AU733388B2 - Materials and methods for the modification of plant lignin content - Google Patents

Description

WO 98/11205 PCT/NZ97/00112 MATERIALS AND METHODS FOR THE MODIFICATION OF PLANT LIGNIN CONTENT Technical Field of the Invention This invention relates to the field of modification of lignin content and composition in plants. More particularly, this invention relates to enzymes involved in the lignin biosynthetic pathway and nucleotide sequences encoding such enzymes.

Backaround of the Invention Lignin is an insoluble polymer which is primarily responsible for the rigidity of plant stems. Specifically, lignin serves as a matrix around the polysaccharide components of some plant cell wails. The higher the lignin content, the more rigid the plant. For example, tree species synthesize large quantities of lignin, with lignin constituting between 20% to 30% of the dry weight of wood. In addition to providing rigidity, lignin aids in water transport within plants by rendering cell walls hydrophobic and water impermeable. Lignin also plays a role in disease resistance of plants by impeding the penetration and propagation of pathogenic agents.

The high concentration of lignin in trees presents a significant problem in the paper industry wherein considerable resources must be employed to separate lignin from the cellulose fiber needed for the production of paper. Methods typically employed for the removal of lignin are highly energy- and chemical-intensive, resulting in increased costs and increased levels of undesirable waste products. In the U.S. alone, about 20 million tons of lignin are removed from wood per year.

Lignin is largely responsible for the digestibility, or lack thereof, of forage crops, with small increases in plant lignin content resulting in relatively high decreases in digestibility. For example, crops with reduced lignin content provide more efficient forage for cattle, with the yield of milk and meat being higher relative to the amount of forage crop consumed. During normal plant growth, the increase in dry matter content is accompanied by a corresponding decrease in digestibility. When deciding on the optimum time to harvest forage crops, farmers must therefore chose between a high yield of less digestible material and a lower yield of more digestible material.

f, 6 WO 98/11205 PCT/NZ97/00112 For some applications, an increase in lignin content is desirable since increasing the lignin content of a plant would lead to increased mechanical strength of wood, changes in its color and increased resistance to rot. Mycorrhizal species composition and abundance may also be favorably manipulated by modifying lignin content and structural composition.

As discussed in detail below, lignin is formed by polymerization of at least three different monolignols which are synthesized in a multistep pathway, each step in the pathway being catalyzed by a different enzyme. It has been shown that manipulation of the number of copies of genes encoding certain enzymes, such as cinnamvl alcohol dehydrogenase (CAD) and caffeic acid 3-0-methyltransferase (COMT) results in modification of the amount of lignin produced; see, for example, U.S. Patent No.

5,451,514 and PCT publication no. WO 94/23044. Furthermore, it has been shown that antisense expression of sequences encoding CAD in poplar leads to the production of lignin having a modified composition (Grand, C. et al. Planta (Berl.) 163:232-237 (1985)).

While DNA sequences encoding some of the enzymes involved in the lignin biosynthetic pathway have been isolated for certain species of plants, genes encoding many of the enzymes in a wide range of plant species have not yet been identified.

Thus there remains a need in the art for materials useful in the modification of lignin content and composition in plants and for methods for their use.

Summary of the Invention Briefly, the present invention provides isolated DNA sequences obtainable from eucalyptus and pine which encode enzymes involved in the lignin biosynthetic pathway, DNA constructs including such sequences, and methods for the use of such constructs. Transgenic plants having altered lignin content and composition are also provided.

In a first aspect, the present invention provides isolated DNA sequences coding for the following enzymes isolated from eucalyptus and pine: cinnamate 4-hydroxylase (C4H), coumarate 3-hydroxylase (C3H), phenolase (PNL), O-methyl transferase (OMT), cinnamyl alcohol dehydrogenase (CAD), cinnamoyl-CoA reductase (CCR), phenylalanine ammonia-lyase (PAL), 4-coumarate:CoA ligase (4CL), coniferol glucosyl transferase (CGT), coniferin beta-glucosidase (CBG), laccase (LAC) and peroxidase (POX), together with ferulate-5-hydroxylase (F5H) from eucalyptus. In one embodiment, the isolated DNA sequences comprise a nucleotide sequence selected from the group consisting of: sequences recited in SEQ ID NO: 3, 13, 16-70, and 72-88; complements of the sequences recited in SEQ ID NO: 3, 13, 16-70, 72-88; reverse complements of the sequences recited in SEQ ID NO: 3, 13, 16-70, 72-88; (d) reverse sequences of the sequences recited in SEQ ID NO: 3, 13, 16-70, 72- 88; and sequences having at least about a 99% probability of being the to same as a sequence of as measured by the computer algorithm FASTA. In a preferred embodiment, the isolated DNA sequences comprise a nucleotide sequence selected from the group consisting of: nucleotides 1-535 of SEQ ID NO: 1; nucleotides 46-671 of SEQ ID NO: 2; nucleotides 1-535 of

*SSS

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15 SEQ

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20 SEQ

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ID NO: ID NO: ID NO: ID NO: ID NO: ID NO: ID NO: ID NO: ID NO: ID NO: ID NO: ID NO: ID NO: nucleotides 290-949 of SEQ ID NO: 4; nucleotides 15-959 nucleotides 15-1026 of SEQ ID NO: 6; nucleotides 15-1454 nucleotides 15-740 of SEQ ID NO: 8; nucleotides 108-624 9; nucleotides nucleotides nucleotides nucleotides nucleotides nucleotides nucleotides nucleotides nucleotides nucleotides 68-274 of SEQ ID NO: 10; nucleotides 1-765 1-384 of SEQ ID NO: 12; nucleotides 1-278 14-472 of SEQ ID NO: 16; nucleotides 1-672 15-469 of SEQ ID NO: 18; nucleotides 15-469 1-341 of SEQ ID NO: 20; nucleotides 15-387 1-443 of SEQ ID NO: 22; nucleotides 15-607 15-421 of SEQ ID NO: 24; nucleotides 1-760 58-469 of SEQ ID NO: 26; nucleotides 15-495 15-472 of SEQ ID NO: 28; nucleotides 13-396 15-592 of SEQ ID NO: 30; nucleotides 15-468 SEQ ID NO: 31; nucleotides 1-405 of SEQ ID NO: 32; nucleotides 1-380 of 7/03/01 3a SEQ ID NO: 33; nucleotides 1-305 of SEQ ID NO: 34; nucleotides 15-693 of SEQ ID NO: 35; nucleotides 1-418 of SEQ ID NO: 36; nucleotides 15-777 of SEQ ID NO: 37; nucleotides 1-344 of SEQ ID NO: 38; nucleotides 1-341 of SEQ ID NO: 39; nucleotides 15-358 of SEQ ID NO: 40; nucleotides 1-409 of SEQ ID NO: 41; nucleotides 1-515 of SEQ ID NO: 42; nucleotides 15-471 of SEQ ID NO: 43; nucleotides 15-487 of SEQ ID NO: 44; nucleotides 108-664 of SEQ ID NO: 45; nucleotides 15-418 of SEQ ID NO: 46; nucleotides 65-479 of SEQ ID NO: 47; nucleotides 127-1785 of SEQ ID NO: 48; nucleotides 15-475 of SEQ ID NO: 49; nucleotides 288-801 of SEQ ID NO: 50; nucleotides 51- 711 of SEQ ID NO: 51; nucleotides 1-426 of SEQ ID NO: 52; nucleotides 92- 562 of SEQ ID NO: 53; nucleotides 1-1074 of SEQ ID NO: 54; nucleotides 1- 1075 of SEQ ID NO: 55; nucleotides 1-1961 of SEQ ID NO: 56; nucleotides 1- 1010 of SEQ ID NO: 57; nucleotides 15-741 of SEQ ID NO: 58; nucleotides 1- 643 of SEQ ID NO: 59; nucleotides 15-441 of SEQ ID NO: 60; nucleotides 15 913 of SEQ ID NO: 61; nucleotides 15-680 of SEQ ID NO: 62; nucleotides 492 of SEQ ID NO: 63; nucleotides 15-524 of SEQ ID NO: 64; nucleotides 1- S: 492 of SEQ ID NO: 63; nucleotides 1-524 of SEQ ID NO: 64; nucleotides 176- :609417 of SEQ ID NO: 65; nucleotides 1-511 of SEQ ID NO: 66; nucleotides 176- 609 of SEQ ID NO: 67; nucleotides 1-4746-608 of SEQ ID NO: 68; nucleotides 1-474 1474 of SEQ ID NO: 71 nucleotides 15-1038 of SEQ ID NO: 7; nucleotides 1- 372 of SEQ ID NO: 73; nucleotides 18-545 of SEQ ID NO: 74; nucleotides 4637 of SEQ ID NO: 75; nucleotides 32-435 of SEQ ID NO: 76; nucleotides 3451 of SEQ ID NO: 77; nucleotides 1-374 of SEQ ID NO: 78; nucleotides 1-457 of SEQ ID NO: 79; nucleotides 1-346 of SEQ ID NO: 80; nucleotides 15-957 of SEQ ID NO: 81; nucleotides 40-452 of SEQ ID NO: 82; nucleotides 15-471 of SEQ ID NO: 83; nucleotides 1-338 of SEQ ID NO: 84; nucleotides 150-1229 of 7/03/01 SEQ ID NO: 85; nucleotides 1-1410 of SEQ ID NO: 86; nucleotides 1-687 of SEQ ID NO: 87; and nucleotides 1-688 of SEQ ID NO: 88.

In another aspect, the invention provides DNA constructs comprising a DNA sequence of the present invention, either alone, in combination with one or more of the inventive sequences or in combination with one or more known DNA sequences; together with transgenic cells comprising such constructs.

In a related aspect, the present invention provides DNA constructs comprising, in the direction, a gene promoter sequence; an open reading frame coding for at least a functional portion of an enzyme encoded by the io inventive DNA sequences or variants thereof; and a gene termination sequence. The open reading frame may be orientated in either a sense or antisense direction. DNA constructs comprising a non-coding region of a gene coding for an enzyme encoded by the above DNA sequences or a nucleotide sequence complementary to a non-coding region, together with a gene :5 promoter sequence and a gene termination sequence, are also provided.

Preferably, the gene promoter and termination sequences are functional in a host plant. Most preferably, the gene promoter and termination sequences are those of the original enzyme genes but others generally used in the art, such as the Cauliflower Mosaic Virus (CMV) promoter, with or without enhancers, such as the Kozak sequence or Omega enhancer, and Agrobacterium tumefaciens nopalin synthase terminator may be usefully employed in the present invention. Tissue-specific promoters may be employed in order to target expression to one or more desired tissues. In a preferred embodiment, the gene promoter sequence provides for transcription in xylem. The DNA construct may further include a marker for the identification of transformed cells.

7/03/01 WO 98/11205 PCT/NZ97/00112 In a further aspect, transgenic plant cells comprising the DNA constructs of the present invention are provided, together with plants comprising such transgenic cells, and fruits and seeds of such plants.

In yet another aspect, methods for modulating the lignin content and composition of a plant are provided, such methods including stably incorporating into the genome of the plant a DNA construct of the present invention. In a preferred embodiment, the target plant is a woody plant, preferably selected from the group consisting of eucalyptus and pine species, most preferably from the group consisting of Eucalyptus grandis and Pinus radiata. In a related aspect, a method for producing a plant having altered lignin content is provided, the method comprising transforming a plant cell with a DNA construct of the present invention to provide a transgenic cell, and cultivating the transgenic cell under conditions conducive to regeneration and mature plant growth.

In yet a further aspect, the present invention provides methods for modifying the activity of an enzyme in a plant, comprising stably incorporating into the genome of the plant a DNA construct of the present invention. In a preferred embodiment, the target plant is a woody plant, preferably selected from the group consisting of eucalyptus and pine species, most preferably from the group consisting of Eucalyptus grandis and Pinus radiata.

The above-mentioned and additional features of the present invention and the manner of obtaining them will become apparent, and the invention will be best understood by reference to the following more detailed description, read in conjunction with the accompanying drawing.

Brief Description of the Figures Fig. 1 is a schematic overview of the lignin biosynthetic pathway.

Detailed Description Lignin is formed by polymerization of at least three different monolignols, primarily para-coumaryl alcohol, coniferyl alcohol and sinapyl alcohol. While these three types of lignin subunits are well known, it is possible that slightly different variants of these subunits may be involved in the lignin biosynthetic pathway in various WO 98/11205 PCT/NZ97/00112 plants. The relative concentration of these residues in lignin varies between different plant species and within species. In addition, the composition of lignin may also vary between different tissues within a specific plant. The three monolignols are derived from phenylalanine in a multistep process and are believed to be polymerized into lignin by a free radical mechanism.

Fig. 1 shows the different steps in the biosynthetic pathway for coniferyl alcohol together with the enzymes responsible for catalyzing each step. para-Coumaryl alcohol and sinapyl alcohol are synthesized by similar pathways. Phenylalanine is first deaminated by phenylalanine ammonia-lyase (PAL) to give cinnamate which is then hydroxylated by cinnamate 4-hydroxylase (C4H) to form p-coumarate. p-Coumarate is hydroxylated by coumarate 3-hydroxylase to give caffeate. The newly added hydroxyl group is then methylated by O-methyl transferase (OMT) to give ferulate which is conjugated to coenzyme A by 4-coumarate:CoA ligase (4CL) to form feruloyl-CoA.

Reduction of feruloyl-CoA to coniferaldehyde is catalyzed by cinnamoyl-CoA reductase (CCR). Coniferaldehyde is further reduced by the action of cinnamyl alcohol dehydrogenase (CAD) to give coniferyl alcohol which is then converted into its glucosylated form for export from the cytoplasm to the cell wall by coniferol glucosyl transferase (CGT). Following export, the de-glucosylated form of coniferyl alcohol is obtained by the action of coniferin beta-glucosidase (CBG). Finally, polymerization of the three monolignols to provide lignin is catalyzed by phenolase (PNL), laccase (LAC) and peroxidase (POX).

The formation of sinapyl alcohol involves an additional enzyme, hydroxylase (F5H). For a more detailed review of the lignin biosynthetic pathway, see: Whetton, R. and Sederoff, The Plant Cell, 7:1001-1013 (1995).

Quantitative and qualitative modifications in plant lignin content are known to be induced by external factors such as light stimulation, low calcium levels and mechanical stress. Synthesis of new types of lignins, sometimes in tissues not normally lignified, can also be induced by infection with pathogens. In addition to lignin, several other classes of plant products are derived from phenylalanine, including flavonoids, coumarins, stilbenes and benzoic acid derivatives, with the initial steps in the synthesis of all these compounds being the same. Thus modification of the action of PAL, C4H and 4CL may affect the synthesis of other plant products in addition to lignin.

6 Using the methods and materials of the present invention, the lignin content of a plant can be increased by incorporating additional copies of genes encoding enzymes involved in the lignin biosynthetic pathway into the genome of the target plant. Similarly, a decrease in lignin content can be obtained by transforming the target plant with antisense copies of such genes. In addition, the number of copies of genes encoding for different enzymes in the lignin biosynthetic pathway can be manipulated to modify the relative amount of each monolignol synthesized, thereby leading to the formation of lignin having altered composition. The alteration of lignin composition would be advantageous, for example, in tree processing for paper, and may also be effective in altering the palatability of wood materials to rotting fungi.

