataatgat atcatccgct taatttttta atgagaaaat aaaaagjcaaa cctgagtcta ttatttaaat tcttggaaat tattttattg ttagtaaqag gtcggcctta acctatcttg aaacactata actaatcata gaattatgcc tatgattttg cttacctaag caaaqtctta tcgtctttag ttaaatcagg aattagaatg tcagaagtct ttgatgatct acgatcaaga atcaagtgtg ttgcgaaaac tgccgqaatt ggatqgtttt ttttttgctt agcccatcca tagtcaaaaa aaaggaactt 180 240 300 360 420 480 540 600 609 <210> 6 <211> 203 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Deduced protein sequence from clone pAD639 <400> 6 Met Phe Ser Gin Thr Ile Tyr Lys Trp Lys Ser Glu Asn Leu Giu Arq 1 5 10 Val Val Val Pro Glu Lys Gly Leu Ser Cys Glu Tyr Asn Gly Leu Phe 25 Ser Ile Arg Thr Gly Lys Ser Leu Tyr Gly Leu Leu Gin Asn Leu Asp 40 Asn Asp Tyr Phe Ile Ala Tyr Leu Arg Ser Ser Val Leu Ile Phe Thr 55 WO 00/31115 WO 0031115PCT/AU99/O1 035 Ser Ser Ala Ser Ser Ala Leu Val Arg Ala Lys Ser Tyr Cys Glu Asn 70 75 Lys Asn Asn Phe Leu Val Gly Leu Ile Vai Phe Asp Phe Trp Leu Ser Ala Gly Leu Val Lys 115 Phe Lys Ile Ser Gly Asp Gly Met Asp Gly Arg Trp Glu 105 Phe Asn Arq Phe Leu Gin 120 125 Giu Cys Phe Leu Leu Ser 140 Asn Thr Tyr 110 Glu Leu Lys Ser Lys 130 Lys His Tyr Lys Leu 135 Asn Leu Phe Phe 145 Val Ser Leu Ser Thr Asn His Ile Arg Ser Leu Ala His Pro 155 160 Giu Leu Cys Leu Asp Asp 175 Asp Leu Asn Asn Leu Ser Gin Lys 180 Phe Asp Asp Gin 195 Thr Ile Tyr Leu Giu Thr Leu Ala Leu 185 Lys Ser Leu Giy Asn Tyr Asp 190 Giu Lys Glu Leu Leu Giu Ile 200 <210> 7 WO 00/31115 WO 0031115PCT/AU99/01 035 <211> 471 '<212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence:Clone pAD640 <400> 7 gaqgtaaqca atgcaagaat attaattttg aaagatgggc aagccggtct gcaatcggcg tttattccca atccagttta cgattcccgg cagcccggat attttgaaaa catcagcagg cacataatat qactaaaaga aggcgaacga ttcccgttga acgaggcgat tgccctttct cactttaatt gataactttt tgcaatgaca aaagacgatg gaaaaattta tacttatcaa attaaactaa tatgttcagt catattcatg gcaacaqcaa ggqgagataa ggggcctatt gtcgatattc ccaatttatg caggttcgct caaaggccaa tacccgaag g taatttcagc ccttgcgcgg taaatgggat cgcaggaagt attttatttt taaggatqta ggattttggg agcaattcca cctctcgcaa gaaggtttta taaaattatt aaaaqacgtt t <210> 8 <211> 157 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Deduced protein sequence from clone pAD64O <400> 8 Glu Val Ser Thr Ile Pro Gly Arg Gly Asp Ile Lys Leu Thr Gly Ser 1 5 10 Leu Lys Asp Val Met Gln Glu Ser Ala Arq Ile Ala Leu Ser Tyr Val WO 00131115 WO 0031115PCT/AU99/O1 035 Gin Ser Lys Ala Lys Asp Phe Gly Ile Asn Phe Asp Phe Giu Asn Thr 40 Leu Ile His Ile His Val Pro Glu Gly Ala Ile Pro Lys Asp Giy Pro 55 Ser Ala Giy Ile Thr Phe Ala Thr Ala Ile Ile Ser Ala Leu Ser Gin 70 75 Lys Pro Val Ser His Asn Ile Ala Met Thr Gly Glu Ile Thr Leu Arg 90 Gly Lys Val Tyr Leu Asn 115 Ala Ile Gly Gly Leu Lys Giu Lys Thr 105 Met Gly Aia 110 Asn Giu Lys Gly Ile Lys Ile Ile 120 Phe Ile Pro Lys Asn Leu 130 Pro Val 145 Val Asp Ile Pro Gin 135 Asp Thr Tyr Gin Pro 150 Giu Val Lys Asp Vai 140 Ile Gin Phe Ile Ile Tyr Asp Phe Ile Phe 155 <210> 9 <211> 1128 <212> DNA WO 00/31115 WO 0031115PCT/AU99/01 035 12 <213> Artificial Sequence <220> <223> Description of Artificial Sequence:Clones pAD64l and pAD1O33 <400> 9 aaaaaacttt tagatacttt tcgatctcac atcatttttg cttgttgaaa gaaattaaga aagccaataa atcgagcagc agaaatttat ttagtttttt gatatgcctt attaaggaca ctaaatgtta attttttctt attaaacaat ggaaatcaac ctcgatgaac caccaattaa atcggaatcg aaaagcaatt atgaaccgac.

aacttaaaaa aaacttgtga atgaatttaa gctatcccga caaatqataa atggccttgt atttaaatgg gtcttgatca atttaccaat tccttccttt taaagatcaa aaaaagtttc caactcgcg aacaagaaaa ctgatcgggt aggggctgaa tatcgaaatc ttcggttcta acaaggaatt atttatcact aaacgaagat aaaaacgccq agtttctaat aagttcaggt tcatattgaa tggaatttat taattttaat tgtctcaaaa aacaacttca acttgcaaaa agtcgatgtt taaaacaatt aattacaatg tttagtccaa gccaaagctt accgaaaaag gcaatcqtct qtgattcgaa gagattattt gttagaagta atcagttttg cgaactgaat ctaaatggtc tatctttctg atcacaattt gcaaaaataa caagatacgc gggcttgcaa ggcgcaagaa atgataattt tatgaaggca aagcacttgt tatcccaaaa atacccaaaa gaattaccca acggaatgta ctttaatgqt aaaaaccatc aaatcaaacc tagctagaac aagaatttcg aaaatagqaa cttctcttgg ctaaaactac cattaacttc cccaaccgca aggaaattta cttcggatat gaattacaag ttcttcgctt accggaaatt actttatctg tagaatggaa tattgaaagt cggttttgat qt ttt tagtt agqtgaaatt tttaattggc cccaaaaaat acaaattggt ctcaaatcag aaatcattat tttatcaggt agttttcatc taatttaatt gcaagaggtt tgaattagtt 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1128 ggcccgcaaa ttaccgatca aaatataatg aaatattcac ttaattta <210> <211> 376 <212> PRT <213> Artificial Sequence WO OW31115 WO 0031115PCTAU99/01 035 13 <220> <223> Description of Artificial sequence: Deduced protein sequence from clones pAD64i and pADlO33 <400> Lys Lys Leu Leu Asp Thr Leu Gly Ala Glu Phe Ser Pro Lys Ala Leu 1 5 10 Val Ser Ser Leu Ser Ile Ser Gin Lys Gin Phe Ile Glu Ile Ala Lys 25 Ala Leu Ser Gin Lys Pro Gu Ile Ile Ile Phe Asp Glu Pro Thr Ser 40 Val Leu Thr Giu Lys Asp Thr Gin Lys Leu Tyr Leu Leu Val Giu Lys 55 Leu Lys Lys Gin Gly Ile Ala Ile Val Trp Ile Thr His Arg Met Glu 70 75 Glu Ile Lys Lys Thr Cys Giu Phe Ile Thr Val Ile Arg Asn Gly Met 90 Tyr Ile Giu Ser Lys Pro Ile Asn Giu Phe Lys Asn Glu Asp Giu Ile 100 105 110 Ile Ser Leu Met Val Gly Phe Asp Ile Giu Gin Arg Tyr Pro Giu Lys 115 120 125 WO 00/31115 WO 0031115PCT/AU99/O1 035 Thr Pro Val Arg Ser 130 Asn 145 Asp Lys Val Ser Lys Lys Pro Ser Phe Leu Val Arg Asn Leu Ser 135 140 Asn Ile Ser Phe Giu Ile Lys Pro Gly Giu Ile 150 155 160 Leu Val Ser Ser Gly Arg Thr Giu Leu Ala Arg 170 175 Leu Val Phe Tyr Thr Leu Ile Gly 180 Asp Met Pro Tyr Leu Asn Gly His Ile Giu Leu Asn 185 190 Gly Gin Giu 195 Phe Arg Pro Lys Asn Ile Lys Asp Ser 200 Arg Lys Gin Ile Giy 220 Leu Asp His Gly 205 Leu Asn Val Asn Ile Tyr 210 Leu Pro 225 Tyr Leu Ser Giu Ile Asn Phe Ile Thr Ile Ser Ser Leu 235 Gly Ser Asn Gin 240 Ile Phe Ser Phe Leu 245 Pro Phe Val Ser Lys Ala Lys Ile Thr 250 Lys Thr 255 Thr Asn His Thr Pro Leu 275 Tyr 260 Ile Lys Gin Leu Lys Ile Lys Thr Thr 265 Gly Asn Gin Gin Lys 285 Ser Gin Asp 270 Val Ser Leu Thr Ser Leu Ser Gly 280 Ala Lys Gly Leu Ala Thr Gin Pro Gin Val Phe Ile Leu Asp Giu Pro WO 00/31115 WO 0031115PCT/AU99/01 035 300 Thr Arg Gly Val Asp Val Gly Ala Arg Lys Giu 305 310 315 Ile Tyr Asn Leu His Gin Leu Lys Gin Glu Asn Lys Thr 325 Met Gin Giu Val Ile Gly Ile Ala Asp 340 345 Gly Arg Ile Thr Ser Giu Leu Val Gly 355 360 Ile Met Lys Tyr Ser Leu Asn Leu 370 375 Ile Met 330 Ile Ile Ser Ser Asp 335 Arg Val Ile Thr Met Tyr Glu 350 Pro Gin Ile Thr Asp Gin Asn 365 <210> <211> <212> <213> <220> <223> 11 2394

DNA

Artificiai Sequence Description of Artificial Sequence:Deduced protein sequence from clone pAD653 <400> 11 acagtcggaa ttaacaaaac cgaaatggat gcaaatacca aaagaatgat gtataatgcc gatattactt attcggttca ttctgaatta ggttttgatt atctccggga taatatggtt 120 WO 00/31115 PCT/AU99/01035 ttttcagcag tcaattttga ccagcccaat attgatgaaq ttttttqgcc aacgcgctqa aaaattgcct ggtctccaqc ctagcaacaa gggactgcca attccgacaa cacgaaaaaa attttaattg aaaggacttt gcccaggcag gatataattt aaagccgaaa gtgggaattt tttqatcaaa catgcggttt actgtgctta gatattttgc gttgatatca aatttaataa attggaattg actatttatt gaatcagaac accattgaca caaattttta caggcaatag attttgtctg ctgaaaaagt tcqatqaagc atttatccgc aaactaaggg ttcgtaattt gagccaataa tagttgatca aagccctgca ttacctatca aaaccgaaga acaaaccgat atcaagctat qaacctcaca atcatacagt gacgaaaaaa tagagcctgg ctagaagaat cgcgattttt tttttggcgc tacttgcagc gttatgatga ttcagattga ttttaagcta attttcttaa aaaattatcc ttaaacaaat gttatattat aattaagatc ccgaggaagc tttctttatt atggaaccgc tcaaaqggga caaaacccct gaaccaattt aattaaatta atatgaaatt gqtgatqaaa atttactggg agcaaaagag aaattttttt acaagaattt gattcgtaag aatttccgaa agttgttgac attaaatgca tgcgattacc tgtgactgaa cgataaccaa tatctcactt ttatggacaa ccaaaagaaa tgttattcgt aaattttgac tgattttata tgataattta cattgaacaa ccaatcagtt tttatcaaca ttctgctaat aacaaaaaaa taataattca aatttcttat 16 ctaaattttt ttgattatca gttaatactc aatgataaag caaaactcag cgcgatgttg cgaattatqg qggcttgaaa cgccttttta atcgatgttt gatgaaaaag qttgaacgtg tctgaaacgc aaacaaaacc atcgcgacaa cttgggcggc ctacgaggtc caggaccaac acaaatqgggg atcgaaggct caacagcgtg cattatatcc attttaccaa tcaagaatta cttaatgaat ttaaaggaaa cttgatagtc ttagttcagt ttcaacattt aatgctqaaa ccggcagata gcctaatcga gtgqtggcaa taattgctga gaatcgataa aaatagttca aatatattgt ctqgaagatc ttgaacctga aaaaattatc ataatatgcq atgaaatttt ttcataaaat tttcggagat aacttgaagc atatggctgg tttatattct gctctggacq tttttcggcg caattaaagg ttaacttcga atttaattta agaagatgat atcaagaaat ctcattttaa ttttaatcaa atcttqgaaa agtqacaaaa attcccagaa tagtagcaga aaatagaata gccctcaaga tqaagtagac aactaacctt agatttttat ggcaaatgct tcgaattcaa ccaggacggc ttattctgaa gacaaaaaca agggatgacc cgtgaatgtg tgccactagt ggggcagcca gctaaaccaa cgaaattatt aagaagaact tggaaccgat acaaggtgat ttttaccaat aaaatatatt tatgcgcaaa tgcccaaaga tattcgggct tcactataaa ctttgggcaa acaattagaa aacctacttt tcatattgac aaatccttat atccgcttat tatcaaagca aataattgat 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 WO 00/31115 WO 0031115PCT/AU99/01 035 caaataatcg cctctaacqa cagcctgaat ttattgaaaa gaggaatttg-aactttqaaa cctcttgatg aaaaacaaaa agcgaggaag aaaaaaacct atagaggaag aaattcaaaa gatcctgatg catataataa ggagagaatc acaacttgct aatcggzagat tattttagta aatacaagaa tgaaaatccc actgatattc gcggctgcaa aaactaaaaa agcaaactag aaaatgcagc caaaatttag caaaaagttg ggtgcggatt gaaaagcaaa ttgaaaaggt ttaacctaaa aggaacaact agattgtaga aatttqtgga atgcagataa agaagaaaaa tgaatcagga aaaggaaatq tgaaatgatg gattgaagaa ttttaaaaat ccat 2040 2100 2160 2220 2280 2340 2394 <210> <211> <212> <213> <220> <223> 12 798

PRT

Artificial Sequence Description of Artificial Sequence:Deduced protein sequence from clone pAD653 <400> 12 Thr Val Gly Ile Asn Lys Thr Glu Met Asp Ala Asn Thr Lys Arg Met 1 5 10 Met Tyr Asn Ala Asp Ile Thr Tyr Ser Val His Ser Glu Leu Gly Phe 25 Asp Tyr Leu Arg Asp Asn Met Val Phe Ser Ala Ala Glu Lys Val Gin 40 Arg Gly Leu Asn Phe Cys Leu Ile Asp Glu Val Asp Ser Ile Leu Ile 55 WO 00131115 WO 0031115PCT/AU99/O1 035 18 Asp Giu Ala Lys Thr Pro Leu Ile Ile Ser Giy Giy Lys Thr Asn Leu 70 75 Pro Ala Gin Tyr Leu Ser Ala Asn Gin Phe Val Asn Thr Leu Ile Ala 90 Glu Asp Phe Lys Gly Ile 115 Tyr Ile Asp Giu Giu Thr Lys Gly Ile Lys 100 105 Asp Lys Ala Asn Ala Phe Phe Giy Leu Arq 120 125 Leu Asn Asp 110 Asn Leu Tyr Giu Ile 130 Gln Asn Ser Giu Ile Vai His Arg Ile Gin 135 140 Arg Asp Val Giu Tyr Ile 155 Asn Ala Leu Arg Vai Gin Asp Gly 160 Ala 145 Asn Lys Val Met Lys Ile Ala Leu Asp Gin Phe Thr Gly Arg Ile Met Ala 170 Gly Arg 175 Ser Tyr Giu Ile Phe Phe 210 Ser Giu 180 Gly Leu Gin Gin Ala Leu Gin Ala Lys Giu Gly Leu .185 190 Giu Pro Giu Thr Lys Thr Leu Ala Thr Ile Thr Tyr Gin Asn 195 200 205 Arg Leu Phe Lys Lys Leu Ser Gly Met Thr Gly Thr Ala Lys 215 220 Thr Glu Glu Gin Glu Phe Ile Asp Val Tyr Asn Met Arg Val Asn Val WO 00/31115 PCT/AU99/01035 Ile Pro Thr Asn Lys Pro Met Ile Arg Lys Asp Glu Lys Asp Glu Ile 245 250 255 Phe Ala Thr Ser His Glu Lys Asn Gin Ala Ile Ile Ser Glu Val Glu 260 265 270 Arg Val His Lys Met Gly Gin Pro Ile Leu Ile Gly 275 280 Val Asp 290 Ser Glu Thr Leu Ser Glu Met Leu Asn Gin 295 300 Thr Ser Gin Val 285 Lys Gly Leu Tyr Ala Glu Ile Ile 320 His 305 Thr Val Leu Asn Ala Lys Gin Asn Gin Leu Glu 310 315 Ala Gin Ala Gly Arg Lys Asn Ala Ile Thr Ile Ala Thr Asn Met Ala 325 330 335 Gly Arg Arg Thr Asp Ile Ile Leu Glu Pro Gly Val Thr Glu Leu Gly 340 345 350 Arg Leu Tyr Ile Leu Gly Thr Asp Lys Ala Glu Thr Arg Arg Ile Asp 355 360 365 Asn Gin Leu Arg Gly Arg Ser Gly Arg Gin Gly Asp Val Gly Ile Ser 370 375 380 Arg Phe Phe Ile Ser Leu Gin Asp Gin Leu Phe Arg Arg Phe Thr Asn WO 00/31115 PCT/AU99/01035 Phe Asp Gin Ile Phe Gly Ala Tyr Gly Gin Thr Asn Gly Ala Ile Lys 405 410 415 Gly Lys Tyr Ile His Ala Val Leu Leu Ala Ala Gin Lys Lys Ile Glu 420 425 430 Gly Phe Asn Phe Asp Met Arg Lys Thr Val Leu Ser Tyr Asp Asp Val 435 440 445 Ile Arg Gin Gin Arg Asp Leu Ile Tyr Ala Gin Arg Asp Ile Leu Leu 450 455 460 Gin Ile Glu Asn Phe Asp His Tyr Ile Gin Lys Met Ile Ile Arg Ala 465 470 475 480 Val Asp Ile lie Leu Ser Tyr Asp Phe Ile Ile Leu Pro Asn Gin Glu 485 490 495 Asn Leu Ile Asn Phe Leu Asn Asp Asn Leu Ser Arg 505 510 Ile His Tyr Ile Thr Glu Gin 530 Lys Gin 545 His Phe Asn Phe Gly Gin Ile Gly Ile Glu Asn Tyr Pro Ile 515 520 525 Leu Asn Glu Phe Leu Ile Lys Gin Leu Glu Thr Ile Tyr Phe 535 540 Ile Gin Ser Val Leu Lys Glu Asn Leu Gly Lys Thr Tyr Phe 550 555 560 WO 00/31115 WO 0031115PCT/AU99/01 035 Glu Ser Giu Arg Tyr Ile Ile Leu Ser 565 Thr Leu Asp Ser Gin 570 Trp Gin 575 Asn His Ile Asp Thr Ile Asp Lys 580 Gin Tyr Ser Gin Lys Asn Pro Tyr 595 600 Leu 585 Arg Ser Ser Aia Asn Leu Vai 590 Giu Aia Thr Gin Ile Phe Thr Lys Lys 610 Phe Asn Ile Leu Val 615 Ala Glu Ser Ala Gin Ala Ile Val Ser 625 Leu Phe Asn Asn Ser Asn 630 Ala Giu Lys Giu Tyr Ile Lys Ile Leu Ser Asp Giy Thr Ala Ile Ser 645 Tyr 650 Pro Ala Asp Ser Pro Gin 655 Gu Ile Ile Ala Arg Lys 675 Asp Gin Ile Ile Aia Ser 660 665 Asn Giu Glu Arg Ile Ala Ala 670 Glu Lys Gin Ala Lys Giu Giu Gin Pro Glu Phe Leu Ala 690 Leu Trp 705 Lys Leu Lys Ile Giu 695 Lys Ile Giy Asp Ser 710 Lys Val Giu Ser Gly 700 Lys Leu Val Asn Leu Giu Giu Phe Giu Lys Lys Giu Met 720 WO 00131115 PCT/AU99/01035 22 Pro Leu Asp Giu Lys Gin Asn Ile Leu Vai Lys Met Gin Gin Giu Gin 725 730 735 Leu Giu Met Leu Glu Ile 755 Met Ser Giu Giu Giu Lys Asn Leu Ile Gin Giu Gin Asn 740 745 750 Vai Glu Ile Giu Giu Ile Giu Giu Giu Ile Gin Asn Giu 760 765 Asn Pro Gin Lys Val Giu Phe Vai Asp Phe Lys Asn Asp Pro Asp Aia 770 775 780 Tyr Asn Lys Leu Ile Phe Giy Ala Asp Tyr Ala Asp Asn His 785 790 795 <210> <211> <212> <213> <220> <223> 13 1410

DNA

Artificial Sequence Description of Artificial Sequence:Ciones pAD657 and pAD964 <400> 13 atgatttcqt atttttattc aaaatcagca ccqcagtgtc ttaaaactga aaatcccaqa ttttgttata aactaaataa aaatttagta aaatttcaaa aagaattaga tctattaaaa 120 cagaaaaaac ttqcaccaaa agaatatqaa agccagttta qtgatctaaa aqaaaaattt 180 ttagcttatg aagttaatat aaaaaaacat tatcaggcaa aaaaatccta taaattacqt 240 WO 00/31115 WO 0031115PCT/AU99/01 035 gctatttgag tttgaagcat gttgcccgga aatatcgtga ggcgaqtctg aacggggtaa tttcgtgctt ccgatgtcaa ttaaacaaga atgaccaaaa ggtatgattc atggatgaac cgcgatctcc gtcgcttctc acaagagatg atgcctgatg ttaaattcaa ttttttattg cttgatccaa tttgcaaaga atcggattca tttctaaaaa tcaaaaattt ttcgtaatgc gttcaggaaa ttgaatcagg gagaactttc ctttaaatcc ttgttaaaac ttgctaaccc aaagaattgt caacaacagc aaaaaagact ttqcaactta aaattctttt tcaataaagg ttgttggcqa aacctaaatt aagcaccaaa tgaggcaaga aaaatattga tgttgaatat aaacctttcc ctcaattgat atcagtaatt tqaaatcaaa aaattatcgg aacaaaaaaa gaaaaaagaa ggaaatggtt qatctcagca tttggataca aaaaattaca tatctcaatc aaatccaaga cgaacgactt tgcttttgca ttacagaata agttgtccca aaatggaaga catacaagct aaacaaatag tttgtcaatc ttttatgaag acttcttqtc ctttttaaca ggaaaagtta ataqgcatcc qcctatgaaa atgaaattat attttatcac accgtgcaag attattttca atqtatgctg catccatata cagtcaattc gttagaaacg agtccaactc ccaaaaatta ttaacagatc gaaataaacg cagcaaatcc gcgaaattca tttatggtct atccagt gca tttcagctgt aaattatgga aagcactttt atccccatga ttgaacctaa ctttagttct ttactcacga qtcaagttgt cttgagggct gcggggttgt attatgcctt accgagtcaa tttaccaaaa ccattttgat tcataaaatt cgagattaga tgccttaatt tgtcgqtcaa aaattttgat cttccagaat aggaatgtta atatottaga gctttcagga aattatcgtt tgatattatc ccttggagtt cgaggaaggt aattacttca tccttcttct agaacaagat atcagcttta atgactgcaa 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1410 <210> 14 <211> 470 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Deduced protein sequence from clones pAD657 and pAD964 WO 00/31115 WO 0031115PCT/AU99/01 035 <400> 14 Met Ile Ser Tyr Phe Tyr Ser Lys Ser 1 5 Giu Asn Pro Arg Phe Cys Tyr Lys Leu 25 Gin Lys Giu Leu Asp Leu Leu Lys Gin 40 Ala Pro Gin Cys Leu 10 Lys Thr Asn Lys Asn Leu Val Lys Phe Lys Lys Leu Ala Pro Lys Giu Tyr Giu Ser Gin Phe Ser Asp Leu Lys Giu Lys Phe 55 Val Asn Ile Lys Lys His Tyr Gin Ala Lys Lys Ser 70 75 Ala Ile Trp Asp Arg Ile Gin Lys Tyr Trp His Thr 90 Ser His Phe Asp Phe Giu Ala Phe Ser Lys Asn Val 100 105 Ile Gly Asn Lys Arg His Lys Ile Val Ala Ar Ile 115 120 Leu Ser Phe Val Asn Pro Ala Asn Pro Giu Ile Arg 130 135 140 Arg Asn Ala Ser Ile Asp Phe Tyr Giu Gly Glu Ile 145 150 155 Leu Ala Tyr Giu Tyr Lys Leu Arg Ser Phe Asn Arg Giu Tyr Lys Gin 110 Lys Asn Leu Asn 125 Asn Ile Vai Ile His Ala Leu Ile WO 00/31115 WO 0031115PCT/AU99/01 035 Gly Giu Ser Gly Ser Gly Lys Ser Val 165 Ile Thr Ser Cys Leu 170 Tyr Giy 175 Leu Val Gly Gin Asn Giy Val Ile 180 Asn Asn Pro Val Gin Asn Phe Asp 195 200 Tyr Arg Giy Lys Val Ile Ser Ala 210 215 Leu Asn Pro Thr Lys Lys Ile Gly 225 230 Ser Gly Giu Ile Lys Leu Phe 190 Leu Ser Asn Phe Arg Ala Trp Val Phe Gin Pro Met Ser Thr Ile Gin Ile 235 Met Giu Gly Met Leu Asn Lys Ile Val Lys Thr Lys Lys 245 Aia Tyr Giu Lys Ala Leu 255 Leu Tyr Leu Arq Met Thr Lys Ile 260 Leu Tyr Pro His Giu Leu Ser Gly 275 280 Ser Ala Ile Leu Ser Leu Giu Pro 290 295 Asn Pro Giu Met Val Met Lys 270 Ile Val Ile Giy Met Ile Gin Lys Ile Ile Vai 300 Arg 285 Met Asp Glu Pro Thr Thr Ala Leu Asp Thr Thr Val Gin Ala Leu Val 305 310 315 Leu Asp Ile Ile 320 WO 00/31115 Arg Asp Leu Gin PCT/AU99/O1 035 26 Lys Arg Leu Lys Ile Thr Ile Ile Phe Ile Thr His 325 330 335 Val Ala Ser Leu Ala Thr Tyr Ile Ser Ile Met Tyr Asp Leu Gly 345 Ala Gly Gin 355 Val Vai Glu Glu Gly Thr Arg Asp Giu Ile Leu Leu Asn 360 365 Giy Leu Ile Thr Ser Met Pro Asp Val 380 Pro Arg 370 Asn Lys 385 His Pro Tyr Thr Gly Giu Arg Leu Gin Ser Ile Arg Gly Vai Val Pro Ser Ser 390 395 400 Gly Asp Ala Phe Ala Val Arg Asn Asp Tyr Ala 410 415 Leu Asn Ser Ile Leu Giu Gin Thr His Arg 435 Phe Phe Ile Giu Val Lys Ser Ala Leu 440 Pro Lys Phe Tyr Arg Ile Ser Pro 425 430 Leu Asp Pro Lys Ala Pro Lys Val 445 Lys Trp Leu Gin Phe Ala Lys Met 460 Val Pro 450 Pro Lys Ile Ile Tyr Gin 455 *~Arg Gin 465 Giu Asn Gly Arq 470 WO 00/31115 PCT/AU99/01035 <210> <211> <212> <213> <220> <223> 27 1332

DNA

Artificial Sequence Description of Artificial Sequence:Clones pAD659 and pAD91O <400> atgaaaaata ttgaaaaaag tgaaataatt atctcccttg ttgatgttga taaagaattt ggtgataaaa cttttagggc tgaactactc aaacgtgata ggaaatattg gatgtagtct aagattaaag ttagatatta tcaaattgag cgaaagatca ctaactggaa gcacttgcaa gctttagata cttggcctaa aaaatcgcca gactatccag aaatacatta gagtttatcc ctaaattcag aagttttaga cctcagggtc gcggcgaaat cagctacaat aatttggtct ggaaaaaatt agttaaaaat aaaaacgtaa aaaattatct caaaacagga atgaaaatcg gatcgcttgt caaaaattag cttttatttt taatccgtaa ttaataaatg aaaaaaatca caggccaaaa gctattttaa tcaaataaat cgggaagaca caaatttaat ttttcaagat taaattaaaa tgagcactt a tttacaggct aaaattacag aacatccaaa aattaataaa agcaatttcg aattgagcct gcaaaaaatg tgttactcat tggaaaaatc ggtggctaat ggtcgaaatt attaqattgc aggaaaagtt ttqgacatta acaattttac gqcatcgata tatgcacttt agaattaaaa aagaaaaaat catttagaaa qataaattaq tcagaaagtt qaaattaccg gaattatcag gaaattgtcc caagtttttc gatcaagatg qcccaatacg tttattggtg cttggtctta ctgattcgtc atccaaaata aacgaggaga gtttaattgq taaaagacqt ttccacattt aaaaggcaga gacaggataa aactgcttga atgattctga ttaaaaaaaa atattattga gaggaatgaa tacttgatqa taaaaaaaat aagccttgca atgaaccaaa attctaattt aattatatac cagaagatat tttataaagg tt t tgt caca tggttttgaa tccggcacat atcagttcgt agaaattccg gcaaaagcga aaatccacag ttttagatat atacctaacc ccttgttggt acaacgcgta acctttatca tcaacaaaaa attatcaqat acaaatttat ttttcaggca aattcatgat cgatattgac ttcatactat 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 WO 00/31115 PCT/AU99/01035 28 tcacttgata tcaaagtaqa aaatacaata attaatqtcg aaactaacga tttttatgac 1260 ctcgagactc aagtttttct aaaatqagat gatgatgcta ttcatttaat qgagatqgaa 1320 aatgctgaaa tt 1332 <210> 16 <211> 444 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence:Deduced protein sequence from clones pAD659 and pAD910 <400> 16 Met Lys Asn Ile Glu Lys Ser Giu Ile Ile Ile Ser Leu Val Asp Val 1 5 10 Asp Lys Glu Phe Gly Asp Lys Lys Val Leu Asp Gin Ile Asn Leu Asp 25 Ile Lys Arg Giy Asp Phe Val Thr Leu Leu Gly Pro Ser Giy Ser Gly 40 Lys Thr Thr Ile Leu Arg Leu Ile Gly Gly Phe Glu Trp Thr Thr Arg 55 Gly Glu Ile Lys Phe Asn Gly Ile Asp Ile Lys Asp Val Pro Ala His 70 75 Lys Arg Asp Thr Ala Thr Ile Phe Gin Asp Tyr Ala Leu Phe Pro His 90 WO 00/31115 WO 0031115PCT/AU99/01 035 Leu Ser Val Arg Gly Asn Ile Giu Phe Gly Leu Lys Leu 100 105 Lys Arg Ile 110 Lys Lys Lys 115 His Leu Lys 130 Ala Glu Giu Ile Pro Asp Val Val Trp 120 Lys Lys Phe Giu 125 Lys Ile Lys Glu Lys Lys Trp, Asp Lys Gin Lys Leu Lys Ile Leu Gin Ala 145 150 His Leu Giu Lys Leu Giu Asn Pro Leu Asp Ile Lys Arg Lys Lys Leu Asp Lys Leu Asp Asp Ser 175 Asp Phe Arg Tyr 180 Ser Phe Lys Lys 195 Ser Asn Trp, Giu Tyr Leu Thr Ser Lys Ser Giu 190 Gin Giu Ile Lys Tyr Leu Arg Lys Ile Thr Asn Lys Giu Ile Thr Asp Ile Ile Asp Leu Val 210 215 Giu Asn Arg Ala Ile Ser Glu Leu Ser Giy Gly 225 230 235 Ala Leu Ala Arg Ser Leu Vai Ile Giu Pro Giu 245 250 Leu Thr Gly Asn Met Lys Gin Arg Val 240 Ile Val Leu Leu Asp 255 WO 00/31115 WOOO/1115PCT/AU99/0I 035 Glu Pro Leu Ser Ala Leu Asp Thr 260 Lys Ile Arg Gin Lys Met Gin Val 265 270 Phe Leu Lys 275 Lys Ile Gin Gin Leu Gly Leu Thr Phe Ile Phe Val 285 Lys Ile Ala Ile Thr His 290 Asp Gin Asp Giu Ala 295 Leu Gin Leu Ser Asp 300 Ile 305 Arg Asn Gly Lys Ala Gin Tyr Asp Asp Tyr Pro Val Phe Phe Gin Ala 340 Lys Trp Val Ala Glu Pro Lys Gin Ile Tyr 315 320 Phe Ile Gly Asp Ser Asn 335 Gin Val Glu Ile Leu Gly 350 Lys Tyr Ile Lys Lys Asn 345 Leu Lys Leu Tyr Thr Ile His 355 Asp Cys Leu Ile Arg Pro Giu 370 375 Asp 360 Giu Phe Ile Pro Gly Gin Lys Leu 365 Leu Asn Ser Gly Asp Ile Asp Ile Tyr Phe Lys Gly Lys Val 385 390 Ser Leu Asp Ile Lys Val 405 Ile Gin Asn Ile Tyr 395 Giu Asn Thr Ile Ile 410 Lys Gly Ser Tyr Tyr 400 Asn Val Glu Thr Asn WO 00/31115 PCT/AU99/01035 31 Asp Phe Tyr Asp Leu Glu Thr Gin Val Phe Leu Lys Trp Asp Asp Asp 420 425 430 Ala Ile His Leu Met Glu Met Glu Asn Ala Glu Ile 435 440 <210> <211> <212> <213> <220> <223> 17 1590

DNA

Artificial Sequence Description of Artificial Sequence:Deduced protein sequence from pAD662 <400> 17 gatattaaat cctqtaattg qttggtgaaa gctggacaat aaagcctccg ttatcaattt tgaaataaga ccaacaacaa gcactttcaa gaaaatgctt atttttgtct cgcgatggta ctaattcagg tgaaaaaaac ctctttctga atggcqcggg atgaaggtga aacgtgtcgg qcgagaacat tgatttccga ttgctggctc aaaatgcaaa ttcgcttact cccataaatt aattcatttc acattgttqq taatattcta tgtgactttt aaaatcgaca tctaattttt aattgcaata gtatatcggt atgcaaaaaa tctttcgatt actactaatt tgatattatq aaatqaagtc ccagtataat ccggccttta tcactaaaaa gttqttccaa cttttaaaaa gaagataaaa attcatcaag aattatccga tatctagaaa gcaaaacagc ttagatgaac aaaagtttaa aaatatgtct aaaaatqaag aagtccaaat aaataaaaaa aagqggaaat ttttatcagg ttatggcttt aactttcaat ctaaatttgq tggtcggtct aaatggttqa cgacttcctc aaagtaaaqq ctqataatat aaataattga ttcctcctag agtttatgqt aactagccta agtgattcct tgcaaataca ttcaccttat caaagttaac tgatgaagat gatcgcaaaa tttaaatgat aattacttcg tgttgtaatc tgaaaaccgg ggatttagat WO 00/31115 WO 0031115PCT/AU99/01 035 cgaaaaatcg gcaaattata atttccggac tataacaaac acaaaacagg ctaatccaaa aaatgaggaa gtaagtatta caaaaagttg ccaacaaaag tcatcacaag accgatcacc cagtttttgc attttqagac gggaaattat atgttgtcaa ttgttggatc cttcaagtqg caatcaaatc ttttttcgat ttctaaataa aaacaaaaaa taattgccaa gtattgatgt gtaaaacaat tttattttgt atctttttta ataatcttgg ttttgagatc aaatgcttcc gggtcgaacc caaagttttc gggaattatg tcaaaataat aaataaggaa tattctaaat agctttaagc tggctcaaaa tattgtaatc gactgqaaaa taactttttt tgaagcgagt aaagatatag cttgatgtta gaattaatgc tataaagqta tatgcttccg atcacttccg ataattaatg aatgtcgaat accaaatttg tacgaaattt tcttcggaaa ctttttggtc gaccaaaagg ttattcctca aacaaggcqa tttcactttt aagaagtaaa aagatcgaaa ccgctttgca cccaaaaact ccctttctgg accttctaat ataaaatttt ttqaaqaact aaaatcaaga attaccataa tgctagtatt aattgtcgga tgggcagtat atttactaac aaatgttggt tttattttca aaaaaaagat gggaaatcag tatcgatgag actagacctt tttaggaatc ctgactttcc ggacttccaa 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1590 <210> <211> <212> <213> <220> <223> 18 530

PRT

Artificial Sequence Description of Artificial Sequence:Deduced protein sequence from clone pAD662 <400> 18 Asp Ile Lys Leu Lys Lys Thr Asn Ile Leu Ser Leu Lys Lys Ile Lys 1 5 10 Lys Val Tyr Gly Pro Val Ile Ala Leu Ser Asp Val Thr Phe Val Val 25 WO 00/31115 PCT/AU99/01035 33 Pro Lys Gly Giu Ile Thr Ser Leu Val Giy Giu Asn Gly Ala Gly L ys 40 Ser Thr Leu Leu Lys Ile Leu Ser Giy Val Ile Pro Ala Gly Gin Tyr 55 Giu Gly Asp Leu Ile Phe Glu Asp Lys Ile Met Ala Phe Ala Asn Thr 70 75 Lys Ala Ser Giu Arg Val Gly Ile Ala Ile Ile His Gin Giu Leu Ser 90 Ile Ser Pro Tyr Leu Ser Ile Cys Giu Asn Met Tyr Ile Gly Asn Tyr 100 105 110 Pro Thr Lys Phe Gly Lys Vai 115 Lys Lys Tyr Leu Giu Met Val 130 135 Asn Trp Asn Lys Met Ile Ser Giu Cys 120 125 Gly Leu Asp Giu Asp Pro Thr Thr Ile 140 Ala Gly Ser Leu Ser 145 Ala Leu Ser Lys Asn 165 Ser Leu Asn Asp Giu 180 Ile 150 Aia Lys Gin Gin Met Vai Gu Ile Ala Lys 155 160 Ala Lys Leu Leu Ile Leu Asp Giu Pro Thr Ser 170 175 Asn Ala Phe Arq Leu Leu Asp Ile Met Lys Ser 185 190 WO 00/31115 PCT/AU99/01035 34 Leu Lys Ser Lys Giy Ile Thr Ser Ile Phe Val Ser His Lys Leu Asn 195 200 205 Giu Vai 210 Lys Tyr Val Ser Asp Asn Ile Val Val Ile Arg Asp Gly Lys 215 220 Phe 225 Ile Ser Gin Tyr Asn Lys Asn Giu Giu Ile Ile Asp Giu Asn Arg 230 235 240 Val Gly Arg Pro Leu Lys Ser Lys Phe Pro Pro 250 255 Leu Ile Gin Arg Asp Leu Ile Val Ile 275 Asp Asp Arg Lys Ile Gly Gu Ile Ile Phe Giu Ile Lys Asp 260 265 270 Pro His Ala Ser Ile Ala Asn Tyr Asn Val Val Lys Asn 280 285 Ala Ser 290 Leu Asp Vai Lys Gin Gly Giu Ile Val Giy Ile Ser Gly Leu 295 300 Glu Leu Met Leu Ser Leu Phe Giy Gin Tyr 315 320 Val1 305 Giy Ser Giy Arg Tyr Asn Lys Pro Lys Phe Thr Asn 340 Ser Ser Gly Lys Vai Phe Tyr Lys Giy Lys Giu Val 325 330 335 Thr Lys Gin Ala Ile Lys Ser Gly Ile Met Tyr Ala 345 Ser Giu Asp Arq Lys Asn Val Gly Leu Ile Gin Ile Phe Ser Ile Gin WO 00/31115 WO 0031115PCT/AU99/01 035 Asn Asn Ile Thr Ser Ala Ala Leu His Leu Phe 370 375 Ser 380 Lys Trp Gly Ile Leu 385 Asn Lys Asn Lys Giu Ile Ile Asn Ala 390 Lys Leu Lys Lys Val Ser Ile Lys Thr 405 Lys Asn Ile Leu Asn 410 Asn Val Glu Ser Leu Ser 415 Gly Gly Asn Phe Asp Leu 435 Gin Lys Val Val Ile 425 Ala Lys Ala Leu Ser Thr Lys 430 Asp Val Giy Leu Ile Ile Asp Giu Pro 440 Thr Lys Gly Ile 445 Ser Lys 450 Tyr Giu Ile Tyr Lys Ile Leu Leu Asp Leu 455 460 Ser Ser Giu Ile Giu Glu 475 Lys 465 Thr Ile Ile Val Ser Ser Gin Gly Leu Leu Gly Ile 480 Gly Gin Asn Gin 495 Phe Phe Asp Gin 510 Thr Asp His Leu Asp Trp Leu Ser 500 Lys Asp Tyr His Tyr 485 Phe Val Thr Gly Lys Leu Phe 490 Gin Phe Leu His Leu Phe Tyr Asn 505 Lys Asp Phe Gin Ile Leu Arg His AsnLeGlGu Leu Gly Glu WO 00/31115 WO 0031115PCT/AU99/01035 Ala Ser 530 <210> 19 <211> 852 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence:Clone pAD681 <400> 19 atcgaaacca ttagtcgaaa ttcggaattt gaaaaaccat tcccatatta accttggttg agtgaatttg gaaattgata gatccagaaa ctaaaacaga gaagaaatca gaaatgacga gcaattgtcg tttqttcttg ggtcgacctg tcaaaattga aacaacgact gttcaggaat ttagtttact tgatcagcca actatggtta agaaatatca aaacaaatgg tagtaattga aggaaaaaaa gcaagtatct cttttgagcg ggataaattt aagccggtct ca gctgggtgag aaaagaccga tgagaattat aqattattta aattaagggc tcqactacct acaggcaaaa cgaaattgtc atctaccaaa agaaagaagt ccaqgaagaa cgatgaaatc acttcgtgaa tgtttctttt cagctcgaat gtcaataatq gcccgccata ccccaagacg atqtatqaag tcagctcttg atttatgttt tcgcaaatta aatcaaatqq ctcattttaa aaattacaaa ataattaagc ggcgttgata ttgcgatccg tttttgaaaa atattgactc ttgacggacg gcctaatttt gtqatcgaag ataatcggcc cagccacacc tcagaccaac agaaaatttt ctaccacaaa atgtctatta ttcgaaaagg tcccggaagt catgcgagct aaataataaa gctttctgaa ccaaaaaggc tattgatgaa ccgaaaacaa cttaaaactc ggccagctat tggactaatt tcagtatttg acgactggcc tttgcactca aatttatgat ttctttgatt cggtaccccg WO 00/31115 PCT/AU99/01035 <210> <211> <212> <213> <220> <223> 284