In one embodiment, the present invention provides isolated complete or partial DNA sequences encoding, or partially encoding, enzymes involved 15 in the lignin biosynthetic pathway, the DNA sequences being obtainable from eucalyptus and pine. Specifically, the present invention provides isolated DNA sequences encoding the enzymes CAD (SEQ ID NO: 1, in 96*.

particular nucleotides 1 535; and SEQ ID NO: 30, in particular nucleotides 15 592), PAL (SEQ ID NO: 16, in particular nucleotides 14 20 472), C4H (SEQ ID NO: 17, in particular nucleotides 1 672), C3H (SEQ ID NO: 18, in particular nucleotides 15 469), F5H (SEQ ID NO: 19, in particular nucleotides 15 469; SEQ ID NO: 20, in particular nucleotides 1 341; and SEQ ID NO: 21, in particular nucleotides 15 387), OMT (SEQ ID NO: 22, in particular nucleotides 1 443; SEQ ID NO: 23, in particular nucleotides 15 607; SEQ ID NO: 24, in particular nucleotides 15 421; and SEQ ID NO: 25, in particular nucleotides 1 760), CCR (SEQ ID NO: 26, in particular nucleotides 58 469; SEQ ID NO: 27, in particular 7/03/01 nucleotides 15 495; SEQ ID NO: 28, in particular nucleotides 15 472; and SEQ ID NO: 29, in particular nucleotides 13 396), CGT (SEQ ID NO: 31, in particular nucleotides 15 468; SEQ ID NO: 32, in particular nucleotides 1 405; and SEQ ID NO: 33, in particular nucleotides 1 380), CBG (SEQ ID NO: 34, in particular nucleotides 1 305), PNL (SEQ ID NO: in particular nucleotides 15 693; and SEQ ID NO: 36, in particular nucleotides 1 418), LAC (SEQ ID NO: 37, in particular nucleotides 15 777; SEQ ID NO: 38, in particular nucleotides 1 344; SEQ ID NO: 39, in particular nucleotides 1 341; SEQ ID NO: 40, in particular nucleotides 358; and SEQ ID NO: 41, in particular nucleotides 1 409) and POX (SEQ ID NO: 42, in particular nucleotides 1 515; SEQ ID NO: 43, in particular nucleotides 15 571; and SEQ ID NO: 44, in particular nucleotides 15 487) from Eucalyptus grandis; and the enzymes C4H (SEQ ID NO: 2, in particular nucleotides 46 671; SEQ ID NO: 3, in particular S 15 nucleotides 1 535; SEQ ID NO: 48, in particular nucleotides 127 1785; and SEQ ID NO: 49, in particular nucleotides 15 475), C3H (SEQ ID NO: 4, in particular nucleotides 290 949; SEQ ID NO: 50, in particular nucleotides 288 801; SEQ ID NO: 51, in particular nucleotides 51 711; and SEQ ID NO: 52, in particular nucleotides 1 426), PNL (SEQ ID NO: 20 in particular nucleotides 15 959; and SEQ ID NO: 81, in particular nucleotides 15 957), OMT (SEQ ID NO: 6, in particular nucleotides 15 1026; SEQ ID NO: 53, in particular nucleotides 92 562; SEQ ID NO: 54, eat* in particular nucleotides 1 1074; and SEQ ID NO: 55, in particular nucleotides 1 1075), CAD (SEQ ID NO: 7, in particular nucleotides 15 1454; and SEQ ID NO: 71, in particular nucleotides 15 1474), CCR (SEQ ID NO: 8, in particular nucleotides 15 740; SEQ ID NO: 58, in particular nucleotides 15 741; SEQ ID NO: 59, in particular nucleotides 1 643; 7/03/01 6b SEQ ID NO: 60, in particular nucleotides 15 441; SEQ ID NO: 61, in particular nucleotides 15 913; SEQ ID NO: 62, in particular nucleotides 680; SEQ ID NO: 63, in particular nucleotides 15 492; SEQ ID NO: 64, in particular nucleotides 15 524; SEQ ID NO: 65, in particular nucleotides 1 417; SEQ ID NO: 66, in particular nucleotides 1 511; SEQ ID NO: 67, in particular nucleotides 176 609; SEQ ID NO: 68, in particular nucleotides 1 474; SEQ ID NO: 69, in particular nucleotides 1 474; and SEQ ID NO: 70, in particular nucleotides 176 608), PAL (SEQ ID NO: 9, in particular nucleotides 108 624; SEQ ID NO: 10, in particular o0 nucleotides 68 274; SEQ ID NO: 11, in particular nucleotides 1 765; SEQ ID NO: 45, in particular nucleotides 108 664; SEQ ID NO: 46, in particular nucleotides 15 418; and SEQ ID NO: 47, in particular nucleotides 65 479), 4CL (SEQ ID NO: 12, in particular nucleotides 1 384; SEQ ID NO: 56, in particular nucleotides 1 1961; and SEQ ID NO: 15 57, in particular nucleotides 1 1010), CGT (SEQ ID NO: 72, in particular nucleotides 15 1038), CBG (SEQ ID NO: 73, in particular nucleotides 1 372; SEQ ID NO: 74, in particular nucleotides 18 545; SEQ ID NO: in particular nucleotides 40 463; SEQ ID NO: 76, in particular nucleotides 32 435; SEQ ID NO: 77, in particular nucleotides 15 451; SEQ ID NO: 20 78, in particular nucleotides 1 374; SEQ ID NO: 79, in particular nucleotides 1 457; and SEQ ID NO: 80, in particular nucleotides 1 346), LAC (SEQ ID NO: 82, in particular nucleotides 40 452; SEQ ID NO: 83, in particular nucleotides 15 471; and SEQ ID NO: 84, in particular nucleotides 1 338) and POX (SEQ ID NO: 13, in particular nucleotides 1 278; SEQ ID NO: 85, in particular nucleotides 150 1229; SEQ ID NO: 86, in particular nucleotides 1 1410; SEQ ID NO: 87, in particular nucleotides 1 687; and SEQ ID NO: 88, in particular nucleotides 1 688) 7/03/01 from Pinus radiata. Complements of such isolated DNA sequences, reverse complements of such isolated DNA sequences and reverse sequences of such isolated DNA sequences, together with variants of such sequences, are also provided. DNA sequences encompassed by the present invention include cDNA, genomic DNA, recombinant DNA and wholly or partially chemically synthesized DNA molecules.

The definition of the terms "complement", "reverse complement" and "reverse sequence", as used herein, is best illustrated by the following example. For the sequence 5' AGGACC the complement, reverse complement and reverse sequence are as follows: ee 7/03/01 WO 98/11205 PCT/NZ97/00112 complement 3' TCCTGG reverse complement 3' GGTCCT reverse sequence 5' CCAGGA 3'.

As used herein, the term "variant" covers any sequence which exhibits at least about 50%, more preferably at least about 70% and, more preferably yet, at least about 90% identity to a sequence of the present invention. Most preferably, a "variant" is any sequence which has at least about a 99% probability of being the same as the inventive sequence. The probability for DNA sequences is measured by the computer algorithm FASTA (version 2.0u4, February 1996; Pearson W. R. et al..

Proc. Natl. Acad. Sci., 85:2444-2448, 1988), the probability for translated DNA sequences is measured by the computer algorithm TBLASTX and that for protein sequences is measured by the computer algorithm BLASTP (Altschul, S. F. et al. J.

Mol. Biol., 215:403-410, 1990). The term "variants" thus encompasses sequences wherein the probability of finding a match by chance (smallest sum probability) in a database, is less than about 1% as measured by any of the above tests.

Variants of the isolated sequences from other eucalyptus and pine species, as well as from other commercially important species utilized by the lumber industry, are contemplated. These include the following gymnosperms, by way of example: loblolly pine Pinus taeda, slash pine Pinus elliotti, sand pine Pinus clausa, longleaf pine Pinus palustrus, shortleaf pine Pinus echinata, ponderosa pine Pinus ponderosa, Jeffrey pine Pinus jeffrey, red pine Pinus resinosa, pitch pine Pinus rigida, jack pine Pinus banksiana, pond pine Pinus serotina, Eastern white pine Pinus strobus, Western white pine Pinus monticola, sugar pine Pinus lambertiana, Virginia pine Pinus virginiana, lodgepole pine Pinus contorta, Caribbean pine Pinus caribaea, P. pinaster, Calabrian pine P. brutia, Afghan pine P. eldarica, Coulter pine P. coulteri, European pine P.

nigra and P. sylvestris; Douglas-fir Pseudotsuga menziesii; the hemlocks which include Western hemlock Tsuga heterophylla, Eastern hemlock Tsuga canadensis, Mountain hemlock Tsuga mertensiana; the spruces which include the Norway spruce Picea abies, red spruce Picea rubens, white spruce Picea glauca, black spruce Picea mariana, Sitka spruce Picea sitchensis, Englemann spruce Picea engelmanni, and blue spruce Picea pungens; redwood Sequoia sempervirens; the true firs include the Alpine fir Abies lasiocarpa, silver fir Abies amabilis, grand fir Abies grandis, noble fir Abies procera, white fir Abies concolor, California red fir Abies magnifica, and balsam fir Abies balsamea, the cedars which include the Western red cedar Thuja plicata, incense -7- WO 98/11205 PCT/NZ97/00112 cedar libocedrus decurrens, Northern white cedar Thuja occidentalis, Port Orford cedar Chamaecyparis lawsoniona, Atlantic white cedar Chamaecyparis thyoides, Alaska yellow-cedar Chamaecyparis nootkatensis, and Eastern red cedar Huniperus virginiana; the larches which include Eastern larch Larix laricina, Western larch Larix occidentalis, European larch Larix decidua, Japanese larch Larix leptolepis, and Siberian larch Larix siberica; bold cypress Taxodium distichum and Giant sequoia Sequoia gigantea; and the following angiosperms, by way of example: Eucalyptus alba, E. bancroftii, E. botyroides, E. bridgesiana, E. calophylla, E.

camaldulensis. E. citriodora, E. cladocalyx, E. coccifera, E. curtisii, E. dalrympleana, E.

deglupta E. delagatensis, E. diversicolor, E. dunnii, E. ficifolia, E. globulus, E.

gomphocephala E. gunnii, E. henryi, E. laevopinea, E. macarthurii, E. macrorhyncha.

E. maculata, E. marginata, E. megacarpa, E. melliodora, E. nicholii, E. nitens, E. novaanglica. E. obliqua, E. obtusiflora, E. oreades, E. pauciflora, E. polybractea, E. regnans, E. resinifera, E. robusta, E. rudis, E. saligna, E. sideroxylon, E. stuartiana, E. tereticoris, E. torelliana, E. umigera, E. urophylla, E. viminalis, E. viridis, E. wandoo and E.

youmanni.

The inventive DNA sequences may be isolated by high throughput sequencing of cDNA libraries such as those prepared from Eucalyptus grandis and Pinus radiata as described below in Examples 1 and 2. Alternatively, oligonucleotide probes based on the sequences provided in SEQ ID NO: 1-13 and 16-88 can be synthesized and used to identify positive clones in either cDNA or genomic DNA libraries from Eucalyptus grandis and Pinus radiata, or from other gymnosperms and angiosperms including those identified above, by means of hybridization or PCR techniques.

Probes can be shorter than the sequences provided herein but should be at least about 10, preferably at least about 15 and most preferably at least about nucleotides in length. Hybridization and PCR techniques suitable for use with such oligonucleotide probes are well known in the art. Positive clones may be analyzed by restriction enzyme digestion, DNA sequencing or the like.

In addition, the DNA sequences of the present invention may be generated by synthetic means using techniques well known in the art. Equipment for automated synthesis of oligonucleotides is commercially available from suppliers such as Perkin Elmer/Applied Biosystems Division (Foster City, CA) and may be operated according to the manufacturer's instructions.

-8- WO 98/11205 PCT/NZ97/00112 In one embodiment, the DNA constructs of the present invention include an open reading frame coding for at least a functional portion of an enzyme encoded by a nucleotide sequence of the present invention or a variant thereof. As used herein, the "functional portion" of an enzyme is that portion which contains the active site essential for affecting the metabolic step, i.e. the portion of the molecule that is capable of binding one or more reactants or is capable of improving or regulating the rate of reaction. The active site may be made up of separate portions present on one or more polypeptide chains and will generally exhibit high substrate specificity. The term "enzyme encoded by a nucleotide sequence" as used herein, includes enzymes encoded i0 by a nucleotide sequence which includes the partial isolated DNA sequences of the present invention.

For applications where amplification of lignin synthesis is desired, the open reading frame is inserted in the DNA construct in a sense orientation, such that transformation of a target plant with the DNA construct will lead to an increase in the number of copies of the gene and therefore an increase in the amount of enzyme. When down-regulation of lignin synthesis is desired, the open reading frame is inserted in the DNA construct in an antisense orientation, such that the RNA produced by transcription of the DNA sequence is complementary to the endogenous mRNA sequence. This, in turn, will result in a decrease in the number of copies of the gene and therefore a decrease in the amount of enzyme. Alternatively, regulation can be achieved by inserting appropriate sequences or subsequences DNA or RNA) in ribozyme constructs.

In a second embodiment, the inventive DNA constructs comprise a nucleotide sequence including a non-coding region of a gene coding for an enzyme encoded by a DNA sequence of the present invention, or a nucleotide sequence complementary to such a non-coding region. As used herein the term "non-coding region" includes both transcribed sequences which are not translated, and non-transcribed sequences within about 2000 base pairs 5' or 3' of the translated sequences or open reading frames.

Examples of non-coding regions which may be usefully employed in the inventive constructs include introns and 5'-non-coding leader sequences. Transformation of a target plant with such a DNA construct may lead to a reduction in the amount of lignin synthesized by the plant by the process of cosuppression, in a manner similar to that WO 98/11205 PCT/NZ97/00112 discussed, for example, by Napoli et al. (Plant Cell 2:279-290, 1990) and de Carvalho Niebel et al. (Plant Cell 7:347-358, 1995).

The DNA constructs of the present invention further comprise a gene promoter sequence and a gene termination sequence, operably linked to the DNA sequence to be transcribed, which control expression of the gene. The gene promoter sequence is generally positioned at the 5' end of the DNA sequence to be transcribed, and is employed to initiate transcription of the DNA sequence. Gene promoter sequences are generally found in the 5' non-coding region of a gene but they may exist in introns (Luehrsen, K. Mol. Gen. Genet. 225:81-93, 1991) or in the coding region, as for l0 example in PAL of tomato (Bloksberg, 1991. Studies on the Biology of Phenylalanine Ammonia Lyase and Plant Pathogen Interaction. Ph.D. Thesis. Univ. of California, Davis, University Microfilms International order number 9217564). When the construct includes an open reading frame in a sense orientation, the gene promoter sequence also initiates translation of the open reading frame. For DNA constructs comprising either an open reading frame in an antisense orientation or a non-coding region, the gene promoter sequence consists only of a transcription initiation site having a RNA polymerase binding site.

A variety of gene promoter sequences which may be usefully employed in the DNA constructs of the present invention are well known in the art. The promoter gene sequence, and also the gene termination sequence, may be endogenous to the target plant host or may be exogenous, provided the promoter is functional in the target host.

For example, the promoter and termination sequences may be from other plant species, plant viruses, bacterial plasmids and the like. Preferably, gene promoter and termination sequences are from the inventive sequences themselves.

Factors influencing the choice of promoter include the desired tissue specificity of the construct, and the timing of transcription and translation. For example, constitutive promoters, such as the 35S Cauliflower Mosaic Virus (CaMV promoter, will affect the activity of the enzyme in all parts of the plant. Use of a tissue specific promoter will result in production of the desired sense or antisense RNA only in the tissue of interest. With DNA constructs employing inducible gene promoter sequences, the rate of RNA polymerase binding and initiation can be modulated by external stimuli, such as light, heat, anaerobic stress, alteration in nutrient conditions WO 98/11205 PCT/NZ97/00112 and the like. Temporally regulated promoters can be employed to effect modulation of the rate of RNA polymerase binding and initiation at a specific time during development of a transformed cell. Preferably, the original promoters from the enzyme gene in question, or promoters from a specific tissue-targeted gene in the organism to be transformed, such as eucalyptus or pine are used. Other examples of gene promoters which may be usefully employed in the present invention include, mannopine synthase (mas), octopine synthase (ocs) and those reviewed by Chua et al. (Science. 244:174- 181, 1989).

The gene termination sequence, which is located 3' to the DNA sequence to be transcribed, may come from the same gene as the gene promoter sequence or may be from a different gene. Many gene termination sequences known in the art may be usefully employed in the present invention, such as the 3' end of the Agrobacterium tumefaciens nopaline synthase gene. However, preferred gene terminator sequences are those from the original enzyme gene or from the target species to be transformed.

The DNA constructs of the present invention may also contain a selection marker that is effective in plant cells, to allow for the detection of transformed cells containing the inventive construct. Such markers, which are well known in the art, typically confer resistance to one or more toxins. One example of such a marker is the NPTII gene whose expression results in resistance to kanamycin or hygromycin, antibiotics which is usually toxic to plant cells at a moderate concentration (Rogers et al. in Methods for Plant Molecular Biologv, A. Weissbach and H. Weissbach. eds., Academic Press Inc., San Diego, CA (1988)). Alternatively, the presence of the desired construct in transformed cells can be determined by means of other techniques well known in the art, such as Southern and Western blots.

Techniques for operatively linking the components of the inventive DNA constructs are well known in the art and include the use of synthetic linkers containing one or more restriction endonuclease sites as described, for example, by Maniatis et al., (Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratories, Cold Spring Harbor, NY, 1989). The DNA construct of the present invention may be linked to a vector having at least one replication system, for example, E. coli, whereby after each manipulation, the resulting construct can be cloned and sequenced and the correctness of the manipulation determined.

WO 98/11205 PCT/NZ97/00112 The DNA constructs of the present invention may be used to transform a variety of plants, both monocotyledonous grasses, corn, grains, oat, wheat and barley), dicotyledonous Arabidopsis, tobacco, legumes, alfalfa, oaks, eucalyptus, maple), and Gymnosperms Scots pine (Aronen, Finnish Forest Res. Papers, vol. 595, 1996), white spruce (Ellis et al., Biotechnologv 11:94-92, 1993), larch (Huang et al., In Vitro Cell 27:201-207, 1991). In a preferred embodiment, the inventive DNA constructs are employed to transform woody plants, herein defined as a tree or shrub whose stem lives for a number of years and increases in diameter each year by the addition of woody tissue. Preferably the target plant is selected from the group consisting of eucalyptus and pine species, most preferably from the group consisting of Eucalyptus grandis and Pinus radiata. As discussed above, transformation of a plant with a DNA construct including an open reading frame coding for an enzyme encoded by an inventive DNA sequence wherein the open reading frame is orientated in a sense direction will lead to an increase in lignin content of the plant or, in some cases, to a decrease by cosuppression. Transformation of a plant with a DNA construct comprising an open reading frame in an antisense orientation or a non-coding (untranslated) region of a gene will lead to a decrease in the lignin content of the transformed plant.

Techniques for stably incorporating DNA constructs into the genome of target plants are well known in the art and include Agrobacterium tumefaciens mediated introduction, electroporation, protoplast fusion, injection into reproductive organs, injection into immature embryos, high velocity projectile introduction and the like. The choice of technique will depend upon the target plant to be transformed. For example, dicotyledonous plants and certain monocots and gymnosperms may be transformed by Agrobacterium Ti plasmid technology, as described, for example by Bevan (Nucl. Acid Res. 12:8711-8721, 1984). Targets for the introduction of the DNA constructs of the present invention include tissues, such as leaf tissue, disseminated cells, protoplasts, seeds, embryos, meristematic regions; cotyledons, hypocotyls, and the like. One preferred method for transforming eucalyptus and pine is a biolistic method using pollen (see, for example, Aronen 1996, Finnish Forest Res. Papers vol. 595, 53pp) or easily regenerable embryonic tissues. Other transformation techniques which may be usefully employed in the inventive methods include those taught by Ellis et al. (Plant WO 98/11205 PCT/NZ97/00112 Cell Reports, 8:16-20, 1989), Wilson et al. (Plant Cell Reports 7:704-707, 1989) and Tautorus et al. (Theor. Appl. Genet. 78:531-536, 1989).

Once the cells are transformed, cells having the inventive DNA construct incorporated in their genome may be selected by means of a marker, such as the kanamycin resistance marker discussed above. Transgenic cells may then be cultured in an appropriate medium to regenerate whole plants, using techniques well known in the art. In the case of protoplasts, the cell wall is allowed to reform under appropriate osmotic conditions. In the case of seeds or embryos, an appropriate germination or callus initiation medium is employed. For explants, an appropriate regeneration medium is used. Regeneration of plants is well established for.many species. For a review of regeneration of forest trees see Dunstan et al., Somatic embryogenesis in woody plants. In: Thorpe, T.A. ed., 1995: in vitro embryogenesis of plants. Vol. 20 in Current Plant Science and Biotechnology in Agriculture, Chapter 12, pp. 471-540.

Specific protocols for the regeneration of spruce are discussed by Roberts et al., (Somatic Embryogenesis of Spruce. In: Synseed. Applications of synthetic seed to crop improvement. Redenbaugh, ed. CRC Press, Chapter 23, pp. 427-449, 1993). The resulting transformed plants may be reproduced sexually or asexually, using methods well known in the art, to give successive generations of transgenic plants.

As discussed above, the production of RNA in target plant cells can be controlled by choice of the promoter sequence, or by selecting the number of functional copies or the site of integration of the DNA sequences incorporated into the genome of the target plant host. A target plant may be transformed with more than one DNA construct of the present invention, thereby modulating the lignin biosynthetic pathway for the activity of more than one enzyme, affecting enzyme activity in more than one tissue or affecting enzyme activity at more than one expression time. Similarly, a DNA construct may be assembled containing more than one open reading frame coding for an enzyme encoded by a DNA sequence of the present invention or more than one noncoding region of a gene coding for such an enzyme. The DNA sequences of the present inventive may also be employed in combination with other known sequences encoding enzymes involved in the lignin biosynthetic pathway. In this manner, it may be possible to add a lignin biosynthetic pathway to a non-woody plant to produce a new woody plant.

WO 98/11205 PCT/NZ97/00112 The isolated DNA sequences of the present invention may also be employed as probes to isolate DNA sequences encoding enzymes involved in the lignin synthetic pathway from other plant species, using techniques well known to those of skill in the art.

The following examples are offered by way of illustration and not by way of limitation.

Example 1 Isolation and Characterization of cDNA Clones from Eucalyptus grandis Two Eucalyptus grandis cDNA expression libraries (one from a mixture of various tissues from a single tree and one from leaves of a single tree) were constructed and screened as follows.

mRNA was extracted from the plant tissue using the protocol of Chang et al.

(Plant Molecular Biology Reporter 11:113-116 (1993)) with minor modifications.

Specifically, samples were dissolved in CPC-RNAXB (100 mM Tris-CI, pH 8,0; mM EDTA; 2.0 M NaCl; 2%CTAB; 2% PVP and 0.05% Spermidine*3 HCl)and extracted with Chloroform:isoamyl alcohol, 24:1. mRNA was precipitated with ethanol and the total RNA preparate was purified using a Poly(A) Quik mRNA Isolation Kit (Stratagene, La Jolla, CA). A cDNA expression library was constructed from the purified mRNA by reverse transcriptase synthesis followed by insertion of the resulting cDNA clones in Lambda ZAP using a ZAP Express cDNA Synthesis Kit (Stratagene), according to the manufacturer's protocol. The resulting cDNAs were packaged using a Gigapack II Packaging Extract (Stratagene) employing 1 pl of sample DNA from the I ligation mix. Mass excision of the library was done using XLI -Blue MRF' cells and XLOLR cells (Stratagene) with ExAssist helper phage (Stratagene). The excised phagemids were diluted with NZY broth (Gibco BRL, Gaithersburg, MD) and plated out onto LB-kanamycin agar plates containing X-gal and isopropylthio-beta-galactoside

(IPTG).

Of the colonies plated and picked for DNA miniprep, 99% contained an insert suitable for sequencing. Positive colonies were cultured in NZY broth with kanamycin and cDNA was purified by means of alkaline lysis and polyethylene glycol (PEG) precipitation. Agarose gel at 1% was used to screen sequencing templates for WO 98/11205 PCT/NZ97/00112 chromosomal contamination. Dye primer sequences were prepared using a Turbo Catalyst 800 machine (Perkin Elmer/Applied Biosystems, Foster City, CA) according to the manufacturer's protocol.

DNA sequence for positive clones was obtained using an Applied Biosystems Prism 377 sequencer. cDNA clones were sequenced first from both the 5' end and, in some cases, also from the 3' end. For some clones, internal sequence was obtained using subcloned fragments. Subcloning was performed using standard procedures of restriction mapping and subcloning to pBluescript II SK+ vector.

The determined cDNA sequence was compared to known sequences in the EMBL database (release 46, March 1996) using the FASTA algorithm of February 1996 (version 2.0u4) (available on the Internet at the ftp site ftp://ftp.virginia.edu/pub/fastal). Multiple alignments of redundant sequences were used to build up reliable consensus sequences. Based on similarity to known sequences from other plant species, the isolated DNA sequence (SEQ ID NO: 1) was identified as encoding a CAD enzyme.

In further studies, using the procedure described above, cDNA sequences encoding the following Eucalyptus grandis enzymes were isolated: PAL (SEQ ID NO: 16); C4H (SEQ ID NO: 17); C3H (SEQ ID NO: 18); F5H (SEQ ID NO: 19-21); OMT (SEQ ID NO: 22-25); CCR (SEQ ID NO: 26-29); CAD (SEQ ID NO: 30); CGT (SEQ ID NO: 31-33); CBG (SEQ ID NO: 34); PNL (SEQ ID NO: 35, 36); LAC (SEQ ID NO: 37-41); and POX (SEQ ID NO: 42-44).

Example 2 Isolation and Characterization of cDNA Clones from Pinus radiata a) Isolation of cDNA clones by high through-put screening A Pinus radiata cDNA expression library was constructed from xylem and screened as described above in Example 1. DNA sequence for positive clones was obtained using forward and reverse primers on an Applied Biosystems Prism 377 sequencer and the determined sequences were compared to known sequences in the database as described above.

WO 98/11205 PCT/NZ97/00112 Based on similarity to known sequences from other plant species, the isolated DNA sequences were identified as encoding the enzymes C4H (SEQ ID NO: 2 and 3), C3H (SEQ ID NO: PNL (SEQ ID NO: OMT (SEQ ID NO: CAD (SEQ ID NO: CCR (SEQ ID NO: PAL (SEQ ID NO: 9-11) and 4CL (SEQ ID NO: 12).

In further studies, using the procedure described above, additional cDNA clones encoding the following Pinus radiata enzymes were isolated: PAL (SEQ ID NO: 47); C4H (SEQ ID NO: 48, 49); C3H (SEQ ID NO: 50-52); OMT (SEQ ID NO: 53- 4CL (SEQ ID NO: 56, 57); CCR (SEQ ID NO: 58-70); CAD (SEQ ID NO: 71); CGT (SEQ ID NO: 72); CBG (SEQ ID NO: 73-80); PNL (SEQ ID NO: 81); LAC (SEQ ID NO: 82-84); and POX (SEQ ID NO: 85-88).

b) Isolation of cDNA clones by PCR Two PCR probes, hereinafter referred to as LNB010 and LNB011 (SEQ ID NO: 14 and 15, respectively) were designed based on conserved domains in the following peroxidase sequences previously identified in other species: vanpox, hvupox6, taepox, hvupoxl, osapox, ntopox2, ntopoxl, lespox, pokpox, luspox, athpox, hrpox, spopox, and tvepox (Genbank accession nos. D11337, M83671, X56011, X58396, X66125, J02979, D11396, X71593, D11102, L07554, M58381, X57564, Z22920, and Z31011, respectively).

RNA was isolated from pine xylem and first strand cDNA was synthesized as described above. This cDNA was subjected to PCR using 4 [M LNB010, 4 CiM LNB011, 1 x Kogen's buffer, 0.1 mg/ml BSA, 200 mM dNTP, 2 mM Mg 2 and 0.1 U/l of Taq polymerase (Gibco BRL). Conditions were 2 cycles of 2 min at 94 1 min at 55 "C and 1 min at 72 25 cycles of 1 min at 94 1 min at 55 and 1 min at 72 and 18 cycles of 1 min at 94 OC, 1 min at 55 and 3 min at 72 "C in a Stratagene Robocycler. The gene was re-amplified in the same manner. A band of about 200 bp was purified from a TAE agarose gel using a Schleicher Schuell Elu- Quik DNA purification kit and clones into a T-tailed pBluescript vector (Marchuk D. et al., Nucleic Acids Res. 19:1154, 1991). Based on similarity to known sequences, the isolated gene (SEQ ID NO: 13) was identified as encoding pine peroxidase (POX).

WO 98/11205 PCT/NZ97/00112 Example 3 Use of an O-methvltransferase (OMT) Gene to Modify Lignin Biosynthesis a) Transformation of tobacco plants with a Pinus radiata OMT gene Sense and anti-sense constructs containing a sequence including the coding region of OMT (SEQ ID NO: 53) from Pinus radiata were inserted into Agrobacterium tumefaciens LBA4301 (provided as a gift by Dr. C. Kado, University of California, Davis, CA) by direct transformation using published methods (see, An G, Ebert PR, Mitra A, Ha SB: Binary Vectors. In: Gelvin SB, Schilperoort RA (eds) Plant Molecular Biology Manual, Kluwer Academic Publishers, Dordrecht (1988)). The presence and integrity of the transgenic constructs were verified by restriction digestion and DNA sequencing.

Tobacco (Nicotiana tabacum cv. Samsun) leaf sections were transformed using the method of Horsch et al. (Science, 227:1229-1231, 1985). Five independent transformed plant lines were established for the sense construct and eight independent transformed plant lines were established for the anti-sense construct for OMT.

Transformed plants containing the appropriate lignin gene construct were verified using Southern blot experiments. A in the column labeled "Southern" in Table 1 below indicates that the transformed plant lines were confirmed as independent transformed lines.

b) Expression of Pinus OMT in transformed plants Total RNA was isolated from each independent transformed plant line created with the OMT sense and anti-sense constructs. The RNA samples were analysed in Northern blot experiments to determine the level of expression of the transgene in each transformed line. The data shown in the column labeled "Northern" in Table 1 shows that the transformed plant lines containing the sense and anti-sense constructs for OMT all exhibited high levels of expression, relative to the background on the Northern blots.

OMT expression in sense plant line number 2 was not measured because the RNA sample showed signs of degradation. There was no detectable hybridisation to RNA samples from empty vector-transformed control plants.

WO 98/11205 PCT/NZ97/00112 c) Modulation of OMT enzyme activity in transformed plants The total activity of OMT enzyme, encoded by the Pinus OMT gene and by the endogenous tobacco OMT gene, in transformed tobacco plants was analysed for each transformed plant line created with the OMT sense and anti-sense constructs. Crude protein extracts were prepared from each transformed plant and assayed using the method of Zhang et al. (Plant Phvsiol., 113:65-74, 1997). The data contained in the column labeled "Enzyme" in Table 1 shows that the transformed plant lines containing the OMT sense construct generally had elevated OMT enzyme activity, with a maximum of 199%, whereas the transformed plant lines containing the OMT anti-sense construct generally had reduced OMT enzyme activity, with a minimum of relative to empty vector-transformed control plants. OMT enzyme activity was not estimated in sense plant line number 3.

d) Effects of Pinus OMT on lignin concentration in transformed plants The concentration of lignin in the transformed tobacco plants was determined using the well-established procedure of thioglycolic acid extraction (see, Freudenberg et al. in "Constitution and Biosynthesis of Lignin", Springer-Verlag, Berlin, 1968).

Briefly, whole tobacco plants, of an average age of 38 days, were frozen in liquid nitrogen and ground to a fine powder in a mortar and pestle. 100 mg of frozen powder from one empty vector-transformed control plant line, the five independent transformed plant lines containing the sense construct for OMT and the eight independent transformed plant lines containing the anti-sense construct for OMT were extracted individually with methanol, followed by 10% thioglycolic.acid and finally dissolved in 1 M NaOH. The final extracts were assayed for absorbance at 280 nm. The data shown in the column labelled "TGA" in Table 1 shows that the transformed plant lines containing the sense and the anti-sense OMT gene constructs all exhibited significantly decreased levels of lignin, relative to the empty vector-transformed control plant lines.

WO 98/11205 PCT/NZ97/00112 Table 1 olant line transwene nrientatinn S i 4 t'rn Nnrthprn Fvrn-Nm TrA A-Ar htr Irrl L -Jfllr I plant lne tra went-.nrif-nttion Suthern Nnrth -n Pn m- Try 1 1 2 4 1 2 3 4 6 7 8 control

OMT

OMT

OMT

OMT

OMT

OMT

OMT

OMT

OMT

OMT

OMT

OMT

OMT

na sense sense sense.

sense sense anti-sense anti-sense anti-sense anti-sense anti-sense anti-sense anti-sense anti-sense blank 2.9E+6 na 4.1E+6 2.3E+6 3.6E+5 1.6E+4 5.7E+3 8.0E+3 1.4E+4 2.5E+4 2.5E+4 2.5E+4 1.1E+4 These data clearly indicate that lignin concentration, as measured by the TGA assay, can be directly manipulated by either sense or anti-sense expression of a lignin biosynthetic gene such as OMT.

Example 4 Use of a 4-Coumarate:CoA ligase (4CL) Gene to Modify Lignin Biosynthesis a) Transformation of tobacco plants with a Pinus radiata 4CL gene Sense and anti-sense constructs containing a sequence including the coding region of 4CL (SEQ ID NO: 56) from Pinus radiata were inserted into Agrobacterium tumefaciens LBA4301 by direct transformation as described above. The presence and integrity of the transgenic constructs were verified by restriction digestion and DNA sequencing.

Tobacco (Nicotiana tabacum cv. Samsun) leaf sections were transformed as described above. Five independent transformed plant lines were established for the sense construct and eight independent transformed plant lines were established for the anti-sense construct for 4CL. Transformed plants containing the appropriate lignin gene construct were verified using Southern blot experiments. A in the column WO 98/11205 PCT/NZ97/00112 labeled "Southern" in Table 2 indicates that the transformed plant lines listed were confirmed as independent transformed lines.

b) Expression ofPinus 4CL in transformed plants Total RNA was isolated from each independent transformed plant line created with the 4CL sense and anti-sense constructs. The RNA samples were analysed in Northern blot experiments to determine the level of expression of the transgene in each transformed line. The data shown in the column labelled "Northern" in Table 2 below shows that the transformed plant lines containing the sense and anti-sense constructs for lo 4CL all exhibit high levels of expression, relative to the background on the Northern blots. 4CL expression in anti-sense plant line number 1 was not measured because the RNA was not available at the time of the experiment. There was no detectable hybridisation to RNA samples from empty vector-transformed control plants.

c) Modulation of 4CL enzyme activity in transformed plants The total activity of 4CL enzyme, encoded by the Pinus 4CL gene and by the endogenous tobacco 4CL gene, in transformed tobacco plants was analysed for each transformed plant line created with the 4CL sense and anti-sense constructs. Crude protein extracts were prepared from each transformed plant and assayed using the method of Zhang et al. (Plant Physiol., 113:65-74, 1997). The data contained in the column labeled "Enzyme" in Table 2 shows that the transformed plant lines containing the 4CL sense construct had elevated 4CL enzyme activity, with a maximum of 258%, and the transformed plant lines containing the 4CL anti-sense construct had reduced 4CL enzyme activity, with a minimum of 59%, relative to empty vector-transformed control plants.

d) Effects of Pinus 4CL on lignin concentration in transformed plants The concentration of lignin in samples of transformed plant material was determined as described in Example 3. The data shown in the column labelled "TGA" in Table 2 shows that the transformed plant lines containing the sense and the antisense 4CL gene constructs all exhibited significantly decreased levels of lignin, relative to the empty vector-transformed control plant lines. These data clearly indicate that WO 98/11205 PCT/NZ97/00112 lignin concentration, as measured by the TGA assay, can be directly manipulated by either sense or anti-sense expression of a lignin biosynthetic gene such as 4CL.

Table 2 plant line transeene orientation Southern Northern Enzyme TGA Northern En7vme T A 1 2 1 2 3 4 5 1 2 3 4 5 6 7 8 control control 4CL 4CL 4CL 4CL 4CL 4CL 4CL 4CL 4CL 4CL 4CL 4CL 4CL na na sense sense sense sense sense anti-sense anti-sense anti-sense anti-sense anti-sense anti-sense anti-sense anti-sense blank blank 2.3E+4 4.5E+4 3.1E+4 1.7E+4 1.6E+4 na 1.0E+4 9.6E+3 1.2E+4 4.7E+3 3.9E+3 1.8E+3 1.7E+4 100 100 169 258 174 164 184 59 81 101 116 125 106 Example Transformation of Tobacco using the Inventive Lignin Biosynthetic Genes Sense and anti-sense constructs containing sequences including the coding regions of C3H (SEQ ID NO: 18), F5H (SEQ ID NO: 19), CCR (SEQ ID NO: 25) and CGT (SEQ ID NO: 31) from Eucalyptus grandis, and PAL (SEQ ID NO: 45 and 47), C4H (SEQ ID NO: 48 and 49), PNL (SEQ ID NO: 81) and LAC (SEQ ID NO: 83) from Pinus radiata were inserted into Agrobacterium tumefaciens LBA4301 by direct transformation as described above. The presence and integrity of the transgenic constructs were verified by restriction digestion and DNA sequencing.