PRT

Artificial Sequence Description of Artificial Sequence:Deduced protein sequence from clone pAD681 <400> lie Glu Thr lie Lys Ile Glu Leu Gly Glu Gin Leu Glu Phe Phe Glu Lys Asn Asn Lys Leu Val Glu Lys Gin Arg Leu Lys Asp Arg Val Asn 25 Asn Asp Ile Asp Ser Leu Ser Glu Phe Gly Ile Cys Ser 40 Gly Ile Glu Asn Tyr Ser Leu Ala Arg His Ile Asp Gly Arg Gin Lys Gly Glu 55 Lys Pro Phe Leu Asp Tyr Ser His Ile Met lie Ser Arg Lys Gin Thr 100 Leu Pro Gin Asp Gly Leu Ile Phe Ile Asp Glu 70 75 Ser Gin Ile Lys Gly Met Tyr Glu Gly Asp Arg 90 Leu Val Asp Tyr Gly Tyr Arg Leu Pro Ser Ala 105 110 WO 00/31115 PCT/AU99/01035 38 Leu Asp Asn Arq Pro Leu Lys Leu Ser Giu Phe Giu Lys Tyr Gin Gin 115 120 125 Ala Lys 130 Ile Tyr Val Ser Ala Thr Pro Ala Ser Tyr Giu Ile Asp Lys 135 140 Thr 145 Asn Gly Giu Ile Val Ser Gin Ile Ile Arg Pro Thr Gly Leu Ile 150 155 160 Asp Pro Giu Ile Val Ile Giu Ser Thr Lys Asn Gin Met Giu Lys Ile 165 170 175 Phe Gin Tyr Leu Thr Thr 195 Leu 180 Leu Lys Gin Lys Giu Lys Lys Glu Arg Ser Leu Ile 185 190 Giu Giu Ile Ser Lys Tyr Leu Gin 205 Thr Lys Arg Leu Giu Giu 210 Phe Giu 225 Lys Leu Gin Asn Tyr Tyr Leu His Ser Giu Met Thr Thr 220 Arg Asp Giu Ile Ile Lys Leu Ala Ile Vai Gly Ile 245 Val Ser Leu Ile Phe 260 Asn Leu Leu Arg Giu 250 Val Leu Giu Aia Gly 265 Arg Lys Gly Ile Tyr Asp 235 240 Gly Val Asp Ile Pro Giu 255 Leu Val Ser Phe Leu Arg 270 WO 00/31115 39 Ser Ala Cys Glu Leu Gly Thr Pro Gly Arg Pro Ala 275 280 PCT/AU99/01 035 <210> <211> <212> <213> <220> <223> 21 843

DNA

Artificial Sequence Description of Artificial Sequence:Clone pAD700 <400> 21 atgcccaaat acaaatgtaa attgttcaag tctaagaagg ggatcaaatt caacttcaga ataaattact gaaccaattt aaaattttta aatttatttq ccaaatqcta aaacttttta gcactqaaaa ccagccatga gt taaaccgact tggaaatgat aatattttga ttgaggaatt taggtttttg aaaatgatga ctaatttaat tagaaatctc tttgttataa atttcaaaaa ctgaggtttt tcgaatccaa tgccaqttga ttaccccaga tagagcccga tgcgaatgca aaatttagat aactaatgct tggtgccctt aattatcgtc tattaaattt aaactttgta caaatttatc aaatactata taaaaaatta attagttgag aattttgcat aattattgag tttgtccaga aaaattccaa tatatttttc gattccaaaa aataatttaa tttgqtgaaa tttctaaata aatcaatcga agtattatac aaatacggtg atgccctttt acttcaacta aagtttagaa attattagtq ttcaaaacat aaataaaaaa aaaatattct aaaatcttta tcttaaaaaa aaatttataa ttgaagaaac tttttgagag attcaagccc gttacgqgat atataaaatc gacgaacatc acagaaataa gtaaaatgtt tgaaaaaatg c a agt t taa a tgcaaattta tattattttt tgttgtccca ttttttgtca taattttagt aaaatataaa agagatgcaa tgagtttgaa catccttgaa aatggaaagt ttccagtcgt qaaaaagtta 120 180 240 300 360 420 480 540 600 660 720 780 840 843 <210> 22 WO 00/31115 PCTIAU99/01 035 <211> 281 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence:Deduced protein sequence from clone pAD700 <400> 22 Met Pro Lys Leu Asn Arg Leu Arg Ala Arg Phe Val Gin Ile Gin Asn 1 5 10 Ile Glu Lys Met Thr Asn Val Met Glu Met Ile Ala Asn Ala Lys Ile 25 Pro Lys Ile Lys Asn Lys Phe Lys Ile Val Gin Glu Tyr Phe Glu Asn 40 Leu Asp Tyr Ile Phe Gin Asn Ile Leu Ala Asn Leu Ser Lys Lys Val 55 Glu Glu Leu Thr Asn Ala Asp Ser Lys Lys Asn Leu Tyr Ile Ile Phe 70 75 Gly Ser Asn Leu Gly Phe Cys Gly Ala Leu Asn Asn Leu Ile Leu Lys 90 Asn Val Val Pro Gln Leu Gin Lys Asn Asp Glu Ile Ile Val Phe Gly 100 105 110 Glu Lys Ile Tyr Asn Phe Leu Ser Ile Asn Tyr Ser Asn Leu Ile lie WO 00/31115 WO 0031115PCT/AU99/01035 125 Giu Pro Ile Leu Lys Phe Phe Leu Asn Ile Giu Giu Thr Asn Phe Ser 130 135 140 Giu Ile Ser Asn Phe Val Asn Gin Ser Ile Phe Glu 145 150 155 Arq Lys Tyr Lys Ile Phe Ile Cys Tyr Asn Lys Phe Ile Ser Ile Ile His Ser Ser 165 170 175 Pro.Glu Met Gin Asn Leu Phe Asp Phe Lys Lys Asn Thr Ile Lys Tyr 180 185 190 Gly Giy Tyr Gly Ile Giu Phe Giu Pro Asn Ala Thr 195 200 Lys Leu Met Pro Phe Tyr Ile Lys Ser Ile Leu Giu 210 215 220 Giu 205 Val Phe Lys Lys Leu Phe Ile Giu 225 Ser Lys Leu Vai Giu Thr Ser Thr Arq Arg Thr 230 235 Ser Met Giu Ala Leu Lys Met Pro Val Giu Ile Leu His Lys Phe Arg Asn Arg Asn 245 250 255 Asn Ser Ser Arg Pro Ala Met Ile Thr Pro Gu Ile Ile Gu Ile Ile 260 265 270 Ser Gly Lys Met Leu Lys Lys Leu Gly WO 00/31115PC/U9003 PCT/AU99/01035 <210> <211> <212> <213> <220> <223> 23 615

DNA

Artificial Sequence Description of Artificial Sequence:Clone pAD7l1 <400> 23 tcaggtatgt ggaattatgc tttcttgata cacccaatcg ttacaacaat cctgggattc tcagttttca ttaggtcttc acacttgaaa agtaaaaaga caggatcttt caaaaaatat ctgtaatttt gaaatacctc gtcttatcat cccgagttga gcccaggaga gtgcgattta cttcggcaat cgatttcgca tccgcaaaaa tatgg taaggaaatt tcaatcctac tqccttaatt atcgtttcac agcagttcgg -aattgaatag ttttattgtt tttaatgtct taaaattgca caggattttq acagactgaa gattatttaa tcttaccttt ttaggaattc cacaatttca gqaacttcga gatcaaaqcc cgttatgatg tttacaa gtt cgaaaaaatt cccttaaatt tgccgacact ataataatat ccttttcaat ccaaaaattc tttcagtggt tccaacctgt taataatttt cacaaaaagt ctccttctat aaatcctgct ttttagcgga gcaaatttat aataatgtcc aacttttatc tttgcgactt tqaaattatg agcaacaact tctaaaacaa tgattcaaat 120 180 240 300 360 420 480 540 600 615 <210> 24 <211> 205 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Deducedt protein WO 00/31115 PCT/AU99/01035 43 sequence from clone pAD711 <400> 24 Ser Gly Met Ser Lys Asn Ile Lys Glu Ile Ser Ile Leu Pro Leu Lys 1 5 10 Leu Asn Pro Ala Gly Ile Met Pro Val Ile Phe Ala Leu Ile Ile Val 25 Ser Leu Pro Thr Leu Phe Ser Gly Phe Leu Asp Arg Asn Thr Ser Ala 40 Val Arg Asn Trp Ile Asp Asn Asn Met Gin Ile Tyr His Pro Ile Gly 55 Leu Ile Ile Phe Ile Val Phe Asn Val Ser Phe Ser Ile Ile Met Ser 70 75 Leu Gin Gin Ser Arg Val Asp Lys Ile Ala Gin Asp Phe Ala Lys Asn 90 Ser Thr Phe Ile Pro Gly Ile Arg 100 Leu Ile Ser Val Val Leu Arg Leu 115 120 Thr Phe Leu Gly Ile Leu Gin Pro 130 135 Pro Gly Glu Gin Thr Glu Asp Tyr 105 110 Ser Val Phe Ser Ala Ile Tyr Leu 125 Val Glu Ile Met Leu Gly Leu Pro 140 WO 00/31115 WO 0031115PCT/AU99/01 035 44 Ser Ala Ile Thr Ile Ser Gly Thr Ser Ile 145 150 Ile Ile Leu Ala Thr Thr 155 160 Arg Tyr Asp Ala Gin Lys 175 Thr Leu Giu Thr Ile Ser Gin Ile Lys 165 Val Leu Lys Gin Ser Lys Lys Ile Arg 180 185 Asn Ser Pro Ser Ile Asp Ser Asn Gin 195 200 Lys Asn Leu Gin Val Arq Lys 190 Asp Leu Leu Trp 205 <210> <211> <212> <213> <220> <223> 648

DNA

Artificial Sequence Description of Artificial Sequence:Cione pAD721 <400> atcgccgaag gatactgcaa gtactaagtt attgtcgaac gacattqata gcttttatgt gaaaaaatta caaaacaaga actcagatca aagtttcaag ctgcaggtga t accaaatga ttcttgatga ctcgtaaaca cagttgatat gaaactataa aaataatcgg aagaaattgt tttttctccg tattttcacc gctaatcaat agtccctgat aataaatcaa cgtctatctt agatatctcc ggcaactact ttttcaagtc tttgaccgac tacttttcac aaattacttg aatgttgcca cttttgcagt tttaaagatt gaattagtgc ttagaagata ccttttcttg ttttattttt cagaaattca atgaacttta aaagaatttt ataccqacqa cgactt gtca attattatta ttgtcttttc ttacactgat taggatgtta gaccaaatta tgttgatgaa gcgcaacatt tcaaattggc tcaagcactt tgtcgttgat tgatcctaat aaaaaagatt PCT/AU99/01035 WO 00/31115PC/U9003 atqccgccga agtttttgcc caaaatgatt tttccagtac tcccggttgt taataccaqc 600 cagaaactaa tccgaatggt tccagttgat gatattatcc qatattgt 648 <210> <211> <212> <213> <220> <223> 26 216

PRT

Artificial Sequence Description of Artificial Sequence:Deduced protein sequence from clone pAD72l <400> 26 Ile Ala Glu Glu Val Ser Ser Phe Ser 1 5 Pro Phe Asp Arg Leu Leu Phe 10 Phe Arg Met Ser Pro Glu Ile Asn Lys Leu Asp Thr Ala Thr Ala Gly Asp Ile Phe Thr Tyr Phe 25 Val Leu Ser Leu Pro Asn Glu Leu Ile Gin Thr Lys Leu 40 Leu Leu Asp Leu Asp Glu Val Pro Asp 70 Asp Ile Asp Thr Arg Lys Glu Leu Tyr Val Asp Glu Ile Val Glu Leu 55 Asn Val Ala Lys Arg Ile Leu Arq Asn Ile 75 Gln Ile Asn Gln Leu Leu Gin Tyr Thr Asp 90 WO 00/31115 PCT/AU99/01035 Asp Gin lie Gly Ala Phe Met Ser 100 Asp Ser Thr Cys His Gin Ala Leu 115 120 Val Asp Ile Val Tyr Leu Phe Lys 105 110 Glu Lys Ile Arg Asn Tyr Lys Asp 125 Ile Ser Glu Leu Val His 130 Ile Ile Gly Ala Thr Thr 145 150 Tyr Tyr Val Val Asp Gin Asn Lys Lys 140 Leu Glu Asp lie Phe Ser Asp Pro Asn 160 Thr Leu 175 Thr Gin Ile Lys Glu Ile 165 Val Phe Gin Pro Phe Leu Val lie Lys Lys Val Leu Pro 195 Val Asp Asp 210 Ile Met Pro Pro Lys 180 Val Val Asn Thr Ser 200 Ile Ile Arg Tyr Cys 215 Phe Leu Pro Lys Met Ile Phe Pro 185 190 Gin Lys Leu Ile Arg Met Val Pro 205 <210> 27 <211> 837 <212> DNA <213> Artificial Sequence WO 00/31115 47 <220> <223> Description of Artificial Sequence:Clone pAD727 PCT/AU99/01 035 <400> 27 tcatttgqtt gaaagctcaa tttcttgatt qaaaatggaa gcctcattaa atttcgaatg cttatttttg attttctcac gaaatatcgc acaatcgtgg tataaatttg tcggtatata tcaattgata caggatttaa attcaagtta cacaaaatat atatttttaa atacggcaat aagaagctca taactatttt ttctttttaa agaatttagt tttttgatag ataaaattaa catccttgac ttgtctttaq tttagcaatt tgatttaatt aaacaagtca tcttqaaatt ccagaataat aaaattaatt tacgctaatt tgttttaatg tgttgcaatt agtaaaagca agaaatttcc aaccatttta cctgacgatg gattttagtg acaaaggaaa aattcaacaa aaaacaaaac ttttctaata ttcaatgcaa aggtttaaat gttttgctag cttactttaa aqattttcag ttacaagcaa acaattgttg ttaattggtg gcggaactaa aagccgaaaa ttttgcttaa ttqatctagc ttcqaatgac ttctttcagt ggactttttc caattattat ttggttatac aaataaatca ttagagaaac ttttaatgat tgataattgg ctttttaatt aataattagt tccattaaat aacaaaaatt aaattattca tggaatcgca qcttqcaata tacacggatc agtaaatgat tgaaaatgtt agcaaaaagg tftttatgaa aacatattcc 120 180 240 300 360 420 480 540 600 660 720 780 837 tccttattta tcgcaacccg catttggatt attcttgaat catcccgtaa tcgtaaa <210> <211> <212> <213> <220> <223> 28 279

PRT

Artificial Sequence Description of Artificial Sequence:Deduced protein sequence from clone pAD727 <400> 28 Ser Phe Gly Ser Gly Phe Asn Leu Ala Ile Asp Phe Ser Gly Gly Thr WO 00/31115 WO 0031115PCT/AU99/01 035 Asn Phe Leu Ile Giu Ser Ser Asn Ser Ser Tyr Asp Leu Ile Thr Lys 25 Glu Lys Ala Glu Lys Ile Ile Ser Phe Leu Asp Ser Gin Asn Ile Asn 40 Lys Ser Asn Ser Thr Ile Leu Leu Asn Pro Leu Asn Glu Asn Gly Asn 55 Ile Phe Asn Leu Glu Ile Lys Thr Lys Leu Asp Leu Ala Thr Lys Ile 70 -75 Ala Ser Leu Asn Thr Ala Ile Gin Asn Asn Phe Ser Asn Ile Arg Met 90 Thr Asn Tyr Ser Ile Ser Asn Giu Giu Ala 100 105 Gin Lys Leu Ile Phe Asn 110 Ile Phe Thr Ala Ile Leu 115 Ser Val Gly Ile Ala Leu Ile Phe Vai Thr 120 125 Leu Ile 130 Arq Phe Lys Trp Thr Phe Ser Leu Ala 135 Leu 145 Phe Asn Val Leu Met Val Leu Leu Ala Ile 150 155 Ile Ile Phe Ser Leu 140 Ile Ile Thr Arg Ile 160 Thr Leu Ile Gly Tyr 175 Giu Ile Ser Gin Asn Leu Val Vai Ala Ile Leu 165 170 WO 00/31115 WO 0031115PCT/AU99/01 035 Thr Val Asn Asp Thr Ile Val Val 180 Ser Glu Ile Asn His Giu Asn Val 195 200 Ile Ser Leu Gin Ala Ile Arg Giu 210 215 Asp Arg Val Lys Ala Arg Phe 190 Ile Lys Glu Tyr Lys Phe Asp Thr Ala Lys Arg 220 Ser Val Tyr Thr Ser Leu Thr Thr Ile Leu Thr Ile Val Val Leu met 225 230 235 Ile Phe Tyr Ser Ile Asp Ile Val Phe Ser Leu Thr Met Leu Ile Gly Val Ile Ile 245 250 255 Gly Thr Tyr Ser Ser Leu Phe Ile Ala Thr Arq Ile Trp Ile Ile Leu 260 265 270 Giu Ser Ser Arg Asn Arg Lys 275 <210> 29 <211> 2523 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence:cione pAD742 WO 00/31115 WO 0031115PCT/AU99/01035 <400> 29 atgaagttaa aaatttgatg aatgatgcaa atggatgatg atcaggacaa aaaatttacc gatattaaaa tttcaagaac acacaaaaaa ctaaaagagc atgattttat aaatatagtc ctcaaaaaac gaaaggaatt ctaaaaaagt aatagctttc aaatatacag caacctgaaa acatttatcc ctaaattcac tacaagattt qcaaatcgaa qttattaqaa tcaattcagt caagaattta ttacagcaaa gaattaatcg gacctaaatt ttagaacttq taaaaattga gttcaattgt ttaaatgagt gcatttttgc aagaqcttga tcgaaaaatc agtaccaaac ctaaatttat ggtcagaaat aaattaatac caagtgaaat ttataagcgc tagaaattga tatacagtct ccttagaatc tttttcctgg gcgcaattga ccgcagtttc ccctttcaac aaaaaggttt taqcggattt tcgcaaactt ctagcgggat ataaaatcqa aagttaaatc ttaatattaa aaattaaagc taagtctaaa aaaatcaatq aattgaaggt tqgaataatt tttaggcgaa tqgctcaaaa aaaacaagta aaagcaatta agaattagat cagcgaaatt tgaagctgaa cctaaattta tagtgtaaat gcaaaaagac agaaaaactt aaattccgaa aatgtctttt atattgtqgq agccgcattg agcaattaat attaaaaccg tataaattta tttactagga ccttaaccac aatttctggg tqaactaaca taatgaattt tcttaatqat tcaaaatgaa tcqaacttta caaaatttta tttaaatcct gggccaaatg aaatcagtta acaggtatga aaacagcttg gaatctgttg caagtccaqg gaagcaaatg atcaacacta qcttatgcaa tttgagcaaa aattttgcaa aatacattta aaaattatta ttgccaaagg cttgtctctg ggtccaactt tttctaaaqa cgatttattc gctagcaatt ggqattattg aaaaacatga ggtcataaga aataatttga gaacagttaa ttagagcaaa ggtcttgaaq aaagaaaaaa aaaactgaaa ttgctgaacc gctctqgaaa aacaattacc tcgcaattga aaaaacaagc aagtcggctt aaaaattaaa agaaaataat aaaaccgcqa aagcaactca aaatggctgc aattaatcaq ggacacaaaa gtcaaactag ggacaaaaat atttaatcaa taaaacaaat aaaattatgc ccgatttata tagtcgattt ttgctgatac tcgtaacttt taaaaaatga atttttttga ttacaagaga aattt agtaa agagtctaaa ttgaacttga aaaaacagct tgtaagtatt atccaatata gggcccaaat aaaaattgaa agaaaatgca aattgtctct tgatctaaaa tattacaaat aattcaccgc acttcaagaa tttgcgccaa ccaaaacaag gggtgaaatt aatctccgag cattatcgaa aattttccca tgttgttgac aggaagcgca tcttgaacat tgaaaaaaaa aattgattca agacggcgat ttcttccttt agaaaaaatt atctgtattt ttctgaaaat tcaaaaagat aaatgtggtt cgataatcaa 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 WO 00/31115 WO 0031115PCT/AU99/01 035 atttcagaac gatcttaacc aataatttat qaaattccag cttggaaatg aaactaaaaa tcggatttag gagtttaacc gataaaaacg atcaaaaata ttttcaatta cttgatqaqt cagtttttga at c ttacaqtttt aaaatcaaaa cccaaaaata aaaaagatqc ttaatttaga aaagtcaaaa ataaaattat ttgttttcca atattttaaa ttaggctttt ttaaggcaaq caaatgtgat tcatcaccca agttcaagaa ttacaaagac cagtctaact tcgcgaattt tgcaattaat tgaacttgaa cattgqaaaa gaatatgttt ttcggggatt ttctgqqggc accaattccg cagatatgtg tcggcacgga cttaaccaaa aaatatgaat tttgaaggcg qttaataqtc gactttgaaa actgccaggt acccaqgaaa ggtgggggaa qaaataagtg gaaaaggcaa ctttgcattc qaatttcgaa acgatgtccc aacagcgaaa ttttaattac cagctcaaaa taaatttaga caacgagtca caaaaatttt ttgtcgatct gtgcaaaaat cccaaccacc ttattgcaat ttgatgaagt agcagttaaa gagttqatca aatcaacgca aaatttacga atatqaactt gattaaagcg aagactcgaa agaagttatc agttaattcc ttattttagt tggaaaaact ttcacttttg tgaagctgcc acaaaaaacg acttttagga 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2523 <210> <211> <212> <213> <220> <223> 841

PRT

Artificial Sequence Description of Artificial Sequence:Deduced protein sequence from clone pAD742 <400> Met Lys Leu Ile Lys Ile Glu Ile Glu Gly Phe Lys Ser Phe Ala Glu 1 510 Pro Val Ser Ile Lys Phe Asp Gly Ser Ile Val Gly Ile Ile Gly Pro 25 WO 00/31115 PCT/AU99/01035 52 Asn Gly Ser Gly Lys Ser Asn Ile Asn Asp Ala Ile Lys Trp Val Leu 40 Gly Giu Lys Ser Val Lys Gin Leu Pro Gly Pro 55 Ile Phe Ala Gly Ser Lys Thr Gly Met Ile Ala 70 75 Asn Met Asp Asp Gly Ile Glu Lys Ile Giu Lys Gin Ile Arg Thr Lys Giu Leu Giu Lys Gin Vai Lys 90 Gin Leu Giu Ala Giu Asn Lys Ile Tyr Leu Giu Lys Ser Lys Gin Leu Giu Ser 105 110 Val Giu Vai Gly Leu Ile Val 115 Asp Ile Lys Lys Tyr 125 Gin Thr Giu Leu Asp Gin 130 Val Gin Giu Lys 135 Leu Asn Asp Leu Lys 140 Phe Gin Giu Pro Lys Phe Ile Ser Glu 145 Thr Gin Lys Arq Ser 165 Gl Ile His Arg Leu 180 Giu Ala Asn Giu Ile Ile Ile Thr Giu Ile Giu Ala Glu 170 Lys Glu Gin Ile Asn 185 Ile Asn Thr Lys Asn Arg 175 Thr Leu Asn Leu Ala Tyr 190 WO 00/31115 WO 0031115PCT/AU99/01035 Ala Lys Ala Thr Gin Leu Gin 195 Val Asn Phe Giu Gin Lys Met 210 215 Giu Met Ile Leu Ser 200 Ser Giu Ile Ser 205 Lys Tyr Ser Leu Ala Ala Leu Arg Gin 220 Ile Ser Ala Gin Lys Asp 225 230 Leu Lys Lys Leu Giu Ile 245 Asn Phe Ala Lys Ile Ser Gin Asn Giu Giu Lys Leu 250 Asn Thr Phe Arg Thr Gin 255 Lys Gly Giu Ile Giu Arg Asn Leu 260 Ile Ser Gin Thr Arg Ile Ser Giu 275 280 Tyr 265 Ser Leu Asn Ser Giu Lys Ile 270 Giu Ser Met Leu Lys Lys Ser Leu 285 Ser Phe 290 Leu Pro Lys Gly Lys Ile Ile Ile Asn Ser Phe Leu Phe 305 Pro Gly Tyr Cys Gly 310 Leu Vai Ser Asp Giu Ala Ala Leu -330 Ile Lys Ile Phe Lys Tyr Thr Gly Ile Val Val Asp 340 Ala Ile 325 Gly Pro Thr Leu Lys Gin 335 Ser Ala Ile Asn Phe Leu 350 Gin Pro Giu Thr Ala Val 345 WO 00/31115 PCT/AU99/01035 54 Lys Lys Asn Tyr Ala Gly Ser Ala Thr Phe Ile Pro Leu Ser Thr Leu 355 360 365 Lys Pro Arg Phe Ile Pro Asp Leu Tyr Leu Glu His Leu Asn Ser Gin 370 375 380 Lys Gly Phe Ile Asn Leu Ala Ser Asn Leu Val Asp Phe Giu Lys Lys 385 390 395 400 Tyr Lys Ile Leu Ala Asp Phe Leu Leu Gly Gly Ile Ile Val Ala Asp 405 410 415 Thr Ile Asp Ser Ala Asn Arg Ile 420 Ala Asn Phe Leu Asn 425 His Lys Asn 430 Met Ile Val 435 Thr Leu Asp Gly Asp 440 Val Ile Arg Thr Ser Gly Ile Ile 445 Ser Gly 450 Gly His Lys Ile Lys 455 Asn Asp Ser Ser Phe Ser 460 Ile Gin Tyr Ile Asp Giu Leu Thr Asn 470 Asn Leu Asn Phe Phe Giu Glu Lys 475 Gin Giu Phe Lys Giu Ser Val Phe 500 Val Lys Ser Asn Glu Phe Glu Gin Leu Ile Thr Arg 485 490 495 Leu Gin Gin Ile Asn Ile Asn Leu Asn Asp Leu Glu 505 510 Gin Lys Phe Ser Asn Ser Giu Asn Giu Leu Ile Glu Ile Lys Ala Gin WO 00/31115 WO 0031115PCT/AU99/01 035 Asn Giu Giy Leu Giu Glu Ser Leu Asn Gin Lys Asp Asp Leu Asn Leu 530 535 540 Ser 545 Leu Asn Arg Leu Giu Leu Giu Thr Leu Lys Giu Lys Ile Giu Leu Giu Asn Val Vai 550 555 560 Asn Gin Cys Lys Ile Leu Lys Thr Giu Lys Lys Gin 565 570 575 Ile Ser Giu Leu Thr Vai Leu Vai Gin Giu Leu Asn 585 590 Leu Asp Gin Lys Lys Asp 61 0 Gin Lys 625 Asn Gin 580 Gin Arg Lys Ile Asn Aia Asp Leu Asn Gin Asn Gin Asn Tyr 595 600 605 Lys Tyr Giu Phe Leu Ile Thr Asn Leu Arg Asn Asn Leu Ser 615 620 Tyr Ser Leu Thr Phe Giu Giy Aia Aia Gin Lys Tyr Giu Leu 630 635 640 Giu Ile Pro Giu Giu Ile Lys Aia 660 Lys Asp Aia Arq Giu Phe Vai Asn Ser Leu Asn Leu 645 650 655 Leu Giy Asn Vai Asn Leu Asp Aia Ile Asn Asp Phe 665 670 Giu Thr Thr Ser Gin Arq Leu Giu Lys Leu Lys Lys Ser Gin Asn Giu WO 00/31115 PCT/AU99/01035 685 Leu Glu Thr Ala Arg Ser Lys Ile Leu Glu Val Ile Ser Asp Leu Asp 690 695 700 Lys Ile Ile Ile Gly Lys Thr Gin Glu Ile Val Asp Leu Val Asn Ser 705 710 715 720 Glu Phe Asn Leu Ile Tyr Phe Ser 740 Ser Ala Gin Pro 755 Val Phe Gin Asn Met Phe Gly Gly Gly Ser Ala Lys 725 730 735 Asp Lys Asn Asp Ile Leu Asn Ser Gly Ile Glu Ile 745 750 Pro Gly Lys Thr Ile Lys Asn Ile Arg Leu Phe Ser 760 765 Gly Gly Glu Lys Ala Ile Ile Ala Ile Ser Leu Leu Phe Ser Ile Ile 770 775 780 Lys 785 Ala Arg Pro Ile Pro Leu Cys Ile Leu Asp Glu Val Glu Ala Ala 790 795 800 Leu Asp Glu Ser Lys Gin Lys Thr 820 Ser Arg Val Asp 835 Asn Val Ile Arg Tyr Val Glu Phe Arg Lys Gin Leu 805 810 815 Gin Phe Leu Ile Ile Thr His Arg His Gly Thr Met 825 830 Gin Leu Leu Gly Ile 840 WO 00/31115 PCT/AU99/01035 <210> <211> <212> <213> <220> <223> 57 31 1698

DNA

Artificial Sequence Description of Artificial Sequence:Clone pAD760 <400> 31 atgaatagaa attcaagcta atttttcagg aaaattgaag gttgaagaaa tttattttac tcaattgaaa tttaagcaaa aaaaataacg attgacgatg aaaataggca caggttgtga cctgaactac tgtaaagtcq tcaattgtag aaaattgagq ccttcaaaag qtagttgttc ctagtagcag aaaaaaccga tccaagatgt aagcaattga ctgatcttga attattttga tatcaaaggc ttaatcttga aaatttctga tagttttggc ataaagtttc ctaaacatga tatcctcaaa agtcaaaatt caattcgcag gagcaactqg tgattaaata ttatttgcgt ctgattttca atctaacacg aaaatcaaaa ggcaaaaaat atttgtaata acaattgagt tgatcttaca aaaagaaact aagttttgaa aattgtccga cactgttact tqcctttatg agttgtaatt atcaqttcaa tcttqaaatc aaatgaagat ctcaagaqtt tgatgataat aaaagagttc acatagttta ttgtcaagtg attagttcaa agcgaattaa acaaagaaaa tttaaagttt gatgaggaaa qatcctgaga cattgacttt aataatgttt aataaaattg ccaagccatt gaaaatgtat cttgtcaaga acttcaattg gccgaagctq cttgcaattt attgtcaaat aaaataggtc gccattggaa caaattatcc aagatagtaa atctggaagc ttqacccaga ttaatacaaa aagttaacga ttcaggttga ttatggcaat ataacaaata ctgctggtta atgcaagtgg caaaaaatac aaaaaataat cacggattcc ataatatttc ctcaagaact ttattgttaa ataaattacg aaaacggttc cgtattcaag aaaattaata agttattgat tgcgcaaata cgggattgtt tcttgtttct tgatattttt tatgcacgct tttatcgctt tattttcgaa acaaaattta aaaacagtcg cgatgcaatc agqggaaaga tgaaatcggc tcaaggtgaa tgcgatgtcg tcgttttatc aaatgttaaa cgcgctaaaa 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 WO 00/31115 WO 0031115PCT/AU99/O1 035 gatcaaaatt cttaacaatg tatggtcaag gagttagaaa gtaagaagtc tcgcgtaatt ggctttgaaa gatgcaqatt gataaaattc tgagcaaaaa ttaaaattga agtatattca gtaataataa aaccttatgg accaagaatt ttaacaaaag acgaqaaaga cactttttga accaagaaga atgaagct atgaaatgga tcqccagcag ttttgactta gattgaaaac tccaaaaaac ccagaatatc ttctaattct ttcggctcta ggaaaaaaaa aatattaaaq ggtagaattt attttacaqc gagtttatac aagaataatc cgcgaaaata ttccaaaata aatgaggcaa caagaattat acattcaaga atcaaaatca aattcgaatc caaaaaatga taqcaaaaqc tttcaaaaqa ttagtcaaag ttagtgaaaa tattacaaga actaaacagt tcqgaattca tgatattcgc gcaaaaacag aqcaactaaa ttttgattat atcttttttt cgagttaatc gaaggaaaaa 1200 1260 1320 1380 1440 1500 1560 1620 1680 1698 <210> <211> <212> <213> <220> <223> 32 566

PRT

Artificial Sequence Description of Artificial Sequence:Deduced protein sequence from clone pAD76O <400> 32 Met Asn Arg Lys Lys Thr Glu Lys Ser Lys Ile Ser Ser Lys Asp Ser 1 5 10 Lys Lys Leu Leu. Asn Leu Ile Ile Gin Ala Ile Glu Ala Val Ile Asp 40 Gin Asp Val Ala Lys Asn Ser Glu 25 Ile Phe Gin Giu. Ala Ile Glu Phe Val Ile Thr Lys Lys Ile Asp Pro Asp Ala Gin Ile Lys Ile Glu Ala WO 00/31115 WO 0031115PCT/AU99/01 035 Asp Leu Glu Gin Leu Val Glu Glu Asn Tyr Ser Phe Lys Val Phe Asn Thr Asn Gly Ile Val 70 75 Phe Asp Asp Leu Thr Asp Giu Giu Lys Val Asn 90 Ile Leu Leu Ser Lys Ala Lys Glu Thr Asp Pro 105 110 Asp Leu Val Ser Phe 100 Giu Ile Gin 115 Val Asp Asp Ile Phe Ser Ile Giu Ile 120 Asn Leu Glu Ser 125 Phe Lys Gin Lys Phe Giu 130 His Trp Leu Phe Met Ala Ile Met His 135 Ala 140 Ser Glu Ile Val Arg 150 Asn Asn Val Tyr Asn 155 Lys Tyr Leu Ser Lys Asn Asn Val Val 165 Leu Ala Thr Val Asn Lys Ile Ala Ala Gly 175 Tyr Ile Phe Giu Ile Asp Asp Asp Lys 180 185 His Tyr Ala Ser Gly Gin Asn Leu Lys 195 200 Val Ser Ala Phe Ile Gly Thr Lys 205 Met Pro Ser 190 His Giu Val Val Ile Glu Asn Vai Ser Lys Asn Thr Lys Gin Ser Gin Vai Val Ile T/AU99/01 035 210 215 220 Ser Ser Lys Ser Val Gin Leu Val Lys Lys Lys Ile Ile Asp Ala Ile 225 230 235 240 Pro Glu Leu Gin Ser Lys Phe Leu Giu Ile Thr Ser Ile Ala Ar Ile 245 250 255 Pro Gly Giu Arg Cys Lys Val Ala Ile Arg Arg Asn Glu Asp Ala Glu 260 265 270 Ala Asp Asn Ile Ser Glu Ile Gly Ser Ile Val Gly Ala Thr Gly Ser 275 280 285 Arq Val Leu Ala Ile Ser Gin Giu Leu Gin Gly Giu Lys Ile Glu Val 290 295 300 Ile Lys Tyr Asp Asp Asn Ile Val Lys Phe Ile Val Asn Ala Met Ser 305 310 315 320 Pro Ser Lys Val Ile Cys Val Lys Giu Phe Lys Ile Gly His Lys Leu 325 330 335 Arg Arg Phe Ile Val Val Val Pro Asp Phe Gin His Ser Leu Ala Ile 340 345 350 Gly Lys Asn Gly Ser Asn Val Lys Leu Val Ala Asp Leu Thr Arq Cys 355 360 365 Gin Val Gin Ile Ile Pro Tyr Ser Ser Ala Leu Lys Asp Gin Asn Phe 370 375 380 WO 00/31115 PCT/AU99/01035 61 Gly Asn Ile Lys Asp Ile Gin Giu Leu Asn Ser 390 395 400 Ile His Arg Gin Gin Giy Ar Ile Tyr Gin Asn 410 415 Lys Ilie Giu Trp Asn 385 Leu Asn Asn Giu Tvr His Arg Asn Gin Gin Phe 435 Tyr Giy Gin Giy Asn Asn Asn Phe Asp Leu Ile Leu 425 430 Giu Leu Giu Lys Pro Tyr Giy Ile 445 Giu Ser Asp Ile Giu Asn 450 Gin Giu 465 Giu Phe Ile Pro Lys Asn Giu Gin Lys Gin Vai Arg Ser His 455 460 Lys Asn Asn Leu Aia Lys Ala Ala Thr Lys 475 480 Phe Pro Lys Ser Arg Asn Phe Asn 485 Lys Ser Gin Asn Ile Arq Giu Asn Ile Ser Lys 490 495 Asp Phe Asp Asn Ile Ser 515 Aia Leu Asn 530 Giy Phe Giu Asn Gin Arg Ser Phe Phe 520 Giu Aia Ile Ser Giu 535 Giu Lys Asp Ser Asn Ser Phe Gin 505 510 Asp Ala Asp Ser Leu Phe Asp Ser 525 Asn Giu Leu Ile Asp Lys Ile His 540 WO 00/31115 PCT/AU99/01035 62 Gin Giu Glu Glu Lys Lys Gin Glu Leu Leu Leu Gin Glu Lys Glu Lys 545 550 555 560 Trp Ala Lys Asn Giu Ala 565 <210> 33 <211> 1860 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence:Clone pAD774 <400> 33 gaattctacc gaagacaaaa caaattaatg aaaataactg aatqaaagtc tcaaaattag attcgagatt ctagaaaatg ccaatccact ggaaacggtc aattcaagtt tttgttgact acctttaaaa gCtqttttac tttqatcacc ttccggctga cctcttttaa aqqcacttga caaaagcata aaggatttaa tttcatctat atgtttttag cttgttataa attttagcac attcaacaga ccgcccataa attcaattct ctgccggatt taatgcaaat aattattgtg tggatattcc tccttatgcc tttaattgac caaaaatccg tagtaaaaaa ttatttaaat tttcagccaa ttataattqg tccaatqgat aaataagatc tgaggatgta tccqccactt cgqgtacatt atgtcaaaaa tataatttat tttagtattg attttttccg caaattgatq acagaaaata agcttaaaat aaaaacgcta ggttatgacc ccatatctgc ggaattgttc gctcacgggc gatacacgaa ttgcaattta ataatgatqt taattgagca cgccttttga aaaaaactgg ctcaaattta aaggtacttt ttaatttttt gcatcctcca caattggtta gaatttttqa ttqatgttga atttttaccg aaccaatggc taaagatccg ttggttttgc tcacgattta ttgaaaaaaa aaaaaatcct tcagctgcac tatcqgagcg acaattatta caaaggggat cttttcgata gtttaaaaaa ttttagtcac ggatgaagca atttaggcta 120 180 240 300 360 420 480 540 600 660 720 780 840 WO 00/31115 WO 0031115PCT/AU99/01 035 attttggatt ttagcatctt aatccaaata cgacttgaaa caaacagcaa agtaaaaaat caaaatttgc cttgcctatc acgcgaattg tttgtccaaq gatttttctg ttttcgtttt aatggattga cttgccgaaa aaaatcaaaa aaaattctgc aatatgattt ccttaatttt t ttt agat aa ttcttttata aagggaaaac ttatcgggag tcgctgctta catattcaaa ttacttgtta gaaaaagaga gaaaaatttc ggggcagtta tagcgggcaa attttgaaat ttaatcaatt aacagctaaa taaaaataaa taaaaatttt ttttactaaa aaaagcaatt tggtgatttt aggaaatagt cggaaatccg tgtttcttca aaaaaaatac taatggcccg atttcttgaa acttagtgct tcaaaattgc atatcttgat tcttaaaaac tcgccaaaat atttgaaact gtgattaaaq aatattcttt tattataaag acagttattg tcaaatccta tttaactttg cqtgataaag attccgggqa gacctttttg actttagccg atctaccqgc gggactgaat tatcctaatt tttcattctg aatgaaatca tctccatttt gttgataaca ttattcataa ttagctcaaa ttgatggatt ccagtgaact gtgacctacc gatatttaaa gtttaatttt acattgcaaa caaacgatat ataaaattct aagccttaat ttgctttttc taaacataaa ataaaaccac aagagaaaat ttcggcttga ataaaggatt tttttcacat gattaaagtt tcgctttaat aaaaaaagtt aagcagaaat cgatacaatc atcaaaaact ttttgacttc cagcctaaat tagtgcgaca gatggtcaat aagaatttgt aagaccgcct agatttcaca ctttgatttt tacaaaccaa aattatcttt ctttcttatg attcctaatt 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 <210> <211> <212> <213> <220> <223> 34 620

PRT

Artificial Sequence Description of Artificial Sequence:Delucei protein sequence from clone pAD774 <400> 34 Glu Phe Tyr Leu Trp Ser Pro Asn Ala Asn Arg Val His Phe Ala Ile 1 5 10 WO 00/31115 WO 0031115PCT/AU99/01035 Tyr Lys Asp Pro Giu Asp Lys Ile Lys Asn Asn Asp Val Trp Phe Cys 40 Tyr Ser Tyr Asn Leu Leu Ile Giu 55 Pro Ala Giu Ile Ile 25 Val Met Ser Phe Asn Gly Gin Ile Asn Ala His His Asp Leu Ser Lys Ile Thr Giu Ala Leu Asp Pro Tyr Ala Phe Ser Ile Ala Pro Phe 70 75 Asp Trp Lys Lys Asn Glu Ser Pro Lys Ala Tyr Leu Ile Asp Ile Phe Ser Giu Lys Thr 90 Gly Lys Asn Asp Ala Gin Lys Lys Thr 130 Pro Ser Lys Leu Giu Giy Phe Asn Lys Asn Pro Gin Ile 100 105 110 Ile Tyr Gin Leu His Ile Arq Asp Phe Ser Ser Ile Ser 120 125 Glu Asn Lys Gly Thr Phe Ile Giy Ala Leu Giu Asn Asp 135 140 Val Phe Ser Tyr Leu Asn Ser Leu Lys Phe Asn Phe Leu Gin Leu Leu 145 150 155 160 Pro Ile His Ser Cys Tyr Asn Phe Ser Gin Lys Asn Ala Ser Ile Leu 165 170 175 WO 00/31115 PCT/AU99/01035 Asp Gly Asn Gly His Phe Ser Thr Tyr Asn 180 185 His Lys Gly Asp Pro Ile Giy Tyr Phe Ser Ile Asn Ser Ser Tyr Ser 195 200 205 Met Asp Pro Tyr Leu Arg Ile Phe Glu Phe Lys Lys Phe 210 215 220 Trp Gly Tyr 190 Thr Asp Pro Val Asp Ser Ala 225 His Lys Asn Lys Ile Gly Ile Val Leu Asp Val Asp Phe Ser His 230 235 240 Thr Phe Lys Asn Ile Leu Giu Asp Val Ala His Gly His Phe Tyr 250 255 Gly Phe Pro Pro Leu Asp Thr 270 Arg Asp Giu Arg Lys Pro 275 Ala 260 Ala Vai Leu Pro Met Ala Phe Arq Leu Ile Leu Asp Ser Leu Ile Phe Phe 280 285 Gly Phe Arg Phe Asn Leu Ala Ser Phe 300 Thr Lys 290 Tyr Tyr Lys Val Leu 305 Asp Lys Lys Ala Ile 310 Thr Val Ile Ala Ser Giu Leu Lys Lys Val 315 320 Tyr Gly Asp Phe Ser Asn Pro Ser Asp Leu 330 335 Asn Pro Asn Ile Leu Leu 325 WO 00/31115 PCT/AU99/01035 Pro Ser Arg Asn Arg Leu Glu Lys 340 Phe Gly Tyr Leu Asn Asp Thr Ile 355 360 Gly Lys Thr Gly Asn 345 Ser Phe Asn 350 Gly Ser Gly Gin Thr Ala Ile Asn Pro 370 Arg Asp Lys Gly Ile Leu Ser Lys Thr 380 Ser Lys Lys Phe Ala 385 Ala Tyr Val Ser Ser 390 Ile Pro Gly Asn Ala Asn Phe Asp Gin Asn Leu Pro Tyr Ser 405 Lys Lys Lys Asp Leu Phe Ala Asn Asp 415 Ile Ser Leu Ala Asp Lys 435 Asn Leu Ala Tyr Leu Thr 420 425 Ile Leu Ser Ala Thr Thr 440 Cys Tyr Asn Gly Pro Thr Leu 430 Arg Glu Phe Arg Ile Gly Leu Glu 450 Ile Tyr Arg Gin Ala Leu Met Met Val 455 Asn Phe Val Gin Gly 460 Phe Ser Arg Ile Cys 480 Lys 465 Ile Ser Leu Ser Ala Gly Thr Glu Phe 470 Ala 475 Asp Phe Ser Gly Gly Ser Tyr Gin Asn Cys 485 490 Tyr Pro Asn Leu Asn Ile 495 Lys Arg Pro Pro Phe Ser Phe Leu Ala Gly Lys Tyr Leu Asp Phe His WO 00/31115 WO 0031115PCT/AU99/01 035 Ser Asp Lys Thr Thr Asp Phe Thr Asn Gly Leu Asn 515 520 Lys Asn Asn Giu Ile Lys Giu Lys Ile Phe Asp Phe 530 535 540 Asn Gin Phe Arq Gin Asn Ser Pro Phe Phe Arq Leu 545 550 555 Glu Ile Leu Leu Ala Giu Ile Asp Thr Asn Lys Ile Lys Lys Gin Leu Lys Phe Giu Thr Val Asp Asn Asn Lys Gly 565 570 575 Leu Ile Ile Phe Lys Ile Leu Leu Lys Ile Lys Trp Leu. Lys Leu Phe 580 585 590 Ile Ile Phe His Ile Phe Leu Met Asn Met Ile Leu Lys Ile Leu Ile 595 600 605 Phe Phe Leu Ala Gin Arg Leu. Lys Leu Phe Leu Ile 610 615 620 <210> <211> 897 <212> DNA <213> Artificial Sequence <220> WO0/31115 68 <223> Description of Artificial Sequence:Clone pAD784 PCT/AU99/01 035 <400> gcttttttgt caggtttttt aatcggcatt ccaaatgccg gaaatcaaaa ccaaaaatca aagcttttaq gcaatgccgc gattttggtg cagattagtg agaaaaaaag atcaccgaac gatatccatg gtaataactg gttgttttag ttgttgatgg tacctaccaa gatctgaata attttgttaa atatcgaaaa aaccgggaat gagccgaaaa attgaagggc cgctttttaa aaaaaaaatt ttgctcctgg aaggctatgg aattgccagt ttgaacccgg taactgaaag tcggtatatt gcaaatcttg tttaactatt gttacaagcc agctgttgaa qacgqagaaa agaatccttt gagtgaaacc cggttattgc agaaattttg ggttaaagcc aaaatatttt tgttagttca aatttatatc tggttttcgt gaaaaaqctg aagatttttt gaccaatttg caactttttc atctcactcg agtatcgatg gattcaataa gaaattttag gctgatatta gaaatagtaa agcgaaattg attcactcaa actagccaga cctggacttg accttgtcac aaaaagatqc caagaaaacc ataaaataag gaattataaa cggcttataa tcaacctaaa ttgcaactgg ataatgatct caagaacttc aaaaagctgc accttgcttg ctggccacgg caattttaga ga ggcgcaag gaaaaggtga taaaaattgt cgtatcaaaa aagttgattt aacagaagaa taaaatattt ttatcaaatg ttctaattcg tttaaaaatt ttatcttgga tgaaattggt ccggaatttt ggttggtatt gcccggaatg aattgaggat aagaat t 120 180 240 300 360 420 480 540 600 660 720 780 840 897 <210> <211> <212> <213> <220> <223> 36 299