Tobacco (Nicotiana tabacum cv. Samsun) leaf sections were transformed as described in Example 3. Up to twelve independent transformed plant lines were established for each sense construct and each anti-sense construct listed in the preceding paragraph. Transformed plants containing the appropriate lignin gene WO 98/11205 PCT/NZ97/00112 construct were verified using Southern blot experiments. All of the transformed plant lines analysed were confirmed as independent transformed lines.

Example 6 Manipulation of Lignin Content in Transformed Plants a) Determination of transgene expression by Northern blot experiments Total RNA was isolated from each independent transformed plant line described in Example 5. The RNA samples were analysed in Northern blot experiments to determine the level of expression of the transgene in each transformed line. The column labelled "Northern" in Table 3 shows the level of transgene expression for all plant lines assayed, relative to the background on the Northern blots. There was no detectable hybridisation to RNA samples from empty vector-transformed control plants.

b) Determination of lignin concentration in transformed plants The concentration of lignin in empty vector-transformed control plant lines and in up to twelve independent transformed lines for each sense construct and each anti-sense construct described in Example 5 was determined as described in Example 3. The column labelled "TGA" in Table 3 shows the thioglycolic acid extractable lignins for all plant lines assayed, expressed as the average percentage of TGA extractable lignins in transformed plants versus control plants. The range of variation is shown in parentheses.

WO 98/11205 PCT/NZ97/00112 Table 3 transgene orientation no. of lines Northern TGA control na 3 blank 100 (92-104) C3H sense 5 3.7E+4 74 (67-85) sense 10 5.8E+4 70 (63-79) anti-sense 9 5.8E+4 73 (35-93) CCR sense 1 na 74 CCR anti-sense 2 na 74 (62-86) PAL sense 5 1.9E+5 77 (71-86) PAL anti-sense 4 1.5E+4 62 (37-77) C4H anti-sense 10 5.8E+4 86 (52-113) PNL anti-sense 6 1.2E+4 88 (70-114) LAC sense 5 1.7E+5 na LAC anti-sense 12 1.7E+5 88 (73-114) Transformed plant lines containing the sense and the anti-sense lignin biosynthetic gene constructs all exhibited significantly decreased levels of lignin, relative to the empty vector-transformed control plant lines. The most dramatic effects on lignin concentration were seen in the F5H anti-sense plants with as little as 35% of the amount of lignin in control plants, and in the PAL anti-sense plants with as little as 37% of the amount of lignin in control plants. These data clearly indicate that lignin concentration, as measured by the TGA assay, can be directly manipulated by conventional anti-sense methodology and also by sense over-expression using the inventive lignin biosynthetic genes.

Example 7 Modulation of Lignin Enzyme Activity in Transformed Plants The activities and substrate specificities of selected lignin biosynthetic enzymes were assayed in crude extracts from transformed tobacco plants containing sense and anti-sense constructs for PAL (SEQ ID NO: 45), PNL (SEQ ID NO: 81) and LAC (SEQ ID NO: 83) from Pinus radiata, and CGT (SEQ ID NO: 31) from Eucalyptus grandis.

Enzyme assays were performed using published methods for PAL (Southerton, S.G. and Deverall, Plant Path. 39:223-230, 1990), CGT (Vellekoop, P. et al., FEBS, 330:36-40, 1993), PNL (Espin, C.J. et al., Phvtochemistrv, 44:17-22, 1997) and WO 98/11205 PCT/NZ97/00112 LAC (Bao, W. et al., Science, 260:672-674, 1993). The data shown in the column labelled "Enzyme" in Table 4 shows the average enzyme activity from replicate measures for all plant lines assayed, expressed as a percent of enzyme activity in empty vector-transformed control plants. The range of variation is shown in parentheses.

Table 4 transgene orientation no. of lines Enzyme control na 3 100 PAL sense 5 87 (60-124) PAL anti-sense 3 53 (38-80) CGT anti-sense 1 89 PNL anti-sense 6 144 (41-279) LAC sense 5 78 (16-240) LAC anti-sense 11 64 (14-106) All of the transformed plant lines, except the PNL anti-sense transformed plant lines, showed average lignin enzyme activities which were significantly lower than the activities observed in empty vector-transformed control plants. The most dramatic effects on lignin enzyme activities were seen in the PAL anti-sense transformed plant lines in which all of the lines showed reduced PAL activity and in the LAC anti-sense transformed plant lines which showed as little as 14% of the LAC activity in empty vector-transformed control plant lines.

Example 8 Functional Identification of Lignin Biosynthetic Genes Sense constructs containing sequences including the coding regions for PAL (SEQ ID NO: 47), OMT (SEQ ID NO: 53), 4CL (SEQ ID NO: 56 and 57) and POX (SEQ ID NO: 86) from Pinus radiata, and OMT (SEQ ID NO: 23 and 24), CCR (SEQ ID NO: 26-28), CGT (SEQ ID NO: 31 and 33) and POX (SEQ ID NO: 42 and 44) from Eucalyptus grandis were inserted into the commercially available protein expression vector, pProEX-1 (Gibco BRL). The resultant constructs were transformed into E. coli XL1-Blue (Stratagene), which were then induced to produce recombinant protein by the addition of IPTG. Purified proteins were produced for the Pinus OMT and 4CL constructs and the Eucalyptus OMT and POX constructs using Ni column 24 WO 98/11205 PCT/NZ97/00112 chromatography (Janknecht, R. et al., Proc. Natl. Acad. Sci., 88:8972-8976, 1991).

Enzyme assays for each of the purified proteins conclusively demonstrated the expected substrate specificity and enzymatic activity for the genes tested.

The data for two representative enzyme assay experiments, demonstrating the verification of the enzymatic activity of a Pinus radiata 4CL gene (SEQ ID NO: 56) and a Pinus radiata OMT gene (SEQ ID NO: 53), are shown in Table 5. For the 4CL enzyme, one unit equals the quantity of protein required to convert the substrate into product at the rate of 0.1 absorbance units per minute. For the OMT enzyme, one unit equals the quantity of protein required to convert 1 pmole of substrate to product per minute.

Table purification total ml total mg total units yield fold transgene step extract protein activity activity purification 4CL crude 10ml 51 mg 4200 100 1 Ni column 4 ml 0.84 mg 3680 88 53 OMT crude 10 ml 74 mg 4600 100 1 Ni column 4 ml 1.2 mg 4487 98 The data shown in Table 5 indicate that both the purified 4CL enzyme and the purified OMT enzyme show high activity in enzyme assays, confirming the identification of the 4CL and OMT genes described in this application. Crude protein preparations from E. coli transformed with empty vector show no activity in either the 4CL or the OMT enzyme assay.

Although the present invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, changes and modifications can be carried out without departing from the scope of the invention which is intended to be limited only by the scope of the appended claims.

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(ii) TITLE OF THE INVENTION: MATERIALS AND METHODS FOR THE MODIFICATION OF PLANT LIGNIN CONTENT (iii) NUMBER OF SEQUENCES: 88 (iv) CORRESPONDENCE

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INFORMATION FOR SEQ ID NO:1: SEQUENCE CHARACTERISTICS: LENGTH: 535 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1: CTTCGCGCTA CCGCATACTC CACCACCGCG TGCAGAAGAT GAGCTCGGAG GGTGGGAAGG AGGATTGCCT CGGTTGGGCT GCCCGGGACC CTTCTGGGTT CCTCTCCCCN TACAAATTCA 120 CCCGCAGGCC GTGGGAAGCG AAGACGTCTC GATTAAGATC ACGCACTGTG GAGTGTGCTA 180 CGCAGATGTG GCTTGGACTA GGAATGTGCA GGGACACTCC AAGTATCCTC TGGTGCCGGG 240 WO 98/11205 WO 9811205PCT/NZ97/001 12 GCACGAGATA GTTGGAATTG TGAAACAGGT TGGCTCCAGT GTCCAACGCT TCAAAGTTGG 300 CGATCATGTG GGGGTGGGAA CTTATGTCAA TTCATGCAGA GAGTGCGAGT ATTGCAATGA 360 CAGGCTAGAA GTCCAATGTG AAAAGTCGGT TATGACTTTT GATGGAATTG ATGCAGATGG 420 TACAGTGACA AAGGGAGGAT ATTCTAGTCA CATTGTCGTC CATGAAAGGT ATTGCGTCAG 480 GATTCCAGAA AACTACCCGA TGGATCTAGC AGCGCATTGC TCTGTGCTGG ATCAC 535 INFORMATION FOR SEQ ID NO:2: SEQUENCE CHARACTERISTICS: LENGTH: 671 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

GCGCCTGCAG

TGATTTGAAT

120

CAAGATGGGC

180

GCACACCCAG

240

GGGCAAGGGG

300

GATCATGACT

360

AGACGAGATC

420

CATTGTCATC

480

GACAGGAGAT

540

GAGCGAAGTC

600

AGGCCCTTCC

660

CTTTTCAAGG

671 SEQUENCE DESCRIPTION: SEQ ID NO:2: GTCGACACTA GTGGATCCAA AGAATTCGGC

CACAGAAACC

CAGAGGAATC

GGCGTCGAGT

CAGGACATGG

GTGCCTTTCT

AGCCGCGTGG

CGTAGCGCCT

TCGAATCCGA

GATTGGCCCA

TCAGAGGTTA

TCAGCGATTT

TTGTGGTAGT

TTGGGTCTCG

TGTTCACCGT

TTACGAATAA

TCGCGGATGT

CCAGCTCATG

GGACGACCCG

GAGCTTTGAG

TCACAGAATC

TGCCAAGAAA

TTCATCTCCC

AACCCGGAAC

CTATGGAGAT

AGTTGTCCAG

GAAATCCCGC

ATGTATAATA

CTTTTCCTCA

TACAAT TAT G

TGCAATGAGA

ACGAGGTTGC

TATGGCAAAA

GATCTCGCCA

GTGGTGTTCG

CACTGGAGAA

CACTACAGAT

GCCGAGTCTT

TTATGTATAG

AGCTCAAGGC

GGGATTTCAT

TTAAAGAGAA

AGGTCGGGGA

TCTTTCTGCT

AGGAGGTCCT

ATATCTTCAC

AGATGCGCAG

TCGCGTGGGA

CCACCTCGGG

GATGATGTTC

CCTCAACGGA

TCCCAGTCTT

ACGGCTCTCT

INFORMATION FOR SEQ ID NO:3: SEQUENCE CHARACTERISTICS: LENGTH: 940 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: CTTCAGGACA AGGGAGAGAT CAATGAGGAT GTTGCAGCAA TTGAGACAAC GCTGTGGTCG 120 CACCAGGACA TTCAGAGCAA GGTGCGCGCA 180 CAGATAACGG AACCAGACAC GACAAGGTTG 240 SEQ ID NO:3: AATGTTTTGT ACATCGTTGA GAACATCAAC ATGGAATGGG GAATAGCGGA GCTGGTGAAC GAGCTGGACG CTGTTCTTGG ACCAGGCGTG CCCTACCTTC AGGCGGTTGT GAAGGAAACC WO 98/11205 WO 98/ 1205PCT/NZ97/00112 CT TCGTCT CC 300

CTCGGGGGCT

360

AACAACCCCG

420

GAGAAGCACA

480

AGGAGCTGCC

540

TTCAGAACTT

600

GCGGGCAATT

660

CTGCTTAATC

720

CTCCATCTAT

780

CTTCAAAAGT

840

AAGTTTGCAT

900

ATTTTACTGC

940

GCATGGCGAT

ACGATATTCC

CCAACTGGAA

CCGAAGCCAA

CGGGAATCAT

CCACCTTCTG

CAGCCTTCAC

CCAACTTGTC

CATGACTGTG

TTGCTAGGAT

AAATTAAATG

TAAAAAAAAA

CCCGTTGCTC

GGCAGAGAGC

GAACCCCGAG

TGGCAACGAC

TCTGGCGCTG

CCGCCGCCCG

ATTCTCAACC

AGTGACTGGT

TGTGCGTGTC

TTCAATAACA

ATATTTCAAT

GTCCCCCACA

AAGATCCTGG

GAG TT C CCCC

TTCAAATTCC

CTCTCCTCGC

GGCAGAGCAA

ATTCTCTCAT

ATATAAATGC

CACTGTCGAG

GACACCGTCA

ATACTATTTT

AAAAAAAA

TGAATCTCCA

TGAACGCCTG

CCGAGCGGTT

TGNCCTTCGG

ACTCTCCATC

AGTGGATGTC

CGTCGCCAAG

GCGCACCTGA

TCTACTAAGA

ATTATGTCAT

GACTCTCCAC

CGACGCCAAG

GTGGTTGGCC

CTTCGAGGAG

TGTGGGGAGG

GGAAGACTTG

ACTGAGAAGG

CCCATAGCTT

ACAAAAAACA

GCTCATAGCA

GTTTCAATAA

CAATTGGGGA

INFORMATION FOR SEQ ID NO:4: SEQUENCE CHARACTERISTICS: LENGTH: 949 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

NNGCTCNACC

CAGGCAATTC

120

CTCGTATGTT

180

ATGATTACGC

240

GCGGTGGCGG

300

TTCAGGCCTG

360

CTGCCATTGG

420

AGTCTATGTT

480

CAGAAGACAT

540

AGGCCATTGC

600

CAATTGATCT

660

TCTCCTTTTC

720

TCTGAAGCCC

780

AAATTGAGWA

840

TANTTTAGGG

900 SEQUENCE DESCRIPTION: SEQ ID NO:4: GACGGTGGAC GGTCCGCTAC

GATTTAGCTC

GTGTGGAATT

CAAGCGCGCA

CCGCTCTAGA

AGAGATTTCT

TGCAGGGCGC

GGGACACCTG

AGATCTCACA

TATTCCTCGA

GATAGTAAGT

CATAGTCAAC

AACTTCTAGC

ACTCATTAGG

CT GAGCG CAT

ATTAACCCTC

ACTAGTGGAT

TGAGGAAGAT

AGGATCTGCC

CTTCATCATT

GAGAATCCAG

TTGCCTGATC

TTGAATTTTG

ATGCAGCTTT

AAGCAATAAC

TCAGTAACTG

CACCCCAGGC

AACAATTTCA

ACTAAAGGGA

CCAAAGAATT

GTTGATATTA

CTGGTGCACA

TCGTATGGGC

GGCTTGTTAC

ATCTCTACAA

TTTTGATACA

CTTTCTCTGA

TGTATATTTT

ATTGTGCAAT

ANANGNGGNA

AGTGGGATCC

TTTACACTTT

CACAGGAAAC

ACAAAAGCTC.

CGGCACGAGA

AGGCCCATGA

ATTGGGTATT

ACCTCCTGAG

TTTCATGGCC

GCGACAGCCA

AAACGAAATA

AGCGCATGCA

AGAACAAATA

TTGCAAGNAA

ATGNTAGNGG

CCCGGGCTGA

ATGCTTCCGG

AGCTATGACC

GAGCTCCACC

CCCAGTGACC

TTACAGGCTA

AATTTAGTTC

GGAATGAAGG

AAGCCTGTGC

CTCAATTGAT

ACGTGCAGTT

GCTTTCTTTC

CCTATTCCTC

TAGTAAAGTT

GCATTNAGAA

TTTCTCTGTA GGGGNNGNTA NATTTTAATA GTCCTANGTA WO 98/11205 WO 9811205PCT/NZ97/O01 12 ANCCCTAATA GNTGTTGGNG GNNGNTAGGN TTTTTNACCA AAAAAAAAA 949 INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 959 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID GAATTCGGCA CGAGAAAGCC CTAGAATTTT

CTTTAACTGC

120

CTGGTCAGGT

180

CTTGCACTCA

240

TGGAAGCTTG

300

TCAAGCAAGT

360

TGACCCTCTC

420

AACTCCATGT

480

ATGGGATGGT

540

ATATAGATAC

600

AGCAGATTTC

660

ACATAGATGA

720

TCATCTTCAA

780

TAGTACTGTG

840

AAAATCTCAA

900

TGACATTTGA

959

AATAACTGTG

TCCAGCATTT

GCCCTACATT

TGTCAACACG

T TTCT CAT CT

TCTTCAAGAC

TCTGCAGATG

GAGCTTCAAT

TGCAATTCGG

GTGTGATCAT

TTTTGATACT

GACTCGCTTA

GCTGAGTCCA

ATTTCTCGAT

GCACCTCGAG

GAAGCGTACA

CCAAAATACA

GATTTAGCAA

AACACAGAGA

CTTTATAAAC

ATAGCAAGTA

ATTCAAGATG

GAGGATCCTG

AGAATCATGG

TCCTACCTGA

GTTCCCCAGA

TATTCATTAC

GAAAGGATCT

GTCTAGTCTT

TGAACTACAA

TTCAGCATGC

AAAAGTTTGT

CACCTGCTGT

ACAACTACAG

AGTTCAAGAA

GGAATATTCA

CGGTACAGTT

GTGAGATTTT

AACAGTACAA

CACTATCAAA

GTAAGGTGGG

AGTTCANAAA

TTTCTATGTG

CTCGGTATTA

GATTTTGATT

AGTTGCATGT

TATCACAGCC

CCTAGTTTCT

TGTCCAAAGA

TAGTGGGAAA

TGATAGTAAT

GAGATTGACA

GGAGACTGCT

TGCAACCATA

AACATGTCAG

GAAGCTCACC

GAGAGAGCGT

TATGTAACAA

AATTGATAGT

TCACTTGACA

ATGAATGCGA

TAAAAAAAAA

CCAGCGACAA

CTCATTCAGA

AATTTGAAAT

ATTTCTGTAT

TTGGGGTTAG

CAGACATATC

AAGCAGGCTG

AATCAGAAAG

ATGACTGAAT

ACAGTAGATG

TCAAGATTTG

ATGATGTAAA

CTGTTAACAA

TGCCATCAAA

CTTTTAGTTG

INFORMATION FOR SEQ ID NO:6: SEQUENCE CHARACTERISTICS: LENGTH: 1026 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: GAATTCGGCA CGAGCTTTGA GGCAACCTAC CAAACAGGTT TAAGGAAATG GCAGGCACA 120 CAACCCAAGC AGAGGAGCCG GTTAAGGTTG 180 TTTTGCAGAG CGATGCCCTC TATCAGTATA 240 SEQ ID NO:6: ATTCATTGAA TCCCAGGATT TCTTCTTGTC GTGTTGCTGC AGCAGAGGTG AAGGCTCAGA TCCGCCATCA AGAAGTGGGA CACAAAAGTC TATTGGAAAC GAGCGTGTAC CCTCGTGAGC WO 98/11205