PRT

Artificial Sequence Description of Artificial Sequence: Deduced protein sequence from clone pAD784 <400> 36 Ala Phe Leu Phe Val Asp Gly Arg Tyr Ile Glu Lys Ala Glu Lys Asp 1 5 10 wnAA 1114 ID T/AU99/01035 69 Ala Lys Asn Cys Gin Val Phe Leu Pro Thr Lys Gin Ile Leu Lys Ile 25 Phe Ser Arg Lys Pro Val Ser Lys Asn Arg His Trp Ser Giu Tyr Leu 40 Thr Ile Asp Gin Phe Asp Lys Ile Arg Ser Trp Phe Pro Asn Ala Asp 55 Phe Val Lys Leu Gin Ala Gin Leu Phe Arg Ile Ile Lys Thr Giu Giu 70 75 Giu Ile Lys Asn Ile Giu Lys Ala Val Giu Ile Ser Leu Ala Ala Tyr 90 Asn Lys Ile Phe Pro Lys Ile Lys 100 Pro Gly Met Thr Giu Lys Ser Ile 105 110 Asp Val Asn 115 Leu Asn Tyr Gin Lys Leu Leu Gly Ala Giu Lys Giu 125 Ala Met Pro His Ser Phe 130 Asp Ser Ile Ile Ala Thr Gly Ser Asn 135 T rp 145 Arg Ala Ser Giu Thr 150 Giu Ile Leu Asp Asn 155 Asn Gly Tyr Cys Ala 170 Asp Leu Leu Lys Ile 160 Asp Ilie Thr Arq Thr 175 Asp Phe Gly Ala Leu Phe 165 WO 00/31115 WO 0031115PCT/AU99/01 035 Ser Tyr Leu Gly Gin Ile Ser Giu 180 Lys Lys LeU Glu Ile Leu Giu Ile 185 190 Val Lys Lys Ala Ala Giu Ile Gly 195 200 Lys Ala Ser Glu Ile Asp Leu Ala 210 215 Gly Tyr Gly Lys Tyr Phe Ile His 225 230 Arg Lys Lys Val Cys Arq Asn Ala Pro Gly Val 205 Ile Thr Giu Gin Gly Val Gly Ile 240 Ser Thr Gly His 235 Asp Ile His Glu Leu Pro Val Val Ser Ser 245 250 Thr Ser Gin Thr Ile Leu 255 Glu Pro Giy Met Vai Ile Thr Val Giu 260 265 Pro Giy Ile Tyr Ile Pro Giy 270 Leu Gly Gly Ala Ar Ile Giu Asp 275 280 Phe Arg Thr Leu Ser Arg Lys Gly 290 295 Val Val Leu Val Thr Giu Ser Giy 285 Giu Ar Ile <210> 37 <211> 633 <212> DNA <213> Artificial Sequence WO 00/31115 71 <220> <223> Description of Artificial Sequence:Clone pAD789 PCT/AU99/01 035 <400> 37 gatcttgtat tctttaatta ttaaatttag aagtccacat ataaataaca tatgtataca gaaccgaaaa gatttagaca aatccccttg tccaacttag ttaacattaa ttaaagtaga gtattaaaaa aagttcaaaa tttcaaqgat aatacgatat acaattcaga caaaagatga agaaaattga cacgctcact accaagaaag ttttcattac aaattctgaa tttaaactat aggtaaaaaa tttattaggc aaaaaaaata cttaattaat gattaggccg ttctgatgtt ttattctcca ttatgttaaa ccctcacttt aatcaattac aaattggaag tatttactaa attgagaagq aatcctgatt gcatcaactc ttattaqaaa tgattatttt aaagaaattt atcatatctg gat aagatttaga atagagtttt aagqagctaa aatttgaagq ctataaataa taqaaaatat aqgtaaattt tccactggqc taataattga aacttattgc tggaactttt atttaataat cqggtctgga tcaaatttta ccatattaat ttcgcttttg tgaaaacctt aaatccaaaa cgaaggtctt ggataaaaat 120 180 240 300 360 420 480 540 600 633 <210> 38 <211> 211 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence:Deduced protein sequence from clone pAD789 <400> 38 Asp Leu Val Phe Lys Val Glu Asn Ser Glu Asn Gin Leu Gin Asp Leu 1 5 10 Asp Gly Thr Phe Ser Leu Ile Ser Ile Lys Asn Leu Asn Tyr Lys Leu WO 00/31115 WO 0031115PCT/AU99/01035 Giu Asp Arg Vai Leu Phe Asn Asn Leu Asn Leu Giu Val Gin Lys Gly 40 Lys Lys Tyr Leu Leu Lys Gly Ala Asn Gly Ser 55 Ser Arg Ile Leu Leu Gly Ile Giu Lys Giu Phe 70 75 Ile Asn Asn Lys Tyr Asp Ile Lys Lys Ile Asn 90 Gly Lys Ser Thr Phe Glu Gly Gin Ile Leu Pro Asp Ser Ile Asn Asn Hi-s Ile Asn Tyr Val Tyr Asn Asn Ser Asp Leu Ile 100 105 Asn Ala Ser 110 Asp Giu Ile Thr Leu Glu Asn Ile Ser Leu Leu Giu Pro Lys Thr Lys 115 120 125 Arg Pro 130 Leu Leu Giu Lys Vai Asn Phe Giu Asn Leu 135 140 Trp Leu Phe Phe His Trp 155 Asp Leu Asp Lys Lys 145 Ile Asp Ser Asp Ala Asn Pro Asn Pro Leu Ala Arq 165 Asp Giu Giy Leu Ser 180 Ser Leu Tyr Ser Pro Lys Glu Ile Leu Ile Ile 170 175 Asn Leu Asp Gin Giu Ser Tyr Vai Lys Ile Ile 185 190 WO 00/31115 PCT/AU99/01035 73 Ser Glu Leu Ile Ala Asp Lys Asn Leu Thr Leu Ile Phe Ile Thr Pro 195 200 205 His Phe Asp 210 <210> 39 <211> 1791 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence:Clones pAD9O8, pAD98l, pAD1013, and pAD1O49 <400> 39 cttgaaataa gggaaaaqat ttttgggaga ccttgtagta aagtataata aatttaaaat tcacgactgt qcaaccttat ctaaccggaa gacaaattaa gagcatgatt qgtgaaaacg tcaaatatgg atgactattt catcaagcga gttgcaaagg tctttcaatt taactgaatt catttttagc caggaqqcga ttctttatgt t tgcgacact ttqagacaat gtggtttttt atcaaaagaa tcgaccaatt agatttacga agcccggctt atcccaactt tqacggaaaa aaatqttggt atcccaacga tcttgatgaa gaaaaaaatg tttagctgct ggttgctgca tctctaaatt actgactaat ttctgaaagc gaaggaattc ctttgattta aacaagtatg tcaattaaag atttctactt cttgaatatt attagqcttg ccttcaattg gttgaaattg gattatattg ggatcaatta ttaataaaat agaaaatgat cgcttgcCgt atttaataac taattctcga taactttaaa caagccaggt gcctacatca gtaatagttt ttgatatcgg aggatattga tcagagaaaa gtcagaattt tttcattgaa tt ttt tt aga tgaaattaaa tagatcaacg tgqatatcaa aaaagataat aattgtagtc gccaaaagct aaatgaacca WO 00131115 WO 0031115PCT/AU99/O1 035 aaatcactta cgggctggca attaatgtca aagtcgacat aatattcgca ttgcaatcaa gtttttgatg tcaaagtcaa gggcaaatta caaggaaaaa gatattcttg atcgcaaaat gcaataactt aaacctaaag gatgttgcta gtaatcgagc aacggcgggg gttagtgaat ccggccaatt atggcaaatt acattcctct tagttaatca ttgggaaaaa tcaaagtcaa atatccgtga aattttcctt aaattgaaat gatacaagcc atttaacagt tacaaaccct tatcaggagg gaaaatcact atttaattaa ataatttaga ataacggggg catatactgg tttaactgga tataattatt aqgcaaattt aattatcqta tgtgaggaaa tcggacgaac ggtttttgcc taatctgcaa gtattttatg ggatgtccta ttcagaagca agctgatgtt agaagctcaa ttttgtactt aqtgctaaat ggtaattaaa ccaaatagtt cgcttattta aaattggaaa gaaaaagctg qttgttgtca aatgcgattg ttaaagggct cctagatcaa aatactgagc actgggcggt cctgatattt caaattcgtt cttgaatttt ggtcttggtt cgaattaaat gacgagccaa cgaatagtcg gttgctqact gcaaaaggaa aaaacaattt ttccaqtacc ctqaaaataa ctggtgtttC ccaaaaatct ttttaatatt atcccgcaac aggcaagagc gtgataaatg atgttttatg tttatgqaaa tccataattg atataaaact tagccacttt caactggact ataacggtga at attattga caccagaagc aaaaaaacga tttaaaaaaa tggatctqqa tggaacaact gataagttga ttatacctct gcttggtttt ccaaggagac tgatcactgc aacaatcgcg gcctaaaata tqgccaatca tttacaaaaa ccataattat tagcataatt tttaggccca tgtagcaaaa 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1791 taaatataaa a <210> <211> <212> <213> <220> <223> 597

PRT

Artificial Sequence Description of Artificial Sequence: Deduced protein sequence from clones pAD9O8, pAD981, pAD1O13, and pAD1O49 <400> Leu Glu Ile Ile Lys Tyr Gly Ser Lys Glu Ser Leu Asn Tyr Trp Leu WO 00/31115 WO 0031115PCT/AU99/01 035 Ile Ser Giu Ser Gly Lys Arq Tyr Ile Leu Asp Lys Ile Gin Arq Lys 40 Asp Tyr Phe Arg Pro Ile Giu Gly 25 Phe Trp Giu Thr Ser Ser Giu Asp Leu Arg Leu Trp Phe Lys Lys 55 Cys Lys Giy Ala Arg Leu Asn 70 Lys Tyr Asn Ile Phe Gin Leu Met Met Ser Giu Phe Pro Cys Ser Ser Lys Tyr Ala Aia Vai Phe Ile Giu Ser Gin Ser Ile Lys Asp Leu Ile Thr Phe Phe Arq Asn Leu Lys Leu 100 Thr Leu Ile Leu Asp Giu Ilie Lys 115 120 Vai Giy Leu Giu Tyr Leu Thr Leu 130 135 Giu Phe Asp Gly Lys Ile Ser 110 Leu Aia Asn Ser Arg Leu Ser Arg Asn Arg Ser Leu Aia Ser 155 Thr 140 Ala Thr Leu Ser Gi y 145 Gly Giu Ser Gin Arg Ile 150 Gin Vai Giy Tyr Gin 160 Leu Thr Giy Ile Leu Tyr Vai Leu Asp Giu Pro Ser Ile Giy Leu His WO 00/31115 WO 0031115PCT/AU99/01 035 165 Gin Lys Asp Asn Asp Lys Leu Ile Ala 180 185 Ile Gly Asn Ser Leu Ile Val Val Giu 195 200 Thr Leu Lys Lys Met Val Giu 190 Thr Ile Leu His Asp Phe Giu 205 Ala Ala Asp Tyr Ile Val Asp Ile Gly Pro Lys Ala 210 215 220 Gly Phe Leu Val Ala Ala Gly Ser Ile Lys Asp Ile 225 230 235 Gly Giu Asn Gly Giu Asn Giu Lys Ser Leu Thr Gly Gin Phe Leu Thr Gly Lys Leu Giu Ile Pro Val 245 250 255 Pro Lys Lys Arg Arg Ala Giy Asn Gly Lys Phe Ile Ile Ile Giu Lys 260 265 270 Ala Ala Giu Asn Asn Leu Lys Lys Ile Asn Val Asn Ile 275 280 285 Pro Leu Gly Ser Thr Leu Lys Phe 290 Val Val Val Thr Gly Val Ser Gly Ser Gly Lys 295 300 Val 305 Asn Gin Ile Ile Val Asn Ala Ile Ala Lys Asn Leu Gly Thr Thr 310 315 320 Asn Ile Arg Ile Gly Lys Asn Val. Arg Lys Leu Lys Gly Phe Leu Ile 325 330 335 WO 00/31115 WO 0031115PCT/AU99/01 035 Leu Ile Ser Trp Leu Gin Ser Ile 340 Lys Val Asn Arg Thr Asn Pro Arg 345 350 Ser Asn Pro Ala Thr Tyr Thr 355 Phe Ala Asn Thr Giu Gin Ala 370 375 Se r 360 Val Phe Asp Asp Ile Arg Giu Vai 365 Ser Lys Ser Lys Arg Ala Leu Giy Phe Ser Phe Asn Leu 385 Gly Gin Ile Lys Ile 405 Cys Asp His Cys Gin 420 Gin 390 Thr Gly Arg Cys Lys Cys Gin Giy Asp 400 Giu Met Tyr Phe Pro Asp Ile Tyr Vai Leu 415 Gly Lys Arg Lys Pro Asp Vai Leu Gin Ile 430 Thr Vai Ser Arg Phe Tyr Gly Lys Thr Ile 435 Giu Ala Leu Giu Phe Phe His 450 455 Al a 440 Asp Ile Leu Asp Asn Trp, Pro Lys Ile Ile Aia Lys Leu 460 Gin 465 Thr Leu Ala Asp Val 470 Gly Leu Giy Tyr Ile 475 Gly Giu Aia Gin Arg 490 Lys Leu Giy Gin Ser 480 Ile Lys Leu Ala Thr 495 Ala Ile Thr Leu Ser Giy 485 WO 00/31115 WO 0031115PCT/AU99/01 035 Phe Leu Gin Lys Lys Pro Lys Gly 500 Pro Thr Thr Gi Leu His Asn Tyr 515 520 Lys Ser Leu Phe Val Leu Asp Glu 505 510 Asp Val Ala Asn Leu Ile Lys Val 525 Leu Asn Arg Ile Val Asp 530 Asn Leu Giu Val Ile Lys 545 550 Asn Gly Giy Asp Asn Gly 565 Gly Asp Ser Ile Ile Vai Ile Giu His 540 Val Ala Asp Tyr Gly Gin Ile Val 570 Ile Asp Leu Giy Ala Lys Giy Thr Pro Giu 575 Giy Ala Tyr Leu Lys Thr 590 Ala Vai Ala'Lys Val Ser Giu Ser Tyr Thr 580 585 Ile Leu Asn Ile Lys 595 <210> 41 <211> 900 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence:Clone pAD913 WO 00/31115 WO 0031115PCT/AU99/01 035 <400> 41 atgaaaatta aaagcccttq caaactggtt caaggcgaaa caaaaaccgc ggcgtcgttt atatttgggg ataattgaat ggtggccaga ttttttgatg cttgatactt aagcaaaaac ataaagtaac caggaaaaac ttgagtatct gtttttttag ttcagtttgc ctgtttcaat tagttqgtct aacgccgagt aacccacggc tatataaaaa catcgtaaaa tactqaaata aacttttatt ctattttgat aaaaaaacta tgaatatcag gggaactcca tgatcaaagt tgcaattgcc cggacttgat gggcaagaca ttgtattttt aaataataaa aaaattaatc tgaaattaat atttatgatc aatcagggcg cagcatatga tcaaaaaatc aaatttatta ctttttgaqc aaaaatgagq tttttacaaa qgaattttag ccccaaggaa atcattctgg tttaaagatg tttttaattg aaaattggtt atgctaattc aaaaattacc agtttattgc atgcactttt aagaaaaaaa atgaaattcg aaacaattga caaaaaaaat aatcaccttt caatqgatcc cgctaaaaat caactcatga gtagaattat aaaataagat atccaatttc aaaaggaaac atcagaatta aataattggc gctaccagat attagttgtt tcggcgtgtg aaaaqacatc tgccgcagaa tgaactttca tgatattatt gcttgaaatt tcttgatagt ttatgatggt gttaccaact taatgttaga aaatgcagat 120 180 240 300 360 420 480 540 600 660 720 780 840 900 gttttagaat gaacaaaacg gatacttatt aatttactca tcagtatctg caattttagc attttcgcga agttaatcag <210> 42 <211> 300 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence:Deduced protein sequence from clone pAD913 <400> 42 Met Lys Ile Lys Ala Lys Thr Ile Val Lys Ile Tyr Asp Gln Lys Leu 1 5 10 WO 00/31115 WO 0031115PCT/AU99/O1 035 Pro Ser Giu Leu Lys Ala Leu Asp Lys Vai Thr Thr Giu Ile Asn Gin 25 Gly Giu Phe Ile Ala Ile Ile Gly Gin Thr Gly Ser Gly Lys Thr Thr 40 Phe Ile Gin His Met Asn Ala Leu Leu Leu Pro Asp Gin Gly Giu Ile 55 Giu Tyr Leu Tyr Phe Asp Ser 70 Lys Asn Gin Giu Lys Lys Leu Val Vai 75 Lys Phe Ile Asn Glu Ile Gin Lys Pro Arg Phe Phe Arg Lys Lys Leu 90 Arg Arg Arg Giu Gin Thr 115 Val Giy Val Val Phe Gin 100 105 Ile Giu Lys Asp Ile Ile 120 Phe Ala Giu Tyr Gin Leu Phe 110 Ser Met Gly Phe Gly Ala Thr Pro 130 Lys Asn Giu Ala Lys Lys Ile Ala Ala Glu 135 140 Ile Ile Giu Leu Val 145 Gly Leu Asp Gin Ser Phe Leu Gin Lys Ser 150 155 Pro Phe Giu Leu Giy Gly Gin Lys Arg Arg Val Ala Ile Ala 165 170 Giy Ile Leu Ala Met Asp 175 Pro Asp Ile Ile Phe Phe Asp Giu Pro Thr Ala Gly Leu Asp Pro Gin WO 00/31115 WO 0031115PCT/AU99/01 035 Gly Thr Leu 195 Lys Met Leu Glu Leu Asp Thr Leu Tyr Lys Lys Gly 205 Val Leu Glu Trp Lys Thr 210 Ile Ile Leu Ala Thr 215 His Asp Leu Asp Lys Arg Cys Ile Phe Lys Asp Gly Ile Ile Tyr Asp Giy 240 Asp Thr Tyr Ser Leu Ala Asn Asn Phe Leu Ile Giu Asn Lys 255 Met Leu Pro Gly Tyr Pro 275 Ile Asn Met 290 Asn Leu Leu Asn Ile Ser Asn Val Arq 280 Leu Ile Gin Lys Giu 295 Phe Arg Glu Lys Leu Ile Lys Ile 265 270 Ser Val Ser Giu Leu Ile Ser Giu 285 Thr Asn Ala Asp 300 <210> <211> <212> <213> <220> 43 489

DNA

Artificial Sequence WO 00/31115 82 <223> Description of Artificial Sequence:Deduced protein sequence from clone pAD920 PCT/AU99/01 035 <400> 43 ttaaaatccc caagaaaaag atttttcctg ttaqatcaaa attgggagtt acatcaattg gcaacaaaac gatttqtcaa ttatcaagt gtttttatca ataaaaaaac aaagtttgct atgaaaaaat tcgcttttca gaaaacaagc taqaaacaat aaaatttgat aaaagtaaat tgaqcccgaa tgagattgat tcaagaactt aggaaacaat ttttgcaaaa tgaacctggt tttagccgaa tcgcaqatag aaaattaatt gattatgcct gatttttcaa ataaaaacgc aacagtttag gccttttttg attttcaggg acgttaaaaa tttatactct tttatgtcga aagcaatgaa tcgttttacc aaacagtaga ataataaaat tagttttgag aaacacagat gaaaaaagta tagtgcaaat attaaggagg cccttctatt tttttcccaq caccgaactt acaaatcaaa 120 180 240 300 360 420 480 489 <210> <211> <212> <213> <220> <223> 44 163