CCGAGCCAAT

300

CTTCTGCCGA

360

CCATGGAGAT

420

ATGATGGAAA

480

TTATTGAGAA

540

TTCTGGACGA

600

ATGCGGACAA

660

GAGGTCTGAT

720

CTCCCCTGAG

780

TTGCTGTCGA

840

GCAGGCGTGT

900

CTCTGATTAT

960

TATTGATAAT

1020 1026

GAAGGAGCTC

TGAGGGTCAA

TGGGGTGTAC

GATTCTAGCC

AGCAGGAGTT

ACTGCTTAAG

AGACAACTAT

TGCATATGAC

GAAATATGTG

TCCCCGCATT

CTATTGAAAA

AAGGAGAACG

AAAGTAGTAC

CGCGAAGTGA

TTTCTGGGCC

ACTGGTTACT

ATGGACATCA

GCCCACAAGA

AATGAGGACA

CTAAACTACC

AACACCCTGT

AGATATTACA

GAGATCAGCC

CAATCCTTGT'

CTATAATATA

AGCATATGCA

CTGCCAAGCA

TCCTGCTGAA

CGCTTCTCAG

ACAGAGAGAA

TTGACTTCAG

TGCATGGATC

ACAAGCGTCT

GGAACGGATC

GAGATTTCGT

AAATCCCAGT'

TTCTGCTCGT

TGGGGTTGAA

AAGTTTGTAT

TCCCTGGAAC

GCTCATTAAC

CACAGCCCTT

CTATGATATC

AGAGGGCCCT

GTT CGATTTT

GATCGATCTG

TGTGGTGGCT

GATGGAGCTA

CGGTGACGGC

CTATTGCAAG

GCCATTTGTT

CAAAAAAAAA

PCT/NZ97/00112

CTCATGACTA

GCCAAGAACA

GCATTGCCCG

GGATTGCCTA

GCTCTGCCAG

GTGTTCGTGG

GTGAAGGTTG

CCACCCGATG

AACAAGGCCC

GTCACCCTTT

CATAAAGGCT

TTGTTTAGTG

AAAMAA

INFORMATION FOR SEQ ID NO:7: SEQUENCE CHARACTERISTICS: LENGTH: 1454 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7: GAATTCGGCA CGAGGCCAAC TGCAAGCAAT

TGAAGTGGTT

120

GCTCGGGACT

180

GAGGATGTAA

240

CGTAATGAAA

300

GTAACAGAGA

360

TGCATTGTTG

420

AGCAAGAGGA

480

TTTGCAAGCA

540

CTGGAACAAG

600

TTCGCCATGA

660

ATGGGTGTCA

720

AAAAAGAAAG

780

CTGAAGTGAT

CCAGTGGCCA

TTGTAAAGGT

TGGACATGTC

TTGGCAGCGA

GGTCCTGTCG

TTTGGACCTA

GTATGGTGGT

CGGCCCCTCT

CAGAGCCCGG

AGATTGCCAA

AAGAAGCCAT

GGGAAGCTTG

CTTGTCCCCT

CATTTACTGC

TCATTACCCA

GGTGAAGAAA

CAGTTGCGGT

CAATGATGTG

TGATCAGATG

GTTATGTGCA

GAAGAAATGT

AGCCTTTGGA

GGAAGTCCTC

ACAGTACAAG

GAATCTGAAA.

TACACTTACA

GGAATCTGCC

ATGGTCCCTG

TTCAAAGTGG

AATTGCAATC

AACCATGACG

TWTGTGGTTC

GGGTTACAG

GGGATTTTGG

CTCCACGTGA

GGCGCCGATG

AGCCAGACGA

AAACTGTTAC

ATCTCAGAAA

ACTCTGATTT

GGCATGAAGT

GAGAGCATGT

AGAGCATGGA

GCACACCTAC

GAATCCCGGA

TTTTCAGCCC

GTTTAGGAGG

CGGTTATCAG

CTTATCTTGT

TCGAATCCTG

AGGATATGCA

GAAAGGACCT

AGTTCAAATG

GGTGGGGATT

AGGGGTTGGT

ACAATACTGC

TCAGGGCGGA

GAATCTTCCT

AATGAAGCAT

CGTGGGGCAC

TTCGTCTGAT

TAGCAAGGAT

WO 98/11205 WO 9811205PCT/NZ97/001 12 ACT GAAAAGA 840

GCTCATCCTC

900

GGCGTTGTTC

960

ATAGCTGGAA

1020

GAGAAGAAGG

1080

GAAAGGTTGG

1140

TTGGATAATT

1200

CTGGACTAGT

1260

TTTTTGTTAC

1320

GTATATGTAA

1380

TAATATATGT

1454 AAAAAA 1454

TGATGGAAGC

TGGAACCATA

CAGAGTCGTT

GTTTCATTGG

TATCATCGAT

AGAAGAACGA

AGTCTGCAAT

AGCTTAACAT

TTTAGTTTAG

AGATCAATTT

ATTCGTATTT

AGCAGAGAGC

TCTTGCCCTT

GCACTTCGTG

CAGCATGGAG

GATTGAGGTT

TGTCCGTTAC

CAATCAATCA

GAAAGGGAAA

CTTTTGTGAG

CTCGTGACAG'

CTAGATTACA

CTGAAGACAA

ACTCCTCTCT

GAAACACAGG

GTGGGCCTGG

AGATTTGTGG

GATCAATGCC

TTAAATTTTT

GTTGAAACAA

TAAATAATAA

TAATGGACAC

ATGGAAAGCT

TAATACTTGG

AAACTCTAGA

ACTACATCAA

TGGATGTTGC

TGCATGCAAG

ATTTAGGAAC

TTCAGATGTT

TCCAATGTCT

CATTCCAGTT

AGTGATGCTG

GAGAAGGAGC

TTTCTGTGCA

CACGGCCATG

TAGAAGCAAG

ATGAATAGAT

TCGATACTGG

TTTT TAACTT

TCTGCCAAAT

TTATATGAAA.AAAAAA AAAA 1440 AAAAAA AAA INFORMATION FOR SEQ ID NO:8: SEQUENCE CHARACTERISTICS: LENGTH: 740 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8: GAATTCGGCA CGAGACCATT TCCAGCTAAT ATTGGCATAG CAATTGGTCA TTCTATCTTT

GTCAAAGGAG

120

TACCCAGATG

180

CGAGATTCTC

240

GCATTTGCCA

300

TTTAGTATAG

360

AACAAATACT

420

TCTTTGGAAA

480

CCAAAATCAT

540

CAGTGAATAA

600

GTGTTAGTGA

660

ATCAAACAAA

TGAAATATAC

TTCCACATGC

AATTGTGGGT

TATGACGAGC

CACCTGTGGT

CCGCTTAGTG

GGCTGATGTG

TTTTGTTAGA

ACGGAATGAT

TTTTGAAATT

CACTGTCGAT

TTCAGAGATA

TATAATCCTT

TAGGCACTGC

TTGTTTTCTT

TGGAATGCTA

AACTGGTTGT

GTGTTTAGAT

GTCAAATCTT

GGACCTAATG

GAGTACCTCA

CATAACAGTT

CGTAGGTGTT

AGATCCTTCA

TCTTTCTGGA

AGTACTAGTG

TCCAGAGGGT

CCATCTTTAC

GATGGGCTGA

TAAA.AAAAAA

GTGTGGAGGC

GCAAATTTGT

TCAATCAATG

TGGCAGAACA

CACTTTTCTC

ACTTTGGTAT

TCCAGAGTTC

GTTTACAACC

AAGGCTATTG

CTGACTCTCT

AAAAAA

TAGtCAGCTA

GTGAAGTATG

TTTGTCCTAG

GAACCTCCTG

TTCCATAAGA

GGCAATAATG

TAAGGGAGTT

AACAGTTGTT

AGTAAGGTTG

TGTGATGTCA

AATCTTGATG GATTGTGTCT TTTTCAATGG 740 720 AAAAAAAA 740 INFORMATION FOR SEQ ID NO:9: SEQUENCE CHARACTERISTICS: LENGTH: 624 base pairs TYPE: nucleic acid WO 98/11205 PCT/NZ97/00112 STRANDEDNESS: single TOPOLOGY: linear (xi)

GAATTCCTGC

GCGCGCCTGC

120

TTGTTGGACG

180

CCCAAGGAAG

240

GTCTGTGTTG

300

GAGGTGATGC

360

CCAGGGCAGA

420

AAAGAAGCAG

480

GCTCTGCGAA

540

CACTCCATCG

600

GACATGGCTG

624 SEQUENCE DESCRIPTION: SEQ ID NO:9: AGCCCGGGGG ATCCACTAGT TCTAGAGCGG CCGCCACCGC

AGGTCGACAC

CCATGGAAGC

GACTGGCTCT

ACGCCAACGT

AAGGGAAACC

TCGAAGCCGC

CGCGGCTTCA

CATCGCCACA

AGCGGGAGAT

TCCACGGCGG

TAGTGGATCC

TCTCCGGAAA

CGTCAACGGC

GCTGGGCGTG

GGAGTTCGTA

GGCCGTCATG.

CGAGAAAGAC.

GTGGTTGGGG

CAATTCCGTC

CAAC

AAAGAATTCG

GCCGGGATTC

ACAGCGGTGG

CTGGCTGAGA

GATCCGTTAA

GAGTTCCTCC

CCGTTGAGCA

CCTCCGATCG

AACGACAATC

GCACGAGGCC

TGGAACCGTT

GATCCGCCGT

TTCTGTCTGC

CCCACCAGTT

TCGACGGTAG

AACCGAAACA

AAGTCATCCG

CGTTAATCGA

GGTGGAGCTC

CGACGGCCAC

TAAACTGCAG

GGCCGCGTCC

GCTCTTCTGC

GAAGCACCAC

CGACTACGTG

AGACCGCTAC

CGCTGCYACT

TGTCTCCAGG

INFORMATION FOR SEQ ID Ci) SEQUENCE CHARACTERISTICS: LENGTH: 278 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

GAATTCCTGC

CAGTACCTGG

120

GTCAATTCCC

180

CTGATGTTCG

240

,GAAAACATGC

278 SEQUENCE DESCRIPTION: SEQ ID AGCCCGGGGG ATCCACTAGT TCTAGAGCGG CCGCCACCGC GGTGGAGCTC CCAACCCCGT CACGACTCAC GTCCAGAGCG CCGAACAACA.CAACCAGGAT TCGGCTTGAT CTCCGCCAGA AAGACTGCCG AGGCCGTTGA GATTTTAAAG CTACATATCT GGTGGCCTTA TGCCAGGCGA TCGATCTCCG GCACCTGGAA GATCCGTTGT GAAGCACGTA GTCTTGCA INFORMATION FOR SEQ ID NO:11: SEQUENCE CHARACTERISTICS: LENGTH: 765 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: GAGCTCCTGC AAGTCATCGA TCATCAGCCC CCATCATACG CGCTTATGCT CCAACTCAGA 120 SEQ ID NO:l1: GTTTTCTCGT ACATCGACGA TCCCACAAAT GAAGTGCTCG TAGATGAGGC TCTCAAATCA WO 98/11205

TCTTGCCCAG

180

GCTGCTGGAA

240

AAGGCCCGTT

300

CCAATTGCAA

360

GAGTTGGGAA

420

AAGGTATTTG

480

GCTTGGGGTG

540

TTCAATGCCT

600

AGAGGTTTCT

660

CCTAAACAGC

720

GGGTTCCAAC

765

ACGGGAATGA

TATTACCCAA

TAGAGGAAGA

ACAGAATAAA

CCGATTTGCT

AGGGCATTTG

GGTGCGCTGG

CATATTGGGC

GGAGCGCCCA

TTGTTCTTCG

AAAATAGAAG

CGAATCCGAT

TTGGGTGTTT

GGTTCCGAAG

CAAGTGCAGG

AACAGGGCCC

CCAAGGGAAA

ACCATTCACT

ATGGTTTGAT

ACAACAACAA

CAATAACGAA

AAATATTTTC

CACAATTTGC

AGCAGGATCC

GCGAGGGAAC

ACATATCCCA

AAGTGGAGAA

ATTGGAAACG

CCACGTGCAT

AGCACCAAAT

GTTCTTTGAT

TCTTTCATCT

GATCCAAAAA

AGCCCGCTGA

CCATATTTCA

GATTCGATAA

TTTACAGATT

GCCCCGGCGA

TGATCCTCAA

AT CCT GCG TC

CACCCTCTGC

TTAACTGACT

TCGTTACTTT

PCTINZ97/001 12

GAGCGCTGGA

AGAGGAGTTG

TGGGGACTTC

CGTGAGATCA

AGATATAGAA

ATGTCTGGAC

TICCTGCAGCG

AACGAGCGGC

CTTAAGCATT

GTAAAAGATG

INFORMATION FOR SEQ ID NO:12: SEQUENCE CHARACTERISTICS: LENGTH: 453 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

TGATTATGCG

TGAACCTAGC

120

TCCGGAACGC

180

CAAGCCGGCG

240

GAATCCACGG

300

ATTGACGATG

360

GCTTCCAGGT

420

CGGCCGCCAC

453 SEQUENCE DESCRI PT ION: GATCCTTGGG CAGGGATACG CTTCGCAAAG AATCCTTTCC TCAAATAAAG ATCCTCGATT AAATCTGCAT CCGCGGACCC CCGCTACAAT CGATGAAGAA ACGAAGAAAT CTTCATAGTC.

GGATCCTGCT AATCGAATTC CGCGGTGGAG CTCCAGCTTT SEQ ID NO:12: GCATGACAGA AGCAGGCCCG CCGCCAAATC TGGCTCCTGC ACAGGAACTG GCGAGTCTCT GAAATAATGA AAGGATATAT GGCTGGCTCC ACACAGGCGA GACAGAGTAA. AGGAGATTAT CTGCAGCCCG GGGGTCCACT

TGT

GTGCTGGCAA

GGAACAGTCG

CCCGCACAAT

TAACGACCCG

CGTCGGGTAC

CAATATAAAG

AGTTCTAGAG

INFORMATION FOR SEQ ID NO:13: SEQUENCE CHARACTERISTICS: LENGTH: 278 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13: TCTTCGAATT CTCTTTCACG ACTGCTTCGT TAATGGCTGC GATGGCTCGA TATTGTTAGA TGATAACTCA ACGTTCACCG GAGAAAAGAC TGCAGGCCCA AATGTTAATT CTGCGAGAGG 120 WO 98/11205 PCT/NZ97/00112 ATTCGACGTA ATAGACACCA TCAAAACTCA AGTTGAGGCA GCCTGCAGTG GTGTCGTGTC 180 AGTTGCCGAC ATTCTCGCCA TTGCTGCACG CGATTCAGTC GTCCAACTGG GGGGCCCAAC 240 ATGGACGGTA CTTCTGGGAG AAAAGACGGA TCCGATCA 278 INFORMATION FOR SEQ ID NO:14: SEQUENCE CHARACTERISTICS: LENGTH: 23 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14: CTTCGAATTC WYTTYCAYGA YTG 23 INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 22 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID GATCGGATCC RTCYYKYCTY CC 22 INFORMATION FOR SEQ ID NO:16: SEQUENCE CHARACTERISTICS: LENGTH: 472 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16: AATTCGGCAC GAGACGACCT CTTGTATCGG ACCCGGATCC GCTATCGTTA ACGTACACAC

GTTCTAGTGC

120

CTGCGCCGCC

180

CACAGGGAGC

240

GCGCCTCGGC

300

GGTAGGGGTC

360

CATGGAGAGC

420

CTTCTCAAAC

472

TGAATGGAGA

GGGAGCCACC

CACCTCGACG

GGGGAGTCCC

GAGCTCTCGG

ATGAACAAGG

CGGAGGCCGA

TGGAGAGCAC

ATGCCGACCC

AGGTGAAGCG

TCACGATAGC

AGGCGGCCCG

GAACTGACAG

AGCAAGGCGG

CACCGGCACC

ACTGAACTGG

GATGGTCGAG

CCAGGTGGCG

TCCCAGGGTC

CTACGGGGTC

TCCTTTTCAG

GGCAACGGCC

GGGGCGGCGG

GAGTACCGGA

GCGGTGGCGA

AAGGCCAGCA

ACCACCGGGT

AAGGAACTTA

TTCACAGCCT

CAGCAGCCCT

GGCCGGCGGT

GTCAGGAGGG

GCGACTGGGT

TCGGCGGCAA

TA

INFORMATION FOR SEQ ID NO:17: SEQUENCE CHARACTERISTICS: WO 98/11205 PCT/NZ97/00112 LENGTH: 622 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17: CCAAAGCTCC TAGTGCCTCA TGAGTCTGCT GAGGATTGCA CAATTGGCGG GTTCGACGTG

CCCCGAGGCA

120

GACGATCCCA

180

CGACTATTGC

240

GTGGTGAGCT

300

GAATTGGTGG

360

GCCTTGTGCA

420

CGTTGTCTAA

480

CTGAAAATAG

540

GCCAATGCAG

600

AACATTGTTC

622 CCATGATCCT GGTTAATGCG CAAATTTTAA ACCGGAGAGG CGTTTGGGAT GGGGAGGAGA TGGTCCTGGC GGCGCTTATT ACTTGTCCGA GGGGACGGGA AAGCGCGTGA ATGCATGATA TGAATTTACA TTGGTGATGT GCCAGTGCAG CTTTAGGAAT CTTTAGGCCT TTCTCTTAGG AAAAAAAAAA AA

TGGGCAATTC

TACGAGGGAT

AGTTGTCCTG

CAGTGCTTCG

CTCACAATGC

GCTAATGTTC

ATCTCCAATG

GATCGTGAGC

AGAAAAATGA

AAAGAGACCC

TGGAAGGTGA

GTGCTGGCCT

AATGGGAACG

CAAAGAGAGA

TTGCGCACCT

TTTTTGAATA

ATCAATAGCA

TGGTTTATAT

AAAAGTGTGG

TCATGCCTAC

TGCCAATAGA

AGTTGGCGAA

GCCATTGGAG

TTAAGAAGGT

ATCAAATAGA

TCCTGAGGAG

AGGTACTGGC

INFORMATION FOR SEQ ID NO:18: SEQUENCE CHARACTERISTICS: LENGTH: 414 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

CACGCTCGAC

CATTGAACTC

120

ACCCCACCCA

180

CAATGCAGTC

240

CGGTGAACTG

300

TCTCCGGCAA

360

AGGAAGCCAA

414 SEQUENCE DESCRIPTION: GAATTCGGTA CCCCGGGTTC TCTCTCTCTC TCTCTCTCTC CATACAGACA AGTAGATACG AATCGCACTA GCGACGGTTC GGTGTGGCTG AGGCCGAAGA GTCCTACACC TTCCTGGTCG GTCCAAGCCC ATCGCCGTCT SEQ ID NO:18: GAAATCGATA AGCTTGGATC TCTCTCTCTC. TCCCCCACCC CGCACACAGA AGAAGAAAAG TGGCCGTCCT AACGACATGG GGCTCGAGAG GCTTCTGAGA GCGACCTCAA GGAGAACCTG CCGATGACAT CAAGCCTCGT

CAAAGCAACA

CCCCTTCCCA

ATGGGGGTTT

GCGTGGAGGG

CAGCAAGGTC

CGGATGCTCA

CTCT

INFORMATION FOR SEQ ID NO:19: SEQUENCE CHARACTERISTICS: LENGTH: 469 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear WO 98/11205 WO 9811205PCTNZ97OO1 12 (xi)

GAATTCGGCA

CTCATCTCCT

120

CTTCTCCGAA.

180

TCATAGGCCA

240

TGGCGGACAA

300

TAAGCAGCCG

360

GCCCCAAATC

420

AATACGGGGA

469 SEQUENCE DESCRI PTION: CGAGTGTCTC TCTCTCTCTC AGCAGTTCTA GGGGTTGTGT CAAACCCAAA GGTACTGCCT CATCCACTTG CTGGGCGGCG GCAGGGCCCG ATGTTTCGGA TGAGGCGGTC COGGACTGCT CAAGGCGGGA ATCCACTT-G CTTTTGGCGC GAGATGAGGA SEQ I D NO: 19: TCTCTCTGTA AACCACCATG TGCTCCTGCT AATTCTATGG TACCCCCGGA GCTGCCGGGC AGACCCCGCT GGCCAGGACC TCCGTCTCGG AGTCCACCCG TCACCACCCA CGACAAGGAC GCTACGGGTA TGCCGGTTTT AGATCACCAT GC-TCGAGCT

CTCTTCCTCA

AGGGCAAGAT

GCATGGCCGA

CTGGCCGCCA

GCGACCATCA

CTCGCTTCTC

GGCTTCGTAG

INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 341 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

CGGGCTCGTG

TCATCGGGAA

120

AGAAGTATGG

180

CCCCCGACGT

240

CCACCATCGC

300

GCCCGTTCTG

341 SEQUENCE DESCRIPTION: GCTCGGCTCC GGCGCAACGC CATGCTCATG ATGGGCGAGC CGGGATCTTC CACCTCCGCA GGCCCGCCAG GTCCTCCAGG GATCAGCTAC CTCACGTATG GCGGCAGATG CGGAAGCTGT SEQ ID CCTTCCCACC GGGCCCGAGG GGCCTCCCGG TCACCCACCG CGGCCTCGCG AGTCTGGCGA TGGGCTTCCT GCACATGGTT GCCGTGTCGT TCCACGACGG GATCTTCTCG AACCGGCCTG ACCGGGCCGA CATGGCCTTC GCGCACTACG GCGTGATGAA A INFORMATION FOR SEQ ID NO:21: SEQUENCE CHARACTERISTICS: LENGTH: 367 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

GAATTCGGCA

GAGGGGCCTC

120

CGCGAGTCTG

180

GGTTGCCGTG

240

CTCGAACCGG

300

CTTCGCGC.AC

360 SEQUENCE DESCRIPTION: CGAGCGGGCT CGTGGCTCGG CCGGTCATCG GGAACATGCT GCGAAGAAGT ATGGCGGGAT TCGTCCCCCG ACGTGGCCCG CCTGCCACCA TCGCGATCAG TACGGCCCGT TCTGGCGGCA SEQ ID NO:21: CTCCGGCGCA ACGCCCTTCC CATGATGGGC GAGCTCACCC CTTCCACCTC CGCATGGGCT CCAGGTCCTC CAGGTCCACG CTACCTCACG TATGACCGGG GATGCGGAAG CTGTGCGTGA

CACCGGGCCC

AC CGCGCCC T

TCCTCCACAT

ACGCCATCTT

CCGACATGC

TGAAAGCTCT

WO 98/11205 WO 9811205PCTINZ97/001 12 TCAGCGGAAG CGGGCTGAGT CGTGGGA 387 INFORMATION FOR SEQ ID NO:22: SEQUENCE CHARACTERISTICS: LENGTH: 443 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

CACGAGCTCG

CACTCCTTTC

120

AAAGCCCAAA

180

GAACCAAGCA

240

TGCTCTGATA

300

ACTGGCCGAG

360

CGCAAAGAAC

420

CTCCATCCTC

443 SEQUENCE DESCRIPTION: TGAGCCTTCC CGGAGACAAG TCAAGAAACC TAGTCATCCA CTCGTACAGA AGGAGAGAGA ATCACGACGG CCAGTGAAGA GCACTCCCCT TGGTCTTGAA TGCGGGCCTA TGGCTCCACT CCGGAAGCCC CCGTAACCCT TCTTGCACTC TCG SEQ ID NO:22: GCCATCTTAC TTCGCAACAA AGAAGCAGAG CATTGCAACT .GAGAGAGAAT AGAAGCATGA TGAAGAGTTC TTGTTCGCCA GGCCACCATC GAACTGGGGA TTCGCCTGCT CAGATTGCCT TGACCGGATC CTCCGGTTTC

ATTGCGTCCG

GCAAACAGCC

GTGCATGCAC

TGGAAATGAA

TCCTCGAAAT

CCCGTCTCTC

TCGCCAGCTA

INFORMATION FOR SEQ ID NO:23: SEQUENCE CHARACTERISTICS: LENGTH: 607 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

GAATTCGGCA

AACCGGTCCA

120

ACTTAGGTCA

180

AGAGAGAGAG

240

GGAGAGCCAG

300

TGATGCTCTT

360

GAAGGAGCTC

420

CGAAGGGCAG

480

TGGTGTCTTC

540

GATTTTGGCT

600

GCCGGTG

607 SEQUENCE DESCRIPTION: SEQ ID NO:23: CGAGCCAACC CTGGACCAGG TACTTTTGGC AGGCGGTCCA TTGCCCTTCA

AACCGGACCA

ACTGCAACAT

AGAGAGAGAG

ACCCAAGCCG

TACCAATATA

AGGGAAATAA

TTCTTGAACA

ACTGGCTACT

ATGGACATTA

TCACTGTCCT

TTCTTGATCA

AGAGAGAGAG

GGAGGCACCA

TTTTGGAGAC

CAGCAAAACA

TGCTTICTCAA

CTCTCCTCGC

ACAGAGAGAG

TATATACGTT

CAACATATTA

AGAGTTTGAA

GGAGGTTGGC

CAGCGTGTAC

TCCATGGAAC

GCTCATCAAA

CACCGCTCTT

CTATGAACTT

GCATCATGCC

CAATATTCCT

TCAATGGCCA

CACAAGTCTC

CCAAGAGAGC

ATAATGACAA

GCCAAGAACA

GCTCTTCCTG

GGCCTGCCGG

TGCTCATAGA

AAGCAGAGAG

CCGCCGGAGA

TCCTTCAGAG

CTGAGCCCAT

CATCAGCAGA

CCATGGAGAT

ATGACGGAAA

CAT CCAAAAA INFORMATION FOR SEQ ID NO:24: WO 98/11205 WO 9811205PCTINZ97/001 12 SEQUENCE CHARACTERISTICS: LENGTH: 421 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

GAATTCGGCA

AGAGAAGAGA

120

CCAAGTCTCG

180

CCCCATGGTC

240

GCCGGGCGCG

300

GGCACCCGTA

360

CACCCTCCGC

420

C

421 SEQUENCE DESCRIPTION: CGAGCCGTTT TATTTCCTCT GGAGAGGAGA GAATGGGTTC GACGAGGAGG CGAACCTCTT CTCAAGGCCG CCATCGAGCT TTCCTCTCCC CGGGGGAAGTI ATGCTCGACC GGATCTTCCG GACCTCCCCG ATGGCAAGGT SEQ ID NO:24: GATTTCCTTT GCTCGAGTCT GACCGGATCC GAGACCCAGA CGCCATGCAG CTGGCGAGCG CGACCTCCTC GAGATCATGG CGCGGCCCAG CTCCCGACCC GCTGCTGGCC AGCTACTCCG CGAGCGGCTC TACGGCTTAG

CGCGGAAGAG

TGACCCCGAC

CCITCCGTGCT

CCAAGGCCGG

AGAACCCCGA

TGCTCACGTG

CGCCGGTGTG

INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 760 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID GGAAGAAGCC GAGCAAACGA ATTGCAGACG CCATTGAAAA AAGACACGAA AGAGATCAAG,

AAGGAGCTTA

120

CGAAGCATTC

180

TATTGGAGAC

240

CAGCCAAACA

300

TGCTCCTCAA

360

CTCTCCTCGC

420

ATAGGGAGAA

480

TCGATTTCAG

540

ACCATGGAAC

600

TCAATGGTTC

660

TAAGGGTAGT

720

GCTGACCCGG

760

AGAAGCATCA

GGAAGTCGGC

CAGCGTCTAC

TCCATGGAAC

GCTCATCAAC

AACCGCCCTT

CTTCGAGATC

AGAAGGCCCT

GTACGACTTC

AAAGACAACA

TTCTCATTTC

CGGCACAGGT

TCAATGGCAG

CACAAGAGCC

CCAAGAGAGC

CTGATGACCA

GCCAAGAACA

GCTCTTCCCG

GGGCTGCCCG

GCCCTGCCGC

TTCTCAATCC

TAAGACAGAA

ATCAATGCTT

GATGCCATCC

CCAACGCAGA

TCTTGCAGAG

CAGAGCCCAT.

CATCGGCGGA

CCATGGAGAT

ATGACGGAAA

TCATCCAGAA

TCCTTGATCA

TTAATCGTTC

GATGGAAAAA

GATTTTGAGA

CCGACGGGAA

GCCTCAGCAG

CGATGCTCTC

TGAAGGGCAG

CGGCGTCTAC

GATCTTGGCC

GGCCGGCCTT

GCTCGTGCA.A

ATTTGAATAC

ATAGAAAGGA

TCTCCTTTCT

ACCCAACCAG

TACCAGTATA

AGGGAAATAA

TTCCTGAACA

ACCGGCTACT

ATGGCCATCA

GCCCACAAGA

GATGAGAAGA

AAATACATGC

AGGAAAGTAT

GGTGCGATCA

INFORMATION FOR SEQ ID NO:26:

.IW

WO 98/11205 PCT/NZ97/O01 12 SEQUENCE CHARACTERISTICS: LENGTH: 508 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: GAATTCGGTA CCCGGGTTCG AAATCGATAA

ACTAACCATC

120

ATATCGTGAA

180

AGGGACGGGC

240

GTTCGCGCTG

300

CTTCAAGAAC

360

GAAGGCAATC

420

TCAGACCAAA

480

GGTTCATTTG

508

TGCCTTTCTT

AGGAGTCCGT

TACGTCGGCA

GTTAGGCAGA

TTGGGCGTCA

AAGCAAGCCG

GAATCGTCGA

ATCTGGTTTG

CAT CTT CTTT

CGACGACAAT

ACT TCAT CGT

GCACGGTCTC

CTCTGCTCAT

ACGTGGTGAT

CGCCATTAAA

GGGGGGTC

SEQ ID NO:26: GCTTGGATCC AAAGAATTCG CTTCTGCTTC TCCTCCGTTT GGCCGAGAAG AGCAAGGTCC GGAAGCGAGT GCAAAAGCAG CGACCCCGTC ,A-AGGGCCAGC CGGTGATCTG TACGATCATG ATCGACAGTG GGGCACATGC GGAAGCTGGC AACGTTAAGG

GCACGAGATC

CCTCGTTTCG

TGATCAT CCC

CCCATCCCAC

TCCTCGAGAG

ACAGCTTCGT

TAA:ATGGCCGA

TTTGTTGGTT

INFORMATION FOR SEQ ID NO:27: Wi SEQUENCE CHARACTERISTICS: LENGTH: 495 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:27: GAATTCGGCA CGAGGTTAAT CGCACTGCAG CCTCAACACC ACCCACCTTC CTCCATCTCT

CTCCTCCCTT

120

AAGAGGAAGG

180

ATCCTCATCA

240

GAAGGTCATC

300

GGTGAGTCCG

360

AGAAAGGATT

420

GACATTAACG

480

ACCAGTCAAC

495

CTTCTTTCTC

TGGGGCAGCC

TCGGAGGCAC

AGGTCACTTT

ACAAGGACTT

TTGATTTTGT

CCGAGAGGCG

TACTG

TGACTTCAAT

TAAAGGGGCA

CCGTTTCATC

CTTTACCAGA

CGCTGATTTT

TAAATCTAGT

GATGAAGTCG

GGCAGCCGAC

CTGCGGGTCA

GCTGTGTTTT

GGAAAAGCAC

TCATCCAAGA

CTTGCTCCAG

CACCAATTTT

TCCATGCTTG

CTGCATCAAG

TGTCGAGACT

CCATCACTCA

TCCTGCATTT

AAGGCTTTGA

GGATGCCTGC

CGTTCAGTAT

CAATAAGAAG

ACTTGTCAAA

ACAATTGCCT

GAAAGGAGAC

CGTTGTTTAT

CAAACCTTGA

INFORMATION FOR SEQ ID NO:28: SEQUENCE CHARACTERISTICS: LENGTH: 472 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear WO 98/11205 WO 9811205PCT/NZ97/00112 (xi)

GAATTCGGCA

CCTCGTCGTT

120

GGGCCACCCC

180

GACGCTACTG

240

GAGGAGCCTC

300

CCACTTCCGG

360

TGGAAATGTC

420

TGCAATTGAG

472 SEQUENCE DESCRIPTION: CGAGCATAAG CTCTCCCGTA GGCGGCACTG GCTACCTCGG ACGTACGTCC TCCAGCGTCC CGCTTCAAGA GGCGTGGCGC GTCGACGCTG TGAGGCGGGT AGCCACAACA TCCTGATGCA AAGCGGTTTT TGCCGTCAGA CCGGGAAGGG TCACGTTCGA SEQ ID NO:28: ATCCTCACAT CACATGGCGA GCGGAGGTTC GTGAGGGCGA GGAGACCGGC CTCGACATTG CCAACTCGTC GAGGCCTCG-T CGATGTCGTC GTCTGTGCCA GCTCAAGCTC GTGGAGGCTA GTTCGGAATG GACCCGGCCC TGAGAAATGG. AGGTGAGAAA

AGAGCAAGGT

GCCTGGACCA

AGAAGCTCCA

TCTCAGACCT

TGTCGGGGGT

TCAAAGAAGC

TCATGGGTCA

AG

INFORMATION FOR SEQ ID NO:29: Wi SEQUENCE CHARACTERISTICS: LENGTH: 396 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

GAATTCGGCA

AGACGAAGGC

120

CGTCCGGTTA

180

AGGCCACCGT

240

GAGCGAAAGA

300

CTATTGTTGA

360

AGGATCCGCA

396 SEQUENCE DESCRIPTION: CGAGGAGGCA CCTCCTCGAA GAGAATGAGC GCGGCGGGCG CATCGCCTCG TGGCTCGTCA CCGCGATCCG AATGATCCAA TAGACTTCAA CTGTTCAAAG GGGTTGTGCA GGCGTTTTTC GGCAGAATTA CTTGATCCGG SEQ ID NO:29: ACGAAGAAGA AGAAGGACGA GTGCCGGGAA GGTCGTGTGC AGCTCCTCCT CCAGCGCGGC AAAAGACTGA ACATTTGCTT GAAACCTGCT GGAAGAGGGT AAACTGCCTC TCCCTTTTAT