PRT

Artificial Sequence Description of Artificial Sequence:Deciuced protein sequence from clone pAD92O <400> 44 Leu Lys Ser Arg Phe Tyr Gln Lys Val 1 5 Lys Asn Thr Asp Gin Glu Lys Asp Lys 25 Asn Ser Gin Ile Asp Val Lys 10 Lys Thr Glu Pro Glu Lys Ile Asn Phe Tyr Thr Leu Lys Lys Val Ile Phe Pro Giu Ser Leu Leu Glu WO 00/31115 WO 0031115PCT/AU99/01 035 Ile Asp Giu Lys .31)4u Asp Tyr Ala Phe Tyr Val Asp Ser Ala Asn Leu Asp Gin Asn 55 Ile Gin Giu Leu Asp Phe Ser Lys Ala Met Lys Leu Arq Arg 70 75 Ile Gly Ser Phe Ala Phe Gin Giy Asn Asn 90 Ilie Lys Thr Leu Val Leu Pro Pro Ser Ile i00 Leu Giu Thr Vai 115 Thr Ser Ile Giy Lys Gin 105 Ala Phe Ala Lys Asn Ser 110 Thr Ile Giu Asp Phe Ser Ala Thr Lys Leu Pro Gly Ala Phe Phe Asp Asn 130 135 Asn Leu Ile Leu Ala Glu Ile 145 150 Lys Ile Thr Giu Leu 140 Phe Arq Val Vai Leu 155 Asp Leu Ser Lys Arq Gin Ile Lys 160 Leu Ser Ser <210> <211> 1101 <212> DNA WO 00/31115 84 (213> Artificial Sequence <220> <223> Description of Artificial Sequence:Clone pAD922 PCT/AU99/01035 <400> actgatcaga gaccttqaaa tctcggattg gcaaaaatca tttagctcat gaagtccaaa gctaaaaaag aaaacattaa ttcaccgctc attqcaaata caagcaataa gaaaaaaacc aataatttca gaaaaaaatt ttggatattc gtaaattttc agtagtccgc aaagaagcaa acaaaaccag tcttaatttt tgctcgaaaa aaaaaataga acgaattttt catataatct aaatttaccg attatqaaga atgtaaaagc aaaacaattt acaaagcgct tcaataagga ttttttcctt aaaaaagtca atacaaatac aaggtttaaa aaacaaaatt aagccgaaaa tttatgaaga agcaaaaaaa ccatttqgcc aqgtaacttt agaaaattta taattcttat tggctatgaa acaaaaacag ccagctaaaa gcttaattta gttgacttcc agtaacaaca aattgccaaa tcttaatatt aattgaaaag aaaattattt agaaattgcc tctccaattt tctagaaaaa agaaaaagac aaaactttag gaatttcaag gcgcaaattt ttgctttatt aatgtgqcca caagcaaaag aaaataaagc aaaaaagagg tatattgacg gatctaaaaa tttcttgata tctgagattg tataaaagtt agtcaattaa caaaatcgta aaaaataaat ctcaatgaat ctttttatta accaaaaata atttcattaa ctaccgataa ttgaaaaatt aattatatqa tcgaatttga aagaaaaaqc cccaactcga aatttaatta attattcatt ggagaaatat aaaaattatt tgactttcga tccgtaccqa aaaaaactta.

attcatataa taaaagaaaa gaaaatatac tcttgaaatt tttctgqcaa.

aatcacagag acaaaaatta ttatttctat ctaccgcagt ttttttcatt gattgacaaa tgaatataaa ttttaaaaaa tttgttactt tgaaattatg taaaaaagat aattatcatt tcaagaaaaa taaaaaaaga actggcccaa ctcttggaaa 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1101 <210> 46 <211> 367 <212> PRT <213> Artificial Sequence WO 00/31115 <220> <223> Description of Artificial Sequence:Deduced protein sequence from clone pAD922 PCT/AU99/01035 <400> 46 Thr Asp Gin Ile Leu Ile Phe His Leu Ala Lys Thr Leu Asp Gin Lys Glu Lys Gly Asn Phe Glu Phe Tyr Leu Glu Ile Asp Leu Glu Met Leu 25 Gin Asp Phe Ile Asn Phe Asn Leu Ala Gin Ile Ser Glu Phe Phe Asn Ser Tyr 70 Trp Gin 40 Ser Arg Ile Glu Lys Ile Glu Glu Thr Asp Lys Ile Thr Glu Ala Lys Ile Asn 55 Leu Leu Tyr Phe Glu Lys Leu Gin Lys Leu 75 Phe Ser Ser Ser Tyr Asp Tyr Phe Tyr Glu 100 Asn Leu Gly Tyr Glu Asn Val Ala Lys Leu Tyr 90 Tyr Arg Gin Lys Gin Gin Ala 110 Val Gin Lys Lys Val Glu Phe Asp Tyr Arg Ser 115 120 Ala Lys Lys Asp Tyr 125 Glu Asp Gin Leu Lys Lys Ile Lys Gin Glu Lys Ala Phe Phe Ile Lys Thr Leu Asn WO 00131115 WO 0031115PCT/AU99/01035 140 Lys Ala Leu Asn Lys Lys Glu Phe Thr Ala Gin Asn Leu Leu Thr Ala Gin Leu Giu Ile Asp Lys 155 160 Ser Tyr Ile Asp Giu Phe Asn 170 175 Ala Leu Val Thr Thr Asp Leu 190 Tyr Glu Tyr Lys Asn Tyr 195 Ala Lys Phe 210 Ile Ala Asn Asn Ser Phe Phe Lys Gin Ala Ile Ile Asn Lys Giu Ile 205 Leu Asp Arg Arg 215 Asn Ile Leu Leu Leu Giu Lys Asn Leu 220 Leu Phe Glu le Met 240 Phe Ser Phe Leu Asn 225 Ser Giu Ile Glu Asn Asn Phe Lys Asp Lys Lys Asp 260 Leu Ile Arg Thr, 275 Ser Gin Ile Glu Lys Tyr Lys Ser Leu Thr Phe 250 255 Asn Thr Lys Leu Phe Ser Gin 270 Glu Lys Asn Tyr Gu Ile Ile Ile 280 Thr 265 Leu Asp Ile Gin Giy Leu Lys Giu 285 Tyr Gin Giu Lys Val Asn Phe Gin 300 Ile Ala Gin Asn Arg Lys Lys Thr 290 295 WO 00/31115 WO 0031115PCT/AU99/01 035 Thr Lys Phe Leu Gin Phe Lys Asn Lys Tyr 305 310 Ser Tyr Asn Lys Lys Arg 315 320 Ser Ser Pro Gin Ala Glu Asn Leu Giu 325 Lys Leu Ala Gin Lys Giu Ala Ilie Tyr 340 345 Ile Arg Lys Tyr Thr Ser Trp Lys Thr 355 360 Leu Asn Glu Leu Lys Glu 335 Giu Glu Giu Lys Asp Leu Phe 350 Lys Pro Glu Gin Lys Asn 365 <210> 47 <211> 438 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence:Clones pAD923 and pAD925 <400> 47 tataatttaa tcaagcattg tatgcattat gcttcaatgc atcgatgccc tatttaaatg aaaaagaaac aatttgatgc taaaaaaaaa cttgtaatcc ttggaggagt aatcaaaagq taatttgagg tattgttgtc gttaaaagtt cgcaattggt tcagggaaaa tcctgccgtt aaatttttaa ggtggcggcc gttctaataa gggccagcaa ttttcagatt ttagcaatta tgtcaaaaaa atgctgggat ctcttgataa aaggaattat tagcaatgat gagcccaaat atctaaaaat cgaagcagtt gaaaaaatta aactcgcgag ccaaattaaa tgataaggaa WO 00/31115 PCT/AU99/01035 88 aaatattcaa aattaatatt aaaggatttg aaaaaacagg aaaatttatt aattatcgag 420 gatttggtta gtgaactc 438 <210> 48 <211> 146 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence:Deduced protein sequence from clones pAD923 and pAD925 <400> 48 Tyr Asn Leu Lys Lys Glu Thr Asn Leu Arg Lys Phe Leu Met Ser Lys Asp Ala Ile Val Val Gly Gly Lys Ser Lys Asn Ser Ser Ile Glu Phe 25 Gly His Ala Gly Ile Glu Ala Val 40 Lys Val Val Leu lie Thr Leu Asp Lys 55 Ala Leu Leu Lys Lys Lys Leu Lys Lys Leu Ala Ser Met Pro Lys Gly Ile lie Thr Arg Glu 75 Lys Phe Ser Asp Leu Ala Met Cys Asn Pro Ala Ile Gly Gly Pro Ala 70 Ile Asp Ala Leu Gly Gly Val Gin Gly WO 00/31115 WO 0031115PCT/AU99/01 035 Ile Gin Ile Ile Arq Ala 115 Asp Leu Lys 130 Lys Tyr Leu Asn Giu Ser Lys Gly Pro Ala Val Leu Ala 100 105 110 Gin Ile Asp Lys Giu Lys Tyr Ser Lys Leu Ile Leu Lys 120 125 Lys Gin Giu Asn Leu Leu Ile Ile Giu Asp Leu Val Ser 135 140 Giu Leu 145 <210> <211> <212> <213> <220> <223> 49 1074

DNA

Artificial Sequence Description of Artificiai Sequence:Cione <400> 49 caaattgtac ctacaggaac aaccaatttg attgaacaac gaaattggcg tctaataaaa ttaaataatt cctaaaaatg aatcagaacc aaaaattaat aactcgccaa aaaatattga agaataattt attttattat attataaaat ctgaattttt agaaatttta tagtttttgc cgaatatcac gcttaacaac agtaattagt tgatattata taatatttat agacccaaaa aatcaaaaat qaacaaaaat aaagcgctta ctaaaaaata tttgcttttt cttaattata cccgatgagt attaatatca tttttttatg tagaaaaagc ttgcactgaa ttgattttct atcqcaatgg cagaagtttt tagtagttaa aaattggaaa taaaaaaata aaagaaaaat aaaaactaaa ttattatagt tgttttttta.

aattaatttt aaatttttgg aagcttaaaa WO 00/31115 WO 0031115PCT/AU99/01035 tataagcata qaccaagaaa ttaaaaattg taccccacaa tttttcaatt aaatatatta caacaaatta qgaatggtaa attaacctag tttgttaagg ttttaaacac ttaagaaaaa aaaatqttga ctggaattat atagqqgaac aaaatccaaa atttaattat aaaataaaag Ctcaagaagt aaaaaatgaa tttagcaaaa aattaaaaat aaaaattatg tttctatata aaaaaaaggt atttctaaaa agaaatttta gcataaaact tcttaatttc gaaaaaacaa aatcaccaag cagaaaatt t gcaattgact aacggaatat gatacaaatt cctgatttga aqaaaaaatc gaaaaaatta tttgctttaa ataaaaagtt attttatcag tagaactagt gctcaaattt atgagcgaaa atatqagaca ttttagtaga actttgaaat ttgacttaaa ttcaagaaaa tattttctaa ccataatttt taaaaccagt agagtcaaac ttacaataga gggttttgaa tggaggaatt tccgattttt tggtaaaaaa tgtcgattta ggaa 540 600 660 720 780 840 900 960 1020 1074 <210> <211> <212> <213> <220> <223> 358

PRT

Artificial Sequence Description of Artificial Sequenc:Deduced protein sequence from Clone <400> Gln Ile Val Gln Ser Glu Pro Giu Ile Leu Asn Gin Lys Phe Phe Leu 1 5 10 Cys Lys Lys Ile Leu Gin Glu Gin Lys Leu Ile Ser Phe Cys Giu Gin 25 Lys Leu Giu Lys Ala Lys Lys Asn Asn Gin Phe Giu Leu Ala Asn Giu 40 Tyr His Lys Ala Leu Ile Ala Leu Lys Lys Thr Lys Ile Giu Gin Gin WO 00/31115 WO 0031115PCT/AU99/01035 Asn Ile Glu Leu Asn Asn Leu Lys Asn Ile Asp Phe Leu Tyr Tyr Ser 70 75 Giu Ile Gly Giu Asn Asn Leu Val Ile Gly Vai Phe Leu Ser Asn Lys Asn Phe 100 105 Tyr Thr Giu Vai Leu Ile Asn Phe Leu 115 120 Ser Phe Ala Phe Tyr Arq Asn 90 Ile Ile Asp Ile Ile Leu Asn 110 Lys Ile Asn Asn Asn Tyr Ile Tyr 130 Pro Asp Giu Leu Vai Vai Lys Asn Phe 135 T rp 140 Pro Lys Asn Ala Giu 145 Phe Leu Asp Pro Lys Ile Asn Ile Lys 150 Gly Lys Ser Leu Lys 160 Tyr Lys His Ile Leu Asn Thr Leu Ala Lys 165 170 Asp Gin Giu Ile Lys Lys 185 Asn His Gin Asp Phe Ile 175 Ser His Asn Lys Ile Lys Asn Gin Lys 190 Val Giu Lys Ile Leu Giu 195 Leu Val Lys Thr Ser Leu Lys Ile Glu Asn 200 205 Ile Met Aia Ile Asp Cys Ser Asn Leu Giu Ser Asn Tyr Pro Thr Thr WO 00/31115 PCT/AU99/01035 210 Gly Ile Ile Phe Tyr Ile Asn Gly Ile Tyr Glu 225 230 235 Phe Phe Asn Tyr Arg Gly Thr Lys Lys Gly Asp 245 250 Arg Asn Tyr Asn Arg 240 Thr Asn Tyr Met Arg 255 Gin Gly Phe Glu Lys Tyr Ile Lys Asn Pro 260 265 Lys Phe Leu Lys Pro Asp 270 Leu Ile Leu Val Asp Gly Gly Ile Gin Gin Ile Asn Leu Ile Ile Glu 275 280 285 Ile Leu Arg Lys Asn His Phe Glu Ile Pro Ile Phe Gly Met Val Lys 290 295 300 Asn Lys Arg His Lys Thr Glu Lys Ile Ile Asp Leu Asn Gly Lys Lys 305 310 315 320 Ile Asn Leu Ala Gin Glu Val Leu Asn Phe Phe Ala Leu Ile Gin Glu 325 330 335 Asn Val Asp Leu Phe Val Lys Glu Lys Met Lys Lys Lys Gin Ile Lys 340 345 350 Ser Leu Phe Ser Lys Glu 355 <210> 51 WO 00131115 PCT/AU99/01035 <211> <212> <213> <220> <223> 93 282

DNA

Artificial Sequence Description of Artificial Sequence:Clone pAD951 <400> 51 tcgaatataa atttttaaag aacaataata gatcatgata gttagtcgtt ctgacaaaac aggaaaactt ttaaaaattt ctaacaataa aaatacttta ttgttggagt ttttgatcct gaaaaagatg acgaaaatat tgctattttt 120 ttgaaaaata ttctagtgaa ttacttccaa tagctgctgt ggtttatttt 180 atttatataa taatattaat gaatttttaa ataaatatag caaaccgggc 240 attactcgac aaatggcggc cggatcaaat tc 282 <210> <211> <212> <213> <220> <223> 52 94

PRT

Artificial Sequence Description of Artificial Sequence:Deduced protein sequence from clone pAD951 <400> 52 Ser Asn Ile Thr Asp Lys Thr Gly Lys Leu Leu Lys Ile Ser Asn Asn 1 5 10 Lys Asn Thr Leu Ile Phe Lys Val Val Gly Val Phe Asp Pro Glu Lys 25 WO 00/31115 PCT/AU99/01035 94 Asp Asp Giu Asn Ile Ala Ile Phe Asn Asn Asn Ile Giu Lys Tyr Ser 40 Ser Glu Leu Leu Pro Ile Leu Tyr Asn Asn Ile Asn 70 Val Ser Arg Tyr Tyr Ser Ala Ala Val Val Tyr The Asp His Asp Asn 55 Giu Phe Leu Asn Lys Tyr Ser Lys Pro Giy 75 Thr Asn Gly Gly Ar Ile Lys Phe <210> <211> <212> <213> <220> <223> 53 438

DNA

Artificial Sequence Description of Artificial Sequence:Clone pAD977 <400> 53 attttaatta attqcactaa ttatgagata gataattttt caaaaagttg gctcttgata ccgaataaaa gaccaaataa ataattcaat caacaaatga aaaatcccga caaaaccaga cattggccag acattttcaa caatttttat ttgaaatt tgaatataaq aaacataatt tttaqatttq aactgaaatt attagagtac ggaagatqct cgcaagtcac gaattagacc ttcgaagaca ctaaattctt agctcaaaaa aaaaatttag tttgaaattt cacattcctq caaaccaqtt ctttggcaaa taataaaaaa atttatctga atatttgatq acagtqattt aaaaaataaa aaqaaaacat caacataact gtataaaatt gggcgaaaaa tttagatgaa actttcaaaa accgatgttt WO 00/31115 PCT/AU99/01035 <210> <211> <212> <213> <220> <223> 54 146

PRT

Artificial Sequence Description of Artificial Sequence:Deduced protein sequence from clone pAD977 <400> 54 Ile Leu Ile Asn Asn Ser Ile Glu Tyr Lys Glu Leu Asp Pro Asn Gin 1 5 10 Leu Arg Lys His Ile Ala Leu Thr Thr Asn Glu Asn Ile 25 Asp Thr Leu Ala Asn Asn Ile Thr Leu Trp Asp Lys Asn 40 Asp Leu Leu Asn Ser Leu Ile Lys Lys Tyr Lys Ile Asp 55 Lys Pro Glu Thr Glu lie Ser Ser Lys Asn Leu Ser Glu 70 75 Ile Phe Glu Pro Asp Leu Asn Phe Ser Gly Glu Lys Gin Lys Val Ala Leu Ala Arg Leu Glu Tyr Lys Asn Leu Asp Ile Trp 90 Cys Leu Asp Glu Ala Leu Asp Asn Ile Phe Lys Glu Asp Ala Phe Glu 100 105 110 WO 00/31115 PCT/AU99/O 1035 96 Ile Tyr Ser Asp Leu Leu Ser Lys Pro Asn Lys Thr Ile Phe Ile Ala 115 120 125 Ser His 130 Glu Ile 145 His Ile Pro Glu Lys Ile Lys Pro Met Phe Asp Gin Ile Ile 135 140 <210> <211> <212> <213> <220> <223> 400

DNA

Artificial Sequence Description of Artificial Sequence:Clone pAD983 <400> tcaacgggct ttatttaatt tctgaaataa ggccatctaa ataaaaacat tttgcagttt tttgttcaaa <210> 56 <211> 133 <212> PRT gccaaattga atagctttta caattttctt taattggqac caggatgaat ttgcaggaat cagctcaaat acctgacaaa ttattcagga tcttcaaaca aqttttttgt tgcgcttgca tctatttcca aqttttaatt ccgctggtaa attctaagca gcagqggctg atttttttca tcaacaattt aaaacttata ttgcaaaatt aaaaatgqgt tagttttagg atattaatgt ctagccttaa taaaatttat atgccgqcqg tatqggtgtt atttttttct tccagtttqa t tat at t tca tcctttagta ttctaatgcc 120 180 240 300 360 400 WO 00/31115 97 <213> Artificial Sequence <220> <223> Description of Artificial Sequence:Deduced protein sequence from clone pAD983 PCT/AU99/01035 <400> 56 Ser Thr Gly Cys Gin Ile Glu Pro Asp Lys Pro Leu Val Lys 1 5 10 Lys Trp Val Met Gly Val Leu Phe Asn Tyr Ser Phe Tyr Tyr Ser Gly Ile Leu 25 Ser Ile Val Leu Gly Phe Phe Ser Ser Glu Ile Thr Ile 40 Gin Thr Ala Gly Ala Asp Ile Asn Val Pro Val Trp Gly 55 Ile Gly Thr Val Phe Cys Ile Phe Phe Thr Ser Leu Asn 70 75 Phe Phe Leu His Leu lie Tyr lie Ser lie Lys Thr Ser Gly Trp Ile Ala Leu Ala Ser Thr lie Leu Lys Phe 90 lie Pro Leu Val Phe Ala Val Phe Ala Gly Ile Leu Phe Pro Lys Thr 100 105 110 Tyr Asn Ala Gly Gly Ser Asn Ala Phe Val Gin Thr Ala Gin Ile Val 115 120 125 WO 00/31115 WO 0031115PCT/AU99/01 035 Leu Ile Leu Gin Asn 130 <210> 57 <211> 522 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence:Cione pAD984 <400> 57 ttaaaaagtg agtaaaaaag aaagataaaa gaaatttctc tcaaatttaa ttaqagaata agtcaaaaaa agaaaaaata aaccaggaga aaaaccaaaa atatcgaaat tgcgaagtga aactaaaaga aactagaaca aatttaacga cacgagaaat aaacaggaaa aaaaattcaa agaaacagca aaccaattta cattcttqca agcaaatgcc taaggacgaa cttggaaqaa aaaggaaaaa aatcagattt gattagaaga aatttaaata agaaatgaaa aatgatgatg aacctacaaa attaatgaaa gaaaaaaata attgaaaaag aagaggaaaa agaattaaat ctacttttac ttggcaaatt agataaaggc atgtcaagtt aagaccgtaa attcaattca agctagaaat ttttaaggag ta tcaaacaatt tcttgatgaa gatgaggagt agaaqaaatt aattagttat aaatgctgta taaatgggga caaattaata <210> 58 <211> 174 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Deduced protein WO 00/31115 PCT/AU99/01035 99 sequence from clone pAD984 <400> 58 Leu Lys Ser Glu Asn Gin Lys Glu Thr Ala Asn Leu Asn Thr Thr Phe 1 5 10 Thr Gin Thr Ile Ser Lys Lys Asp Ile Giu Ile Thr Asn Leu Arg Asn 25 Giu Ile Gly Lys Phe Leu Asp Glu Lys Asp Lys Met Arg Ser Asp Ile 40 Leu. Ala Asn Asp Asp Glu Ile Lys Ala Met Arg Ser Giu Ile Ser Gin 55 Leu Lys Giu Ala Asn Ala Asn Leu Gin Asn Val Lys 70 75 Leu. Glu Glu Ile Ser Asn Leu Lys Leu Giu. His Lys Asp Glu Ile Asn Giu Lys Asp Arg 90 Lys Ile Ser Tyr Leu Glu Asn Lys Phe Asn Asp Leu Giu.