CTGTAA

AGGACGAAGG

GTGACCGGGG

TACACCGTCA

GGACTTGATG

TCATTTGATC

CATGATGTCA

INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 592 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: GAATTCGGCA CGAGGTTGAA CCTCCCGTCC CGCTCCCGCA TACTCCACCA CCGCGTACAG 120 TGCCTCGGTT GGGCTGCCCG GGACCCTTCT 180 AGGGCCGTGG GAAGCGAAGA CGTCTCGATT 240 SEQ ID TCGGCTCTGC TCGGCTCGTC ACCCTCTTCG AAGATGAGCT CGGAGGGTGG GAAGGAGGAT GGGTTCCTCT CCCCCTACAA ATTCACCCGC AAGATCACGC ACTGTGGAGT GTGCTACGCA WO 98/11205

GATGTGGCTT

300

GAGATAGTTG

360

CATGTGGGGG

420

CTAGAAGTCC

480

GTGACAAAGG

540

CCAGAAAACT

592

GGACTAGGAA

GAATTGTGAA

TGGGAACTTA

AATGTGAAAA

GAGGATATTC

ACCCGATGGA

TGTGCAGGGA

ACAGGTTGGC

TGTCAATTCA

GTCGGTTATG

TAGTCACATT

TCTAGCAGCG

CACTCCAAG T

TCCAGTGTCC

TGCAGAGAGT

ACTT.TTGATG

GTCGTCCATG

CATTTGCTCT

A T C-CCTGGT

AACGCTTCAA

GCGAGTATTG

GAATTGATGC

AAAGGTATTG

GTGCTGGATC

PCT/NZ97/O01 12

'-CCAGGGCAC

A: GTTGGCGAT

:CAATGACAGG

.2GATGGTACA

CSTCAGGATT

INFORMATION FOR SEQ ID NO:31: SEQUENCE CHARACTERISTICS: LENGTH: 468 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

GAATTCGGCA

AAACAAATGG

120

TACCCTGCAC

180

GGCTTCCACA

240

GGCCCCGAGT

300

CTTCCTCCTT

360

AACTATATGG

420

CCTCCGGTGA

468 SEQUENCE DESCRIPTION: CGAGAACTCA TCTTGAAATG GTTCCGCCGG ATTCGAATCG AAAGCCACAT TGGCGCCATG TCTCCTTCGT CAACACCGAG TCACAAATGG AATGCTGAGC CGGACTTGGA TGCGATCCAA TCAGCCCCAT CAACGATCTT CTTGCATCAA TCTCGGATGG SEQ ID NO:31: TCATTGGAGT CATCATCCTC GCCACAAAGC CGCACGCCGT CTCAAGCTAG CAAAGCTCCT TTCAACCACC GGCGGCTCGC GACTTTCAGT TCCTGACAAT GACATCAAGA TGCTCTGCGA GTATCGAGCC TGGGCTCGAA TTTCATGACA CTCGTGAC

:AGTGAGAAG

T-7TGCATTCCC rCCrATCACAAG

CAGGGCTCGA

CCCCGATGGT

ATICGTCCAGG

CCCGAGCGTC

INFORMATION FOR SEQ ID NO:32: SEQUENCE CHARACTERISTICS: LENGTH: 405 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

CTTTACTCCG

GTCCATCTTC

120

AGCAAGCCCC

180

CGGAATTTTC

240

TTCGACCCGG

300

AGGCATTACA

360

ATCGGGCTGT

405 SEQUENCE DESCRIPTION: CCAAGAAGAT CCAATCGCAG ATCGGGAAGT CTCTTGGCAG TAACTCAGTG GTCTATGTGA CGAAATAGCT TTAGGTTTAG GTCAGTGAGC GGCTCGGAAC GGAGAGGGGG AAGATTGTGA CGGAGCGTTT TGGACTCACA SEQ ID NO:32: TTTTCGCAAT TGGCCCATTA AAGACCGGAG TTGCATTTCC GTCTTGGGAG CATCGCCTCT CCGATAGCCA GCAGCCATTC TCTTAGAGAA TTTGCCCGGT AATGGGCGCC TCAACATGAA ATGGATGGAA CTCCA

CACAAATGCG

TGGCTGGACA

GTGAACGAGT

TTGTGGGTGG

TGCTTTCTGG

GTGCTGGCTC

tl I

IY

WO 98/11205 PCTNZ97/0O112 INFORMATION FOR SEQ ID NO:33: i) SEQUENCE CHARACTERISTICS: LENGTH: 380 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

GGCAAACACG

TCGAGAGCTT

120

AGGAGCTCAC

180

TCCCGACCAT

240

ACGAGGAGCT

300

TCCCCAACGA

360

TCGAGGAGAA

380 SEQUENCE DESCRIPTION: CCCGTTTTCG TTTTACTAAG GTCGAGCAGT GGCATTCAGT AAGCATTGGC GACATCTTCG CGACCTCGAG GACATAGCGT CAGGAAGGCT GCCACCGACT CCTGATTGAG CGTGTAAAGA

GGACAAGCAT

SEQ ID NO:33: AGAAGATGGT GAGCGTTGTG CGATCCCGCA GGAGTATGTG AGGAGGAGAA GAAGCATGAG

CTAAAGACCC'CGTGGTGAGG

GGGGCGTCAT GCACCTCGTC AGGCTGGCGA GGTGTTCTTC

GCTGGTAGAG

AGGCCGAAGG

GGCCCTCAGG

GAGAGGTGCC

AACCATGGGA

AACCTCCCGA

INFORMATION FOR SEQ ID NO:34: Ci) SEQUENCE CHARACTERISTICS: LENGTH: 305 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

TTGTACCCGA

CGAGGCCGCC

120

CGTCGTGGTC

180

AGGAAGCGGA

240

TCGGAGATGG

300

TGTTT

305 SEQUENCE DESCRIPTION: AGATCTCCGG GACCGTTCGA GCCGGAGGCC GGGGAGAAGC GGCGGCGGCG GCGTGGTGGA CATGGCTGGG GGATCGATCG AGAGATGGAA ATGAAAGAGA SEQ ID NO:34: CGGCGACATC GCCGTCGGCC GGGAACCCGT TGGAGTAGCC GCCGTAGCCG GAGAAGGCGC CCTCATCGCC GTCCATGCTG AAGGCGTCGA ACCGATCCGA TCGGCCGGAG GATTTCGAGA GAGAGAGAGA GAGATCCGGT GGACTGGTGG INFORMATION FOR SEQ ID Ci) SEQUENCE CHARACTERISTICS: LENGTH: 693 base pairs TYPE: nucleic acid CC) STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID GAATTCGGCA.CGAGCTAAGA GAGGAGAGGA GAGGAGCAAG ATGGCACTAG CAGGAGCTGC ACTGTCAGGA ACCGTGGTGA GCTCCCCCTT TGTGAGGATG CAGCCTGTGA ACAGACTCAG 120 WO 98/11205 WO 9811205PCT/NZ97/001 12

GGCATTCCCC

180

TGCCATGGCC

240

CCCCGACGAT

300

CTGCCGTGCC

360

GAGCGACGGC

420

CGCCTACCCT

480

AAGCTCTCCT

540

TCTCCCCCCT

600

GGGATGATTT

660

TGAGGAAATA

693

AATGTGGGTC

GCTTACAAGG

GTTTACATCT

GGCTCTTGCT

AGCTTCCTGG

AAGTCTGAGG

ATATTTGCTT

TCACTACATG

GATGTTATTC

AAACTCATGC

AGGCCCTGTT

TCACCCTGCT

TGGACTACGC

CCTCCTGCGC

ATGATGATCA

TCACCATTGA

TTGCATAAAT

TTTGTTAGTT

TGAGTCTAAT

TCTAAAAAAA

TGGTGTCAALC

CACCCCTGAA

CGAGGAGCAA

GGGCAAGGTC

GATTGAGGAA

GACCCACAAG

CAGTCTCACT

CCTTTAGTCT

GTAATGGCTT

AAA

TCTGGCCGTG

GGCAAAGTCG

GGC-ATCGACT

GTGGCGGGGA

GGTTGGGTCC

GAAGAGGAGC

CTACGCAACT

CTTCCTTTTT

TTCTTTTTCC

GCAGAGTGAC

AACTCGACGT

TGCCCTACTC

GCGTCGACCA

TCACTTGTGT

TCACTGCTTG

TTCTCCACTC

TACTGTACGA

TATTTCTGTA

INFORMATION FOR SEQ, ID NO:36: SEQUENCE CHARACTERISTICS: LENGTH: 418 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

AGGACTTTAT

TCCAATAATC

120

ATGCCTTAGT

180

TGTGAAAAAC

240

GATATGATGA

300

TCCTCGGCAG

360

ACATCCCGCA

418 SEQUENCE DESCRIPTION: SEQ ID NO:36: TATAAGCATT GTAAAAAGAG TCAAACTAAT ACATCGCAAG TACAAAAAGA AAAAAGTTTG ATGCATTGAG ATGGTAACTG TTGAAAAATT AACCAACTAT TAAAATTAAT GATGATGAAT TATATAGACT TAAAATTGAC TCAGAAGACA TTC!TTTTCTT ATTCGGTCTA AACAGGCAAA TGGTGTCAA.A CGGGAAGTCG TGACTACCGG GCGGGCGATG .ATGCGGATCC GGGGGCCGGG CGGACCGGTC CACGTTTGGT GCGGTGACAA CAGGCAGCCC

AATTGGGTTA

CTTAATTCAA

ATGGATTATG

CTTATTTTAT

GCAAAACTCT

TCGCTGGAGA

AACCTGGA

INFORMATION FOR SEQ, ID NO:37: SEQUENCE CHARACTERISTICS: LENGTH: 777 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:37: GAATTCGGCA CGAGCATACA ACTACACTGC GACGCCGCCG CAGAACGCGA GCGTGCCGAC CATGAACGGC ACCAAGGTCT ACCGGTTGCC GTATAACGCT ACGGTCCAGC TCGTTTTACA 120 GGACACCGGG ATAATCGCGC CGGAGACCCA CCCCATCCAT CTGCACGGAT TCAACTTCTT 180 CGGTGTGGGC AAAGGAGTGG GGAATTATGA CCCAAAGAAG GATCCCAAGA AGTTCAATCT 240 WO 98/1 1205

GGTTGACCCA

300

ATTCACAGCA

360

TTGGGGACTG

420

TCCACCTCCA

480

ACTAATGACA

540

ATAAGAAAGA

600

CCAAAGAGAC

660

ACTCCGACAC

720

AGTGTAATTT

777

GTGGAGAGGA

GACAATCCAG

AAGATGGCAT

AGTGATCTTC

CCAAGTTAGT

TGAGGAGAGA

CCTTGAGATC

TGCTACAATA

GTTTTTTGGC

ACACCATTGG

GAGTTTGGTT

TCTTGGTGGA

CAAAATGTTG

GGAATCTTCT

AGCCATAGAA

ACGACATCCC

AATTAAGGAA

AAGCTCATCA

AATCCCATCT

CCTGCACTGC

CAATGGGAAG

ATCATTTGAT

CTTTGAAAAA

GATTTGACCA

GCAATTGTTT

GACAAGGAAT

CATGAATCAC

GGTGGATGGA

CATCTGGAAG

GGGCCTAAAG

CATGAGGACG

GAAGAAGAAG

AGAAGAGAGA

CTAGAGTAAT

TTGGTTTTTT

AT GGAAAAAA PCTNZ97/0O1 12

TAGCCATCAG

TGCACACAAC

AGACCCTGCT

ACAAGCGATT

AGCAAGAAGA

GGGCAATAAA

AGAAGGATTT

TCATTGGAGG

AAAAAA

INFORMATION FOR SEQ I0 NO:38: Ci) SEQUENCE CHARACTERISTICS: LENGTH: 344 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

ATATGTTCAG

ATACGTTGAA

120

TCGATGTTCA

180

ACTACTACAT

240

TACACTACAC

300

AAAAACATTG

344 SEQUENCE DESCRI PTION: AATTTCAAAT GTGGGAATGT GCTAGTCGAG GTTGAAGGAT CGTGGGCCAA TCCATGGCTG TGTCGCATCC ACCCGGTTCA CAACTCGCTT ACCCCAGTTT GTCCATGAAG CAAGCAAGAA SEQ ID NO: 38: CAACCTCCTT GAACTTCAGA ATTCAGGGCC CTCACACCGT CCAGAACATG TATGATTCAA TCTTAGTGAC CTTAAATCAG CCTCCAAAGG CCAAGACGGT TCTCAATGCA ACTGCAGTGC CCGGGCCACT ACCAGCTGGT CCAACTTACC CAATCAGGTG GAAC INFORMATION FOR SEQ ID NO:39: SEQUENCE CHARACTERISTICS: LENGTH: 341 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

GCCGCAACTG

GGGAGCTCTC

120

TGATTTTGTC

180

GGTGAACGGG

240

CAATGTCGTC

300

GAGATCTGGT

341 SEQUENCE DESCRIPTION: CAATTCTCTT CGTAAAACAT CTCCTCTTCT CTGTGGCGGT GTTCAAGCGA CCAAGGTGAA CAATTCCCGG GTCCGACTTT AACAAAGCTC GCTACAACGT TGGGCTGATG GGGCGGAATT SEQ ID NO:39: GACGGCTGTC GGCAAAACCT CTTTCCTCTT GACATTGGCA GATGCAAAAG TTTACTACCA GAGGCTGTGC ACGACCCACA ACACCATCAC GGAAGTTAAC GACGGCGACA CCCTCGTTGT CACCATTCAC TGGCACGGCG TCCGGCAGGT TGTGACTCAA T 1 1

I

WO 98/11205 PCTINZ97/00112 INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 358 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

GAATTCGGCA

CAGAATTCAG

120

CATGTATGAT

180

TCAGCCTCCA

240

TGCAACTGCA

300

TGGTCCAACT

358 SEQUENCE DESCRIPTION: CGAGATATGT TCAGAATTTC GGCCATACGT TGAAGCTAGT TCAATCGATG TTCACGTGGG AAGGACTACT ACATTGTCGC GTGCTACACT ACACCAACTC TACCAAAAAC ATTGGTCCAT SEQ ID AAATGTGGGA ATGTCAACCT CGAGGTTGAA GGATCTCACA CCAATCCATG GCTGTCTTAG ATCCACCCGG- TTCACCAAGA GCTTACCCCA GTTTCCGGGC GAAGCAAGCA AGAACAATCA

CCTTGAACTT

CCGTCCAGAA

TGACCTTAAA

CGGTTCTCAA

CACTACCAGC

GGTGGAAC

INFORMATION FOR SEQ ID NO:41: Wi) SEQUENCE CHARACTERISTICS: LENGTH: 409 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

ATCAAGAGTT

TGCTGATCGG

120

TTCCTCTTGG

180

TACTACCATG

240

ACCATCACGG

300

CTCGTTGTCA

360

CGGCAGGTGA

409 SEQUENCE DESCRIPTION: TGAGTCTAAA CCTTGTCTAA CCGCAGCTGC ATTCTCTTCG GAGCTCTCCT CCTCTTCTCT ATTTTGTCGT TCAAGCGACC TGAACGGGCA ATTCCCGGGT ATGTCGTCAA CAAAGCTCGC GATCTGGTTG GGCTGATGGG SEQ ID NO:41: TCCTCTCTCG CATAGTCATT TAAAACATGA CGGCTGTCGG GTGGCGGTGA CATTGGCAGA AAGGTGAAGA GGCTGTGCAC CCGACTTTGG AAGTTAACGA TACAACGTCA CCATTCACTG GCGGAATTTG TGACTCAAT

TGGAGACGAA

CAAAACCTCT

TGCAAAAGTT

GACCCACAAC

CGGCGACACC

GCACGGCGTC

INFORMATION FOR SEQ ID NO:42: i) SEQUENCE CHARACTERISTICS: LENGTH: 515 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:42: CTCTCTCTCT CTCTCTCTCT GTGTGTTCAT TCTCGTTGAG CTCGTGGTCG CCTCCCGCCA TGGATCCGCA CAAGTACCGT CCATCCAGTG CTTTCAACAC TTCTTTCTGG ACTACGAACT 120 .4) WO 98/11205

CTGGTGCTCC

180

TTCTTGAGGA

240

AGCGTGTGGT

300

TTTCCCAGCT

360

TCCGTTTCTC

420

GTTTTGCTGT

480

CTGTCTTCTT

515

TGTCTGGAAC

TTATCACCTC

GCATGCCAGA

TACCTGTGCT

CACTGTCATC

GAAGTTCTAC

TGTCCGTAAT

AATAACTCTT

GTGGAGAAAC

GGAGCCAGTG

GATTTCCTTC

CACGAAAGGG

ACAAGAGAGG

GGGATAAATT

CGT TGACTS T TTGCCAACT T

CAAAGGGATT

GGGCACCAGG

GCAGCCCT GA

GTAACTTTGA

CCCCG

TGGAAGCAGA

TGATAGGGAG

C-LTTGAGGTC

AGTTCAAACA

AACCC7TGAGG

TCTGGTGGGA

PCT/NZ97/OO1 12

CGTCCAATTC

AGGATTCCAG

AC'.TCATGACA

CCCGTGATTG

GACCCTCGAG

AACAATTTCC

INFORMATION FOR SEQ ID NO:43: SEQUENCE CHARACTERISTICS: LENGTH: 471 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