100 105 Asn Asn Ser Ile Gin Asn Aia Vai Ser Gin Lys Thr Arg 115 120 125 Glu. Lys Ilie Giu Lys Giu Leu. Gu Ile Lys Trp Giy Arg 130 135 140 Giu Giu Lys 110 Giu Ile Lys Lys Asn Lys WO 00/31115 PCT/AU99/01035 100 Thr Gly Lys Ile Arq Phe Lys Arg Lys Ile Leu Arq Ser Lys Leu Ile 145 150 155 160 Asn Gin Giu Lys Lys Phe Lys Ile Arq Arg Arg Ile Lys Leu 165 170 <210> <211> <212> <213> <220> <223> 59 633

DNA

Artificiai Sequence Description of Artificial Sequence:Cione pAD994 <400> 59 attaaacact aaattagatt tttaatttaa tttacaatqa ttggatattg attgagctaa gaaaaattag aaatttgata aaaaaaattc gaagttcaaa aaaaaaattt tttttaaaag ttgaaaattt ccgctttttc atttcattgt gggctggatc caatttctga atttgaaatt taataactgc attttccaaa attgtaaaaa ttatcgcatt atttgaaatg agaaaaatac tggtgatatt tggtttagta aggttttcct gtcaatgcaa caatttaatt caqagcagtq aacgctttta ttgtaattat taaacaagtt tataaacgat gtaaatttaa ctttgaaaag aataataaag tcaattcctt aaaagat qcc tgcaaaccaq gcaaatttgg gcttttatta aaaatcatta t ct tagttcagga t tgaatt tag aqqgaatttt aaaataaaaa ttttgattac agtttttaaa ttcaagaaat aaaagcttga aagggcccaa aagttaataa attctttcct taataaaaac tgagtcaatt attaaaaatt aaacccagaa ggatqtttct taatccacaa gaaaattaca agtttttaat taatataaat <210> <211> <212> WO 00/31115 WO 0031115PCT/AU99/01 035 <213> Artificial Sequence <220> <223> Description of Artificial Sequence:Deduced protein sequence from clone pAD994 <400> Ile Lys His Phe Phe Lys Arg Phe Glu Met Tyr Lys Arg Leu Val Gln 1 5 10 Glu Phe Phe Pro Lys Leu Asp Phe Glu Asn Leu Glu Lys 25 Tyr Val Asn Leu Ile Glu Phe Ser Asn Lys Asn Phe Asn Leu Thr Ala Phe Ser Gly Asp Ile Leu Trp Lys Glu Gly Ile Phe Glu Ser Ile Phe 55 Thr Met Asn Phe Ile Val Gly Leu Val Asn Asn Lys Glu Asn Lys Lys Leu Lys Ile 70 75 Leu Asp Ile Gly Ala Gly Ser Gly Phe Pro Ser Ile Pro Phe Leu Ile 90 Thr Asn Pro Glu Ile Glu Leu Thr Ile Ser Glu Ser Met Gln Lys Arg 100 105 110 Cys Gln Phe Leu Lys Asp Val Ser Glu Lys Leu Asp Leu Lys Phe Asn 115 120 125 WO 00/31115 PCT/AU99/01 035 102 Leu Ile Cys Lys Pro Val Gin Giu Ile Asn Pro 130 135 Gin Lys Phe Asp Ile 140 Thr Ala Arg Ala Val Ala Asn Leu Giu 150 Leu Giu Lys Ile Lys Lys Ile His Lys Val Phe Asn 180 Phe Pro Lys Thr Leu 165 Giu Vai Gin Asn Cys 185 Leu 170 Ala Phe Ile Lys Gly Pro 175 Lys Asn Cys Asn Tyr Lys Ile 190 Lys Ile Phe Ile Ala Phe Lys 205 Ile Lys Val Asn Asn Asn Ile Asn Lys 195 200 Gin Vai Ser 210 <210> <211> <212> <213> <220> <223> 61 537

DNA

Artificial Sequence Description of Artificial Sequence:Cione pADlOOS <400> 61 atqaaaaaat tattagttat tttgcttgat aaattccagg atattgaact tacaactttt atttccctga ttaaaaaagc agaaattttt acagatattg aattttttaa ccctaaaaat 120 WO 00/31115 WO 0031115PCT/AU99/01 035 103 aataaattaq tcagatgact gattcaaaat atttgtgatg tcatatccaa gaccgcaatt attgaaaagc taataggtca ttgatgctqt caattcaact caccaaatgc accaacataa atatttcggc tgggatcaaa attcggagtt ttttattccg tgtgagcqag aatttttgaa ttccaaaatt aagaaatqca agagttagct gtatcaattc gqgggttttg ttttttgcgc ctaaaattag agactaagac gcaagttcgg caaaaaatta aagcacataa cggcccaatt aaaacaaaca c aga a aatt a aagattcgtt cagattttgc gaaatggatt tcactggaaa attccgcaag tatttttgcc tcaatttagt agtaactatt tttcgttgta ttatctt 180 240 300 360 420 480 537 <210> <211> <212> <213> <220> <223> 62 179

PRT

Artificial Sequence Description of Artificial Sequence:Deduced protein sequence from clone pAD1OOS <400> 62 Met Lys Lys Leu Leu Val Ile Leu Leu Asp Lys Phe Gin Asp 1 5 10 Ile Glu Leu Thr Thr Ile Giu Phe Gly Val Val Phe Ile Ser Leu Ile Phe Asn Pro Lys Asn 40 Ser Ile Gin Ala His 55 Lys Lys Ala Glu Ile Phe Thr Asp 25 Asn Lys Leu Val Ile Gly Gin Phe Asn His Trp Lys Ser Asp Asp Phe WO 00131115 WO 0031115PCT/AU99/01 035 104 Asp Ala Val Phe Ile Pro Gly Giy Phe Ala Ala Gin Leu Phe Arg Lys 70 75 Gin Leu Val Ser Giu The Phe Ala Gin Asn Lys Asp Ser Lys Ser His Ile Phe Leu Ala Giu 115 Lys Ile Arg 130 Ile Cys Asp Ala Pro Asn Ala Ile Phe Giu Leu Lys 105 110 Ser Tyr Pro Asn Gin His Asn Ser 125 Asn Tyr Gin Phe Leu Arg Gin Asp 135 Ser Leu Vai Thr Ile Asp Arg Asn Tyr 140 Ile Ser Ala Arq Asn Ala 145 150 Ile Glu Lys Leu Gly Ser 165 Ala Ser Ser Aia Asp Phe Ala Phe Val Val 155 160 Lys Glu Leu Ala Gin Lys Ile Arg Asn Giy 170 175 Phe Tyr Leu <210> <211> <212> <213> <220> 63 486

DNA

Artificial Sequence WO 00/31115 105 <223> Description of Artificial Sequence:Cione pADlO16 PCT/AU99/01 035 <400> 63 ttaatcgttt ct t aacggt a gccccaqgaa ggatcagttc tctttatttt ataattctta gttgtctcaa tttttatcca agctca.

ttgcttatat aagcgggaat ccgttgtaaa ttgttggttg ttatgctttt gaacaatttt attatttaaa attggaacaa gatgttggtt aagtggttat tcaagcagtt attaattgtg tgqaattatt tgcaattggc tcaaaggcaa taattacgct gtaatgaatt ttaggtcact aactggqta aaaatttcqc qccccttatt qgaacaatgc aaagctgtta tattcctttt gaggttttgc tttttccaaa tcacaattgg ataaatatac caccaactta agattatttt tttcacaagt gcaggaatta ctctgccgga tgctaatqaa tcgtggaatt agtaattttg tgccgggttt aattcaacca tttccccttt tttggtcaag 120 180 240 300 360 420 480 486 <210> <211> <212> <213> <220> <223> 64 162

PRT

Artificial Sequence Description of Artificial Sequence:Deduced protein sequence from clone pAD1Ol6 <400> 64 Leu Ile Val Phe Ala Tyr Met Met Leu Val Val Met Asn Trp Gly Phe 1 5 10 Ala Ser Ala Gly Leu Asn Gly Lys Ala Gly Ile Ser Gly Tyr Leu Gly 25 His Phe Phe Pro Asn Ala Asn Glu Ala Pro Gly Thr Val Val Asn Gln WO 00/31115 WO 0031115PCT/AU99/01 035 106 Ala Val Asn Trp Gly Ile Thr Ile Gly Arg Gly Ile Gly Ser Vai Leu 55 Val Gly Trp Leu Ile Val Lys Ile Ser His Lys Tyr Thr Val Ile Leu 70 75 Ser Leu Phe Phe Met Leu Phe Gly Ile Ile Ala Pro Tyr Ser Pro Thr 90 Ile Ile Leu Arg Thr Ile Phe Ala Ile Gly Gly Thr 105 110 Tyr Ala Gly Met Gin Ile 115 Ile Leu Ile Gin Arg Gin 130 Trp Asn 145 Lys Ala Vai Ile Ser 135 Asn Asn Tyr Ala Tyr 150 Pro Val Val Ser Asn Tyr Leu Asn Gin 120 125 Gin Vai Phe Pro Phe Phe Leu Ser Asn 140 Ser Phe Cys Arg Asn Tyr Leu Val Lys 155 160 Ser Ser <210> <211> 456 <212> DNA <213> Artificial Sequence WO 00/31115 107 <220> <223> Description of Artificial Sequence:Clone pAD1O20 PCT/AU99/0 1035 <400> tctgatcgtt cctatttttg aagatgaata qaagatgcac gttgtagaaa tctggtgctt tatgacttta tcttattatt ttccaqttac aaaaattcaa gttgagaccg aaaaaatagt aagcatttag attcaattgg ctataaacgc ctgataaaaa aagtttaqaa aaaagcaaaa tttagttcca gcttggggca ccaaagatga gggttcatat agttcatact ccagaattat aaattatttc gaaaaatttt ttagttgaga atcaaaccat attgactatc gggcttgaaa ttagcacatg cataat aatttgtcgc atcaagcaaa caaaaaataa taggtcaaga attatgttga aaaaatatat aattaggtca taattcagcg atttgggact ttattctgtt atataaagat ttcaaaaaga tttaatgaat ttcgctccat 120 180 240 300 360 420 456 <210> <211> <212> <213> <220> <223> 66 152

PRT

Artificial Sequence Description of Artificial Sequence: Deduced protein sequence from clone pAD1O2O <400> 66 Ser Asp Arg Phe Pro Val Thr Ser Leu Glu Lys Leu Phe Gin Phe Val 1 5 10 Ala Asn Ser Ala Pro Ile Phe Glu Lys Phe Lys Lys Ala Lys Glu Lys 25 WO 00131115 PCT/AU99/01035 108 Phe Tyr Gin Ala Lys Phe Gly Thr Lys Met Asn Ser Trp Asp Arg Leu 40 Val Pro Leu Val Giu Thr Lys Asn Asn Tyr Ser Val Glu Asp Ala Gin 55 Lys Ile Val Leu Gly Ala Ile Lys Pro Leu Gly Gin Glu Tyr Lys Asp 70 75 Phe Ser Gin Arg Trp Ile Asp Tyr His Tyr Val Val Val Glu Lys Asp Ser Lys Giu Lys Lys 115 Ser Gly Ala Tyr Ile Leu Met Tyr Ser Ile Gly Gly Ser Tyr Gly Leu 105 110 Asn Tyr Asp Phe Thr Ile Asn Aia Val 120 125 Gly His Ser Leu His Ser Tyr Tyr Ser 140 His T1 E2 Asp Ll 145 <210> <211> <212> <213> <220> 30 Leu Ala His Giu Leu 135 is Asn Gin Asn Tyr His Asn 150 67 576

DNA

Artificial Sequence WO 00/31115 109 <223> Description of Artificial Sequence:Clone pAD1027 PCT/A U99/0 1035 <400> 67 gaattaatta ccttggatat ctaattaata aatagtgaaa.

gaaataaatg aagtttgatt ctacccgagc actagtaaaa ccagaaaatc ggtgatgaaa gggaaaattt caacaacaaa ctaataaaat tacctgcttt ttgatatctc gggacctaaa.

caaaatctga aactagtaaa ctgatttaag atgctataga atcacttgca aaattttgaa aaaaaattat agaacttaat aaagtgagtc accagacctg ggtattctta tataataaat gttaqttagc aaattttaaa aaatcatttt aacttatttg atcacaaaca aatttgctaa caacaaaaac aatcagtatg ctaagaaaag aaaattaagg caactttatt gggatt atqttgataa attatgtaca taaattttaa aagatgataa taattaaaat cccggatttt atgaaaattc gatttaacaa tacttgattt aaataataat aagcgaqcat aatcaaaaaa aattcggctt tttaagtttt tgcacaaaat agcagcgtga tqqattagac tggcaaaatt <210> <211> <212> <213> <220> <223> 68 192

PRT

Artificial Sequence Description of Artificial Sequence: Deduced protein sequence from clone pAD1027 <400> 68 Glu Leu Ile Arg Glu Asn Leu Ser Leu Ala Lys Ser Phe Tyr Val Asp 1 .5 10 Lys Asn Asn Asn Pro Trp Ile Ser Thr Thr Lys Asn Phe Glu Asn Leu 25 WO 00/31115 PCT/AU99/01035 110 Phe Asp Tyr Val Gin Ser Giu His Leu Ile Asn Thr Asn Lys Ile Lys Asn Tyr Ile Thr Asn Ile Asn Phe Lys Ile Lys Lys Asn Ser Giu Ile 55 Ala Leu Giu Giu Ile Asn Val Leu Asn Asn Leu Leu Lys Asp Asp Lys Ile Arg Leu 75 Asp Ile Ser Lys Trp Val Gin Gin Lys Leu Ile Lys 90 Lys Phe Asp Trp Asp Leu Lys Pro Asp Leu Asn Gin 105 110 Phe Ala Gin Asn Leu Pro Giu Pro Lys Ser Giu Vai 120 125 Ile Leu Ser Phe 100 Tyr Aia Arg Ile 115 Phe Leu Leu Arg Lys Asp 130 Leu Vai Asn Ile Ile Asn 145 150 Asn Ser Ala Ala Trp Thr Ser Lys Lys 140 Asn Asn Gly Leu Asp 160 Lys Ile Lys Gly Phe 155 Pro Glu Asn Pro Asp Leu Arg Leu Val 165 Phe Giy Lys Ile Giy Asp Giu Asn Aia 180 185 Ser Gin Leu Tyr Leu Leu Asp 170 175 Ile Glu Asn Phe Lys Gly Ile 190 WO 00/31115 WO 0031115PCT/AU99/O1 035 <210> 69 <211> 351 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence:Clone pAD1037 <400> 69 catatgttaa aaaaaatcag ggcgcctttt tctggcgggg ttaagaaata attttagcct ttgaagtttt taatttataa tgaaccgttt caattttaag acccttcttt taattatttt aataattcac cttaataaca ttttattaaa ttttgttgtc tcaqtgacat aatttacatg taccgtcaag accctgattt acaggctgac ataattgagt ttttctcttt aacattaagc ttcagtatgg tagtgccgat taataccatt tagttccaga ttttgtttgc qccgt agatc ccaaagtatt tattacagtt 120 ttttaatgtt 180 atttatccat 240 tttaggaatt 300 c 351 <210> <211> <212> <213> <220> <223> 117

PRT

Artificial Sequence Description of Artificial Sequence: Deduced protein sequence from clone pAD1O37 <400> His Met Leu Ile Glu Val Leu Ile Ile His Tyr Arg Gln Val Gln Tyr WO 00/31115 WO 0031115PCT/AU99/01 035 Gly Gin Ser Ile Lys Lys Ser Val Ile Tyr Asn Leu Ile Thr Thr Leu 25 Ile Leu Val Pro Ile Ile Thr Vai Gly Ala Phe Leu Asn Arg Phe Phe 40 Ile Lys Thr Gly Trp Leu Ile Pro Phe Phe Asn Val Ser Gly Gly Ala 55 Ile Leu Ser Phe Val Val Ile Ile Glu Leu Val Pro Giu. Phe Ile His 70 75 Leu. Arg Asn Asn Pro Ser Phe Gin Trp His Phe Ser Leu. Phe Leu Phe 90 Ala Leu Giy Ile Ile Leu Ala Leu Ile Ile Leu Ile Tyr Met Asn Ile 100 105 110 Lys Arq Arq Arg Ser 115 <210> 71 <211> 1776 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence:Clone pAD1O38 WO 00/31115 WO 0031115PCT/AU99/01 035 <400> 71 gaattcgaaa attattttta atggattttg gctaccacaa atgcaaaaag attaaggaaa gctacaaaag ttagtcgatg gaaaaagcaa gataattcag qatcaatggq gaaaaactaa gagattaaat aggcaggaaa acaaaaattc aaaaatttaq attttgcgqt atgggaccaa aataataaaa tcaaagctaa aataaagtaa ccggaagtaa tcacaaaaag aaaattcttg ttaaagccag gatataattt aataattttg aacgaattaa ttqatgaaat ccaatatcct attccgaata tagaaatttt ggctagaaaa cggcaaatcg aagctgctgc agcgcgagct aattaaaatc atcaatcaaa aatatcaaca acggaaaaat tttccaatgt cgattgaaaa caaaatcgct ttaaagctaa ctggggt ggg accaaataat ttggggctcc gactaaatcc tcaacatttt taaattttcg agggtaaaaa aatttctcaa ttgaaattat agattgaatt ggcaattttg tcaccttcta aaaaccaatt tcgcttatat agagccttca tttccatcaa ttacattttt ttctttaaaa aatttattta caaaattgaa aaaatcagcc aaattactta tcccaaaata tctagacgaa acttttagaa taagggtcaa aattaatgat aaaaactqaa ccgtcttgat tccgqqttat ttattcgatt tttacaaatt caatacaatt qatgaatgaa ccgaattgat tgaacttgaa tgaaaaatca caagaaattg attggaacag atggctcgcg atcgaaaaag gttattgata qtaaaaatta gaccgctttc gttgaaatca aaaatggaag aatcttgaaa tctgaaatcq atggaacaag agtaaaaaat agtcagatcg tcagaaattc aatcaggcaa gaatcccgcc cttgctagag atgtcagaat attggttttg attttgcttg cttgataatg ataattatga attgaggcaa gaaattatcg ctaaagqatt gtcaaaaatt agcaaaattc gatcttctqg gacagattaa atggcgccct caattaatat aggattctgc tacctgataa actaccaacc aaattaactc atgaagtaaa ctagcttatc gagactacca taggcgaaat caaaggttgt aaaaatattt ttaaggctgt caatttcatc ctttagctct atatggaaaa aacaaggtqg atgaaattga gtgaaat tgt cctcaaatat aaaaggaact tatttaatcc tgcaaaatcg gaattttaga cgatcaagtt gactgattca attaatcggg tgaaaqaaga tttacgggga tcttgttttt agctatcgat agaaaaactt gcaaaaacaa aaaattacaa ccaggaactt aaaagccgcc taaagcaaqa ctctaattga gaatttagaa ttcagatgcg atttttcttt taatttattt acatagtgtt taatctaaca aaaagctcat tgatagtaag cggtqctaat tttaagatat tttaaattat tctaaaggaa gtttggatat 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 gataaaaatt ttggtgccag gccaattaag cgctttatta agaaagaaat tgaaaatttt 1680 WO 00/31115 PCT/AU99/01 035 114 qttgccaaaa aaatagtqgc cgaagaaatt ttaaaagata aaaaatacaa tttatctttt 1740 aaaaatqata aattgcatct taatgaaagc qaaaat 1776 <210> <211> <212> <213> <220> <223> 72 592

PRT

Artificial Sequence Description of Artificial Sequence:Deduced protein sequence from clone pAD1O38 <400> 72 Glu Phe 1 Ser Asp Glu Lys Arg Ile Lys Ala Ile Leu Gin Glu Ile Glu Gin Asn 5 10 Gin Val Ile Ile Phe Ile Asp Giu Ile His Leu Leu Ile Gly 25 Thr Gly Ser Ser Gly Thr Asp Ser Met Asp Phe Ala Asn Ile Leu Lys 40 Pro Ile Met Ala Arg Gly Gin Ile Lys Leu Ile Gly Ala Thr Thr Asn 55 Ser Giu Tyr Arg Leu Tyr Ile Giu Lys Asp Giy Ala Leu Glu Arq Arg 70 75 Met Gin Lys Val Glu Ile Leu Giu Pro Ser Vai Ile Asp Thr Ile Asn 90 WO 00/31115 WO 0031115PCT/AU99/01035 115 Ile Leu Arg Ile Lys Asp 115 Giy Ile Lys Giu Arg Leu Giu Asn Phe His 100 105 Gin Vai Lys 110 Asn Arg Tyr Ser Aia Leu Vai Phe Aia Thr Lys Ala 120 Ala 125 Ile Phe 130 Asp Arg Phe Leu Pro Asp Lys Ala Ile Asp 135 140 Val Glu Ile Asn Tyr Gin 155 Leu Val Asp Glu Pro Glu Lys Leu Ala 145 Ala Ala Ser Leu Giu Lys Ala Lys Glu Leu Ile Tyr Leu Lys Met Glu Giu Ile Asn 170 175 Ser Gin Lys Gin 180 Asp Asn Ser Giu Leu Lys Ser Lys Ile 185 Giu Asn Leu 190 Ser Lys Lys Giu Asn Glu 195 Val Lys Lys Leu Gin Asp Gin Trp Asp Gin 200 205 Leu Ser Gin Giu Leu Giu 220 Ser Ala 210 Tyr Gin 225 Ser Giu Ile Ala Lys Leu Lys Gin Asn Tyr Leu 230 Met Giu Gin Gly Asp Tyr Gin Lys Ala Ala 235 240 Ile Pro Lys Ile Ser Lys Lys Leu Gly Glu 250 255 Glu Ile Lys Tyr Gly Lys 245 WO 00131115 WO 0031115PCT/AU99/O1 035 116 Ile Lys Ala Arg Arg Gin Giu Ile Ser Asn Val Leu Asp 260 265 Glu Ser Gin 270 Glu Lys Leu Ile Ala Lys Val Val Ser Asn Trp Thr Lys Ile Pro 275 280 Leu Giu Ser Giu Ile Gin Lys Tyr Leu Asn Leu Giu 290 295 300 Lys Ser Leu Lys Gly Gin Asn Gin Ala Ile Lys Ala 305 310 315 Lys Asn Leu Ala Val Ser Asp Ile Leu Arg Phe Lys Ala Lys Ile Asn Asp Giu Ser Arg Pro Ile Ser 325 330 335 Ser Phe Phe Phe Met Gly Pro Thr Gly Val Gly Lys Thr 340 345 Giu Leu Ala 350 Ile Ile Arq Arg Ala Leu Ala Leu Asn Leu Phe Asn Asn Lys Asn 355 360 Leu Asp Met Ser Glu Tyr Met Giu Lys His Ser Val 370 375 380 Gin 365 Ser Lys Leu Ile Gi y 385 Ala Pro Pro Gly Tyr Ile Gly Phe Giu Gin Giy 390 395 Gly Asn Leu Asn Lys Val Arg Leu Asn Pro Tyr Ser Ile Ile Leu Leu Asp Giu Ile 405 410 415 Giu Lys Ala His Pro Giu Vai Ile Asn Ile Phe Leu Gin Ile Leu Asp WO 00/31115 WO 0031115PCT/AU99/01 035 117 Asn Gly Giu 435 Ile Val Asp Ser Lys Ser Gin Lys Vai Asn 440 445 Phe Arg Asn Thr Ile 450 Giy Lys 465 Leu Lys Ile Ile Met Thr Ser Asn Ile Gly Ala Asn Lys 455 460 Ile Leu Giu Lys Met Asn Pro Glu Phe 485 Giu Ile Giu Ala Lys Lys Glu Leu Leu Arg Tyr 470 475 480 Leu Asn Arg Ile Asp Giu Ile Ile Vai Phe Asn 490 495 Pro Leu Asn Asp Ile Ile Phe Glu Ile Ile Giu Leu Giu Leu Lys 505 510 Asp Leu Gin 515 Lys Ser Vai 530 Asn Arg Leu Lys Giu Asn Asn Phe Glu Ile 520 525 Leu Glu Phe Gly Tyr Asp 540 Giu Phe Giu Lys Asn Phe Lys Asn Trp Giy Aia Arq Pro Ile Lys Arg Phe Ile Lys Lys Glu Ile Giu Asn Phe 545 550 555 560 Vai Ala Lys Lys Ile Val Aia Giu Giu Ile Leu Lys Asp Lys Lys Tyr 565 570 575 Asn Leu Ser Phe Lys Asn Asp Lys Leu His Leu Asn Glu Ser Giu Asn WO 00/31115 WO 0031115PCT/AU99/O1 035 118 <210> <211> <212> <213> <220> <223> 73 796

DNA

Artificial Sequence Description of Artificial Sequence:Clone pAD104O <400> 73 atggtaaaat acgatttttt aatgaaaatt aatatctttt atgaaacttg gaaaattcaa caattttatg ggtttatttt tttqtaatta caagcaattt ctacaaaaca ttaaaattta taactttttt tagattttat ccgaaaaaat gtgggtttta caaatttatt tttatataaa caacttcttt cggattttaa tttcaaattt tctcattgtt ttgggtagcc gaatcaagca aacaaatttt ctattcggaa gtcctttttg ctqgatttta actatctaaa caattcttta taaattcgaa tggtatattt aataatcaca atcctttgaa gctccttatt acaacatctt tattttaaag ttqqaaaaaa attgaaaata caaactcttt gcgttaatta aaattaacca agcacttatc tttataatta tccgacttaa attggaattg attgattata ttatttttac ttttaggggt ggtttttgat cgacttacaa caatagaaaa ccataatttt ttqtcggtgt aacttcaatc ttttaacttt ataaaagaaa tttcattttt cattttggac tttgattttt ttttattcta tagaattttt ctttcataag aaaatataac aagcgtttca gatgacttta tataaccttt cgcaaaattg gccgcaatta caagaaaaat taatgaatat actttattat aaaaccttaa attctgatga tgaataacta tccaataatt aaaaataata gagaaaaata ttttctccgc atagct <210> 74 WO 00131115 WO 0031115PCT/AU99/O1 035 119 <211> <212> <213> <220> <223> 265

PRT

Artificial Sequence Description of Artificial Sequence:Deciuced protein sequence from clone pAD1O40 <400> 74 Met Val Lys Ser Thr Lys His Phe Lys Phe Ile Leu Trp Asn Trp Leu 1 5 10 Tyr Leu Ile Phe Thr Ile Phe Phe Tyr Phe Ile Phe Thr Phe Ile Leu 40 Val Ala Thr Thr Ser Phe Leu. Gly 55 Lys Ile Tyr Leu Ile Val Ala Pro 25 Asn Giu. Asn Leu Thr Phe Phe Trp Val Arg Ile Phe Asn Ile Phe Leu Asp Phe Met Asn Gin Ala Tyr Phe Lys Giy Phe Leu Ile Phe His Lys 70 75 Met Lys Leu Ala Glu Lys Ile Thr Asn Phe Leu Glu Lys Thr Thr Tyr 90 Lys Lys Tyr Asn Giu. Asn Ser Ser Gly Phe Tyr Tyr Ser Glu Ile Giu 100 105 110 WO 00131115 WO 0031115PCT/AU99/01 035 120 Asn Thr Ile Giu Lys Ser Vai 115 Phe Leu Gin Thr Leu Ser Ile 130 135 Ser Gin Phe Tyr Ala 120 Asn Leu Leu Ser 125 Gly Leu Phe Phe Ile Leu Met Thr Le u 140 Tyr Ile Asn Trp Ile Leu 145 150 Ala Leu Ile Ile Val Gly Val Ile Thr 155 Phe Vai Ile Thr Thr 165 Ser Aia Lys Leu Gin 180 Ser Leu Leu Ser Lys Leu Thr Lys Leu Gin 175 Ala Ile Ser Phe Asn Asn Ser Leu Ser Thr 190 Asp Asp Lys Tyr Leu Leu Thr Leu Pro Gin Leu 195 200 Phe Giu Phe Ile Ile Asn Lys Arq 210 215 Lys Thr Leu Asn Asp Asn Lys Lys Leu Ile Ser 235 Trp Ile Thr Arg Glu 225 Lys Tyr Gly Ile Phe Ser 230 Phe Phe Asn Glu Tyr 240 Giy Ile Ala Phe Trp 255 Ser Asn Asn Phe Phe Ser Ala Ile Ile Thr Ile 245 250 Thr Leu Tyr Tyr Lys Asn Asn Asn Ser 260 265 WO 00/31115 WO 0031115PCT/AU99/01 035 <210> <211> <212> <213> <220> <223> 121 453

DNA

Artificial Sequence Description of Artificial Sequence:Clone pAD702 <400> tcttatgaga ttttttcttt tttcgttccc cttgtaaaaa tttgcaacaa aatattcaaa gttatgctaa ggtcccattt aaaaatattt ttattaaaac agttgcaaaa aagtgaatca ttatgatggg attctgccga tttatgattt atcataaaag acctttgcta acttttagac taatgcaaat aaaagaggta cttaacttac atatataact aatgcttcgt ttttgatata atagtccctg tataaagcaa cgacttgaaa aatgctatct tgagtgttat gcaatgqcag cggcgtttqg ggt gaatttttgg tcaaaaaatc tgccaccaat gacttaqcgq taaaatatta taaatggcgc qaaatattga cgctatttta tgttatttat gattccatga ctttactttg tccggagaaa tttgtttata agcaattttt 120 180 240 300 360 420 453 <210> <211> <212> <213> <220> <223> 76 151

PRT

Artificial Sequence Description of Artificial Sequence: Deduced protein sequence from clone pAD7O2 <400> 76 Ser Tyr Glu Lys Lys Tyr Leu Pro Leu Leu Ile Val Pro Gly Ile Phe 1 WO 00/31115 WO 0031115PCT/AU99/01 035 Gly Ala Ile Leu Phe Phe Leu Phe Ile Lys Thr Leu Leu 25 Ala Ile Lys Lys Ser Val Ile Tyr Phe Arg Ser Gin Leu 40 Ala Asn Arg Leu Glu Met Pro Pro Met Ile Pro Trp Leu 55 Asp Tyr Lys Gin Asn Asn Val Lys Lys Phe Thr Leu Giy Asn Gin Lys Giu Val Asn Aia Ile Trp Leu Ser 75 Phe Ala Thr Ile Met Met Gly Leu Thr Tyr Trp Val Leu Leu Lys Tyr 90 Tyr Pro Giu Ala Val Asn 115 Leu Arg Arg 130 Lys Asn Ile Gin Asn Ser Ala Glu Tyr Ile Thr Ala Met 100 105 110 Giy Ala Leu Phe Ile Val Met Leu Ile Tyr Asp Leu Met 120 125 Arg Leu Gly Asn Ile Giu Ala Ile Phe Gly Pro Ile Tyr 135 140 His Lys Ser Phe Asp Ile Giy 145 150 <210> 77 <211> 99 WO 00/31115 PCT/AU99/01 035 123 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence:Clone pAD763 <400> 77 gatctttgtg ttatttttgt taataaaacc aagtttaaaa gtcatttccc ctggtttgtc agtggtttta acatcataat aatagacaaa ataattctt 99 <210> 78 <211> 33 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence:Deduced protein sequence from pAD763 <400> 78 Asp Leu Cys Val Ile Phe Val Asn Lys Thr Lys Phe Lys Ser His Phe 1 5 10 Pro Trp Phe Val Ser Gly Phe Asn Ile Ile Ile Ile Asp Lys Ile Ile 25 Leu WO 00/31115 124 <210> 79 <211> 489 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence:Clone pAD766 <400> 79 PCT/AU99/01 035 gatcaacaaa aaaccqccga gttaatgatt tattataaaa ttagaaaagg cgaattgata cttgagaata ggcagattcg ttgaaaaga aaccacaacc tagttcaagg tttataaata atacagcaaa aaataaagaa ttggtggttt atttatcaag ataacaatta aaaagaagaa tcctagtcca caagtttaac tttttctcag acaagatccq tctaacaaaa cctttcgctt ttatataatt aaagaagaaa aaaccacaaa gataaaattc ggtggccttt gataatccta ggcacacttt tttgatagat ggatccgata aacaaqaaaa aqattgaaaa ataaatttga atagtgcaaa aaatatttta taccatttca attcccaatt aggttgagga agaagaaaaa tatcggtctt accgactgag tttgctcgaa tgttcaaaga acccgcaaat tctgaggagc atttgatagg 120 180 240 300 360 420 480 489 <210> <211> <212> <213> <220> <223> 163

PRT

Artificial Sequence Description of Artificial Sequence:Deciuced protein sequence from clone pAD766 <400> Asp Gln Gln Lys Pro Gin Pro Lys Glu Glu Lys Glu Glu Lys Gln Glu 1 5 10 WO 00/31115 PCT/AU99/01035 125 Lys Giu Glu Lys Lys Pro Pro Ile Vai Gin Gly Pro Ser Pro Lys Pro 25 Gin Lys Ile Glu Asn Ile Gly Leu Val Asn Asp Phe Tyr Lys Tyr Lys 40 Phe Asn Asp Lys Ile his Lys Phe Giu Pro Thr Glu Tyr Tyr Lys Asn 55 Thr Ala Asn Phe Ser Gin Giy Gly Leu Tyr Ser Aia Asn Leu Leu Giu 70 75 Leu Giu Lys Giu Ile Lys Lys Gin Asp Pro Asp Asn Pro Lys Ile Phe 90 Tyr Vai Gin Leu Leu Pro 115 Arg Ar Ile Asp Ile 100 Phe Gin Pro Aia Asn 120 Giy Gly Phe Leu Thr Lys Gly Thr 105 110 Leu Giu Asn Asn Leu Ser Ser Leu 125 Phe Leu Arg Ser Gly Arg Phe Asp 140 Ser Leu 130 Phe Asp Arg Tyr Ser Gin 135 Asn 145 Asn Tyr Tyr Ile Ile Giy Ser Asp Lys Val Giu Giu Phe Asp Arg 150 155 160 Leu Lys Arg WO 00131115 WO 0031115PCT/AU99/01 035 <210> <211> <212> <213> <220> <223> 126 81 474

DNA

Artificial Sequence Description of Artificial Sequence:Clone pAD957 <400> 81 agctatttta agcaaatctg gaaaaaaatt tttgtcagta gatatttttt agagttgaat cccttattgt acaaaaaata gtattataag aattatcaaa taactagatt cttttcattc tacaacaatt atgaaaataa tgagaatttc gcctatttag ccctttgttt aatagaaagt taccgcaact gctaaattca tagtgtgaaa agaaaaagat ggattctatt cttaaggttt ttggctgttt aatattttta ttagtaaaaa ataaaaaaca aatttagaaa atcatagttt gattttcagg ctcaccgatc cttgcacaaa attttataat aaactaataa atatacagca ctaaattaaa tttcgctaga ttctagagga ttaaacgaag cattataatt aaacqaaaat taacttgact acaagttttt atcaaaaatt tataaaggaa ttttacaaac att t <210> <211> <212> <213> <220> <223> 82 158

PRT

Artificial Sequence Description of Artificial Sequence:Deduced protein sequence from clone pAD957 <400> 82 Ser Tyr Phe Ser Ile Ile Ser Pro Leu Phe Leu Ala Val Ser Cys Thr WO 00/31115 WO 0031115PCT/AU99/01 035 Asn Ile Ile Ile Ser Lys Ser Giu Leu Ser Lys Ile Glu Ser Asn Ile 25 Phe Asn Phe Ile Ile Asn Giu Asn Giu Lys Asn Leu Thr Arg Phe Thr 40 Ala Thr Phe His Leu Val Lys Lys Thr Asn Asn Asn Leu Thr Phe Vai Ser Thr 55 Ser Leu Asn Ser Ile Lys Asn Asn Ile Gin Gin Gin Val Phe 70 75 Asp Ile Phe Leu Gin Gin Phe Ser Val Lys Asn Leu Glu Thr 90 Lys Leu Lys Ser Lys Val Phe Ser 115 Ser Ile Asp 130 Arg Val Giu Tyr Giu Asn Lys Glu Lys 105 Asp Ile Ile 110 Ile Ser Asp Leu Asp Ile Lys Phe Gin Vai Leu 135 Pro Leu Leu Leu Arg 125 Glu Asp Phe Thr Asn 140 Thr Lys Asn Ser Leu Phe Ser Leu Arg Phe Leu 145 150 Thr Asp Leu Lys Arg Arg Phe 155 WO 00/31115 PCT/AU99/01035 128 <210> 83 <211> 123 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence:Clone pAD996 <400> 83 agctatctat taattatgat gccgcagggg gaacctatca cactactggt taaggtcatt acgctagcgg ttatgcactc agatgaaaat gctgaaaggt acataaactc tgatgatccg 120 atc 123 <210> 84 <211> 41 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence:Deduced protein sequence from pAD996 <400> 84 Ser Tyr Leu Leu Ile Met Met Pro Gin Gly Glu Pro lie Thr Leu Leu 1 5 10 Val Lys Val lie Thr Leu Ala Val Met His Ser Asp Glu Asn Ala Glu 25 Arg Tyr Ile Asn Ser Asp Asp Pro Ile

Claims (19)

1. A method of screening to identify expression clones in a library said method comprising: providing a source of nucleotide sequences; providing an expression vector including a small marker tag; introducing a nucleotide sequence into the expression vector; transforming the expression vector into a host cell to create an expression library; and locating an expression clone by detecting the expression of a fusion protein comprising the small marker tag co-expressed with an expression protein.

2. A method of screening a library of nucleotide sequences to identify expressed nucleotide sequences, said method including the steps of: providing a source of nucleotide sequences; providing an expression vector including a small marker tag; creating a library by introducing a nucleotide sequence into the expression vector; identifying an expression protein expressed by the expression vector by determining the presence of a fusion protein comprising the small marker tag co- expressed with the expression protein translated by the nucleotide sequence; and determining the nucleotide sequence of the expression protein.

3. A method of identifying antigenic gene sequences from a sample of nucleotide sequences, said method comprising the steps of: providing an expression vector including a small marker tag; introducing a nucleotide sequence from the sample into the expression o eoeap r r vector; identifying an antigenic expression protein expressed by the expression vector by determining the presence of a fusion protein comprising the small marker tag, co-expressed with the expression protein translated by the nucleotide sequence; identifying the antigenic expression protein; 161 determining the gene sequence of the antigenic expression protein encoded by the expression vector.

4. A method of identifying a therapeutic antigenic gene sequence encoding a therapeutic antigenic protein of a disease, from a sample of nucleotide sequences, said method comprising the steps of: providing an expression vector including a small marker tag; introducing a nucleotide sequence from the sample into the expression vector; identifying an antigenic expression protein expressed by the expression vector by determining the presence of a fusion protein comprising the small marker tag co-expressed with the expression protein translated by the nucleotide sequence; inoculating an animal with the expression vector which expresses the antigenic expression protein; challenging the animal with the disease; identifying an expression vector and expression protein that provides a therapeutic effect induced by vaccination; and determining the gene sequence of the antigenic expression protein 20 encoded by the expression vector. A method according to any one of claims 1 to 4 wherein the library is a library of nucleotide sequences which translate to polypeptides of Mycoplasma, said method comprising the steps of: providing a source of nucleotide sequences from a sample of Mycoplasma; "••."providing an expression vector including a small marker tag; introducing a nucleotide sequence into the expression vector; identifying a Mycoplasma expression protein expressed by the expression vector by determining the presence of a fusion protein comprising the small S 30 marker tag co-expressed with the expression protein translated by a nucleotide sequence; and determining the nucleotide sequence of the expression protein of Mycoplasma. 162

6. A method according to claim 5 wherein the Mycoplasma is Mycoplasma hyopneumoniae.

7. A method according to any one of claims 1 to 6 wherein the small marker tag is a polyHis tag.

8. A nucleotide sequence of Mycoplasma having a sequence according to any one of Figures 1 to 42, or SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:41, SEQ ID NO:43, SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53, SEQ ID NO:55, SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:61, SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:67, SEQ ID NO:69, SEQ ID NO:71, SEQ ID NO:73, SEQ ID NO:75, SEQ ID NO:77, SEQ ID NO:79, SEQ ID NO:81 or SEQ ID NO:83 or mutants, analogues, derivatives or functionally active fragments thereof. 20 9. A recombinant vector including a nucleotide sequence according to claim 8. A host cell including a recombinant vector according to claim 9.

11. An amino acid sequence of Mycoplasma having a sequence according to any one of Figures 1 to 42, or SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ .ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:0, SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:0, SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:58, SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:64, SEQ ID NO:66, SEQ ID NO:68, SEQ ID NO:70, SEQ ID NO:72, SEQ ID NO:74, SEQ ID NO:76, SEQ W:\Fiesl642861\642801Spei.doc 163 ID NO:78, SEQ ID NO:80, SEQ ID NO:82 or SEQ ID NO:84 or a mutant, analogue, derivative or functionally active fragment thereof.

12. An isolated and purified polypeptide encoded by an amino acid sequence according to claim 11.

13. An isolated and purified polypeptide translated from a nucleotide sequence according to claim 8.

14. An antibody specific for a polypeptide according to claim 12. An antibody specific for a polypeptide according to claim 13.

16. An antibody according to claim 14 or 15 which is a monoclonal antibody.

17. A method of treating an infection, said method including administering an effective amount of an antigenic polypeptide according to claim 12 or 13.

18. A method of treating an infection, said method including administering to an animal in need, an effective amount of a DNA molecule including a nucleotide sequence according to claim 8.

19. A method according to claim 18 wherein the DNA molecule is administered via a recombinant vector according to claim 9. A method of treating an infection, said method including administering an effective amount of an antibody specific to a polypeptide, according to any one of claims 14 to 16.

21. A method according to any one of claims 17 to 20 wherein the infection is a Mycoplasma infection.

22. A method according to claim 21 wherein the infection is a Mycoplasma hyopneumoniae infection. W:'Files\642861\642861_Sped.doc 164

23. A method according to any one of claims 18, 19, 21 or 22 wherein the nucleotide sequence comprises a sequence according to any one of Figures 1 to 42, or SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:41, SEQ ID NO:43, SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53, SEQ ID NO:55, SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:61, SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:67, SEQ ID NO:69, SEQ ID NO:71, SEQ ID NO:73, SEQ ID NO:75, SEQ ID NO:77, SEQ ID NO:79, SEQ ID NO:81 or SEQ ID NO:83 or a mutant, analogue, derivative or functionally active fragment thereof.

24. A method according to any one of claims 17, 20 to 22 wherein the polypeptide is encoded by an amino acid sequence according to any one of Figures 1 to 42, SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO:40, SEQ ID NO:42, SEQ ID NO:44, SEQ ID NO:46, SEQ ID NO:48, SEQ ID NO:50, SEQ ID NO:52, SEQ ID NO:54, SEQ ID NO:56, SEQ ID NO:58, SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:64, SEQ ID NO:66, SEQ ID NO:68, SEQ ID NO:70, SEQ ID NO:72, SEQ ID NO:74, SEQ ID NO:76, SEQ ID NO:78, SEQ ID NO:80, SEQ ID NO:82 or SEQ ID NO:84 or a mutant, analogue, derivative or functionally active fragment thereof. Dated: 10June2003 30 PHILLIPS ORMONDE FITZPATRICK Attorneys for: COMMONWEALTH SCIENTIFIC and INDUSTRIAL RESEARCH ORGANISATION W:\Files\642861\64286_1Sped.doc