GAATTCGGCA

ATTTGCGGCG

120

GTAAGCCAGG

180

GCTGAGAAGA

240

GATGTGAAGA

300

CACGGGGCCA

360

TTCCCCGTCA

420

ACTGGTGGAC

471 SEQUENCE DESCRI PTION: CGAGGCTCCC TCTCGTACTG ATCCATTTCT CGATTCAAGG AGTACAAGAA GGCTGTCGAG GCTGCGCTCC GCTCATGCTC CGAAGACCGG AGGCCCGTTC ACAGCGGGCT CGACGTTGCC TCACTTATGC TGATTTCTAC CTGAAGTTGC TTTTCACCCG SEQ ID NO:43: CCATACTCCT GGGACGGGAT GGAAGAATCA TGGGGAAGTC AAATGCAAGA AGAAGTTGAG CGCATCGCGT GGCACTCCGC GGGACCATGA AGCACGCCGC GATCAGGTCT TGCAGCCGAT CAGCTGGCTG GCGTCGTTGC GAAGAGAGGC AAACCACAAC

TCGGATAGGG

CTACCCGACC

AGGCCTCATC

CGGTACCTTC

GGAGCTCAGC

CAAGGATCAG

TGTGGAAGTT

C

INFORMATION FOR SEQ ID NO:44: SEQUENCE CHARACTERISTICS: LENGTH: 487 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

GAATTCGGCA

TCAGTTTCGT

120

CGTCCAGCGC

180

ATGACTCATC

240

TTGAGAAACT

300

GAGCCAGCGC

360 SEQUENCE DESCRIPTION: CGAGCTCCCA CTTCTGTCTC GCTCTCTTCG TCATCTCTGC TTACGATTCC AGCTTTTGGA GCTGACTGTT GGAACTAGAG TGCCAACTTC GAGAGAGAGA GAAAGGGTTC TTCGAGGTCA SEQ ID NO:44: GCCACCATTA CTAGCTTCAA CTCTTGCCAT GGATCCGTAC CAACCAACTA CGGTGCTCCC GTCCGATTCT CCTGGAGGAC GGATTCCTGA GCGGGTGGTC CCCACGACAT CTCTCACTTG

AGCCCAGATC

AAGTATCGCC

GTCTGGAACA

TACCATCTGA

CATGCACGGG

ACCTGTGCTG

WO 98/11205 WO 9811205PCT/NZ97/O01 12 ATTTCCTCCG GGCTCCTGGA GTCCAGACGC CCGTAATCGT CCGTTTCTCC A CCGTCATCC 420 ACGAGCGCGG CAGCCCGAAC CTCAGGGACC CTCGTGGTTT TGCAGTGAAG T1,CTACACCA 480

GAGAGGG

487 INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 684 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

GAATTCCTGC

GCGCGCCTGC

120

TTGTTGGACG

180

CCCAAGGAAG

240

GTCTGTTTTG

300

GAGGTGATGC

360

CCAGGGCAGA

420

AAAGAAGCAG

480

GCTCTGCGAA

540

CACTCCATCG

600

GACATGGCTC

660

ATGCGAATCT

684 SEQUENCE DESCRIPTION: SEQ ID AGCCCGGGGG ATCCACTAGT TCTAGAGCGG CCGCCACCGC GGTGGAGCTC

AGGTCGACAC

CCATGGAAGC

GACTGGCTCT

ACGCCAACGT

AAGGGAAACC

TCGAAGCCGC

CGCGGCTTCA

CATCGCCACA

AGCGGGAGAT

TCCACGGCGG

CTTTGGCAGC

TAG TGOAT CC

TCTCCGGAAA.

CGTCAACGGC

GCTGGGCGTG

GGAGTTCGTA

GGCCGTCATG

CGAGAAAGAC

GTGGTTGGGG

CAATTCCGTC

CAACTTCCAG

CGTC

AAAGAATTCG

GCCGGGATTC

ACAGCGGTGG

CTGGCTGAGA

GATCCGTTAA

GAGTTCCTCC

CCGTTGAGCA

CCTCCGATCG

AACGACAAT C

GGAACACCCA

GCACGAGGCC

TGGAACCOTT

GATCCCCGT

TTCTGTCTGC

CCCACCAGTT

TCGACGGTAG

AACCGAAACA

AAGTCATCCG

CGTTAATCGA

TCGGAGTTTC

CGACGGCCAC

TAAACTGCAG

GGCCGCGTCC

GCTCTTCTGC

GAAGCACCAC

CGACTACGTG

AGACCGCTAC

CGCTGCTACT

TGTCTCCAGG

CATGGACAAC

INFORMATION FOR SEQ ID NO:46: SEQUENCE CHARACTERISTICS: LENGTH: 418 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

GAATTCGGCA

GAACTCCTGG

120

TATGATTTGA

180

TGGTCATAAA

240

CATTGGATTT

300

AAAGTGGCGG

360 SEQUENCE DESCRIPTION: CGAGGACAAG GTCATAGGCC CCCATTCTGA AATAAATAAT GTCCTCGGAT CTTTTTGTTG GCTTGATTTT GTTTTTCTTT GCCAGAAATA TGTAAGGGTG ATCATTTGGG TAGCATGCAG SEQ ID NO:46: CTCTCTTCAA ATGCTTGGAT CTTCCAAGAT CGCCTTTATA ATGCAGTTGT TTACCGATCT CTTTTGTTTT ATACTOCTOG GCAGATCATT TGGGTGATCT ATCAGTTGGG TGATCGTGTA

GGGTGGAAAG

CAACGACTGC

GGAATTTGAT

ATTTGCATCC

GAAACATGTA

CTGCTTTCAC

11 WO 98/11205 WO 9811205PCTNZ97OO1 12 TATTACTTAC ATATTTAAAG ATCGGGAATA AAAACATGAT TTTAATTGAA. A.;AAAAAA 418 INFORMATION FOR SEQ ID NO:47: SEQUENCE CHARACTERISTICS: LENGTH: 479 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

GATATCCCAA

CGAGCAAGGA

120

TTAAAAGCAC

180

TTCGAGCAGC

240

CGTATTTGGG

300

CTGCCGTTGC

360

GGGTCGAGGA

420

GTGTCACTAC

479 SEQUENCE DESCRIPTION: CGACCGAAAA CCTGTATTTT AGAAAATATG GTTGCAGCAG TGGGCTGTGC ACGGACTTCG CAAGGCCATG GAAGGAAGTC AGCCAAGGAG ATTTCCATTG TCGAAGATCG CAAGTGAAAG GAGTTCAAAC TGGGTTCTCA TGGTTTCGGA GCCACTTCTC SEQ ID NO:47: CAGGGCGCCA TGGGGATCCG CAGAAATTAC GCAGGCCAAT GCTCGTCTGG CA'GCGATCCA ACTTTGAAGA AGTGAAAGCG AAGGGAAATC TCTGACAATC TGAA-ATTGGA TGCTGCGGCT CCCAGATGAC CAAGGGGACG ACAGGAGAAC GAACCAGGGA GA ATTCGGCA

GAAGTTCAAG

'CTGAACTGGG

ATGGTGGATT

TC-AGACGTTG

GCCAAATCTA

GATACCTATG

GCCGAGCTT

INFORMATION FOR SEQ ID NO:48: SEQUENCE CHARACTERISTICS: LENGTH: 1785 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

TATCGATAAG

CGCCACCGCG

120

CACGAGGTTG

180

ATATGGCAAA

240

CGATCTCGCC

300

CGTGGTGTTC

360

TCACTGGAGA

420

GCACTACAGA

480

CGCCGAGTCT

540

TATTATGTAT

600

CAAGCTCAAG

660

TGGGGATTTC

720 SEQUENCE DESCRIPTION: SEQ ID NO:48: CTTGATATCG AATTCCTGCA GCCCGGGGGA

GTGGAGCTCG

CAGGTCGGGG

ATCTTTCTGC

AAGGAGGTCC

GATATCTTCA

AAGATGCGCA

TTCGCGTGGG

TCCACCTCGG

AGGATGATGT

GCCCTCAACG

ATTCCCATTC

CGCGCCTGCA

ATGATTTGAA

TCAAGATGGG

TGCACACCCA

CGGGCAAGGG

GGATCATGAC

AAGACGAGAT

GCATTGTCAT

TCGACAGGAG

GAGAGCGAAG

TTAGGCCCTT

GGTCGACACT

TCACAGAAAC

CCAGAGGAAT

GGGCGTCGAG

GCAGGACATG

TGTGCCTTTC

CAGCCGCGTG

CCGTAGGCGC

ATTCGAATCC

TCGATTGGCC

CCTCAGAGGT

T.CCACTAGTT.. CTAGAGCGGC AGTGGATCCA AAGAATTCGG CTCAGCGATT TTGCCAAGAA CTTGTGGTAG TTTCATCTCC TTTGGGTCTC GAACCCGGAA GTGTTCACCG TCTATGGAGA TTTACGAATA AAGTTGTCCA GTCGCGGATG TGAAATCCCG GTCCAGCTCA TGATGTATAA GAGGACGACC CGCTTTTCCT CAGAGCTTTG AGTACAATTA TATCTCAGAA TCTGCAATGA WO 98111205

GATTAAAGAG

780

CAACAGTACC

840

TAGATGCTCA

900

TCAACGTTGC

960

TGAACCACCA

1020

GCGTGCAGAT

1080

AAACCCTTCG

1140

CCAAGCTCGG

1200

TGGCCAACAA

1260

AGGAGGAGAA

1320

GAGGAGGAGC

1380

ACTTGTTCAG

1440

GAAGGGCGGG

1500

AGCTTCTGCT

1560

AAACACTCCA

1620

TAGCACTTCA

1680

AATAAAAGTT

1740

GGGGAATTTT

1785

AAACGGCTCT

AAGACTAGTA

GGACAAGGGA

AGCAATTGAG

GGACATTCAG

AACGGAACCA

TCTCCGCATG

GGGCTACGAT

CCCCGCCAAC

GCACACCGAA

TGCCCGGGAA

AACTTCCACC

CAGTTCAGCC

TAATCCCAAC

TCTATCATGA

AAAGTTTGCT

TGCATAAATT

ACTGCTAAAA

CTCTTTTCAA

CCAACACCGG

lGAGATCAATG

ACAACGCTGT

AGCAAGGTGC

GACACGACAA

GCGATCCCCT

ATTCCGGCAG

TGGAAGAACC

GCCAATGGCA

TCATTCTGGC

TTCTGCCGCC

TTCACATTCT

TTGTCAGTGA

CTGTGTGTGC

AGGATTTCAA

AAATGATATT

GGACTACTTC

GGGAGCTCAA

AGGATAATGT

GGTCGATGGA

GCGCAGAGCT

GGTTGCCCTA

TGCTCGTCCC

AGAGCAAGAT

CCGAGGAGTT

ACGACTTCAA

GCTGCCTCTC

GCCCGGGCAG

CAACCATTCT

CTGGTATATA

GTGTCCACTG

TAACAGACAC

TCAATATACT

GTGGAAGAGC

GTGTGCA.ATG

TTTGTACATC

ATGGGGAATA

GGACGCTGTT

CCTTCAGGCG

CCACATGAAT

CCTGGTGAAC

CC-GCCCCGAG

ATTCCTGCCT

CTCGCACTCT

AGCAAAGTGG

CTCATCG-TCG

AATGCGCGCA

TCGAGTCTAC

CGTCAATTAT

ATTTTGACTC

PCTINZ97/001 12 '3CAAGAAGCT

GACCATATTT

GT TGAGAACA

GCGGAGCTGG

CTTGGACCAG

GTTGTGAAGG

fCT7CCACGACG

GCCTGGTGGT

C3-GTTCTTCG

T-CGGTGTGGG

CCATCGGAAG

ATGT CACTGA

CCAAGCCCAT

CCTGAACAAA.

TAAGAGCTCA

GTCATGTTTC

TCCACCAATT

INFORMATION FOR SEQ ID NO:49: SEQUENCE CHARACTERISTICS: LENGTH: 475 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

GAATTCGGCA

GTCCTCGTCC

120

TAGGAACCTG

180

ATTTTCCAGC

240

ATTCACTGTG

300

CGAGGCGCTC

360

GAAAATCTTC

420

GAGCCTTCGC

475 SEQUENCE DESCRIPTION: CGAGATTTCC ATGGACGATT TCGTTTTCCT TGTTCTTCCT CCGCCAGGAC CCCCGGCATG GGCGCGTTCG AGACCTCAGT TGGCTCGGTT CCCGCCCTCT GTACAGAAGG GCTCCGTCTT AGTAGCAACC AGCACAACAT AGGAATCTGG TTAAAGAAGC SEQ ID NO:49: CCGTTTGGCT TCAATTCGTT CCGACTTTTT CTCTGGAAGC GCCGATCGTA GGGAACGTCC GAAGAAATTC CATGAGAGAT GCTGATGATC ACCGACCGCG CGCTGACCGC CCGCCCGCCC CACTTCGGCT GAATACGGCC CCTGAGACTT CGGCGATGAA

TCCTCTGGCT

TATGGCGTAA

TTCAGATTGG

ACGGTCCAAT

AGCTTGCCCA

TCGGGATGCA

CGCTGTGGCG

GGCTT

INFORMATION FOR SEQ ID WO 98/11205 WO 9811205PCTNZ97OO1 12 SEQUENCE CHARACTERISTICS: LENGTH: 801 base pairs TYPE: nucleic acid STRANDEDNESS: sinqle TOPOLOGY: linear (xi)

GCTCCACCGA

GGCAATTCGA

120

CGTATGTTGT

180

GATTACGCCA

240

GGTGGCGGCC

300

CAGGCCTGAG

360

GCCATTCGGT

420

GTCTATGTTG

480

AGAAGACATA

540

GGCCATTGCT

600

AATTGATCTG

660

CTCCTTTTCC

720

CTGAAGCCCA

780

AATTGAGTAT

801 SEQUENCE DESCRIPTION: SEQ ID CGGTGGACGG TCCGCTACTC AGTAACTGAG

TTTAGCTCAC

GTGGAATTGT

AGCGCGCAAT

GCTCTAGAAC

AGATTTCTTG

GCAGGGCGCA

GGACACCTGC

GATCTCACAG

ATTCCTCGAT

ATAGTAAGTT

ATAGTCAACA

ACTTCTAGCA

TTCTCTGTAG

TCATTAGGCA

GAGCGGATAA

TAACCCTCAC

TAGTGGATCC

AGGAAGATGT

GGATCTGCCC

TTCATCATTT

AGAATCCAGG

TGCCTGATCA

TGAATTTTGT

TGCAGCTTTC

AGCAATAACT

CCCCAGGCT T

CAATTTCACA

TAAAGGGAAC

.AAAGAATTCG

TGATATTAAG

TGGTGCACA.A

CGTATGGGCA

GCTTGTTACT

TCTCTACAAG

TTTGATACAA

TTTCTCTGAA

GTATATTTTA

TGGGATCCCC CGGGCTGACA TACACTTTAT GCTTCCGGCT CAGGAAACAG CTATGACCAT AAAAGCTGGA GCTCCACCGC GCACGAGACC CAGTGACCTT GGCCATGATT. ACAGOCTACT TTGGGTATTA ATTTAGTTCA CCTCCTGAGG GAATGAAGGC TTCATGGCCA AGCCTGTGCA CGACAGCCAC TCAATTGATC AACGAAATAA CGTGCAGTTT GCGCATGCAG CTTTCTTTCT GAACAAATAC CTATTCCTCA INFORMATION FOR SEQ ID NO:51: SEQUENCE CHARACTERISTICS: LENGTH: 744 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

GGGCCCCCCT

AAGGACGCTG

120

GAGTACCCGG

180

TCTACCATGT

240

TTGGTGGATG

300

CACATTTCAG

360

GCTGTGAAAC

420

ATGAAGTGGA

480 SEQUENCE DESCRIPTION: TCGAGGTGGA CACTAGTGGA TGCTTGAAGG CTCCCAGCCA CCATCGATCA GAGATTCAAC TGATGAACAA GATTTTGGAT TGGGAGGAGG TATTGGGTCG GAATCAACTT CGACTTGTCC ATGTGGGTGG AGACATGTTT TTCTGCATGA TTGGAGCGAT SEQ ID NO:51: TCCAAAGAAT TCGGCACGAG TTCACCAAAG CCCATGGAAT AAGATTTTCA ACAGGGCTAT ACTTACGAGG GTTTTAAGGA ACTCTCAATC TCATAGTGTC CATGTGCTGG CCGATGCTCC GATAGTGTAC CAAGTGGCCA GATCATTGCA GGAAGCTTTT

GTTTTATCTG

GAATGCGTTC

GTCTGAGAAT

GGTTCAGGAG

TAGGTATCCC

TCACTACCCA

AG C TATTTT T

GAAGAATTGT

WO 98/11205 PCT/NZ97/00112 CACAAGGCGT TGCCAGAGAA GGGGAAGGTG 540 GCAGAGACAT CTCCTTATGC TCGTCAGGGA 600 AACCCAGGGG GCAAGGAACG CACAGAGCAA 660 TTTGCAGGTG GTGTTGAACC TGTATGTTGT 720 CAGCCCGGGG GATCCACTAG TTCT 744 ATTGCGGTGG ACACCATTCT TTTCATACAG ATTTACTGAT GAATTTCAAG ATTTAGCTAA GTCAATGGAA TGTGGGTAAT

CCCAGTGGCT

GTTGGCATAC

GGAGACGGGA

GGAATTCCTG

INFORMATION FOR SEQ ID NO:52: SEQUENCE CHARACTERISTICS: LENGTH: 426 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

GTGGCCCTGG

GGTTTCTCTC

120

AGCTCAAGAA

180

CATTGCTGGG

240

TTGTCTATCT

300

AGGCGTTTCT

360

CGTACATCGC

420

TGGAGC

426 SEQUENCE DESCRIPTION: AAGTAGTGTG CGCGACATGG TTGGCTTGCT CTCTACATTG GAGGCGCCTC CCGCCGGGCC AGCGATGCCT CACGTTACTC CAAACTGGGG ACGTCCGACA GAAGACTTTG GATATAAACT CTACGATTCT CAGGACATGG SEQ ID NO.:52: ATTCCTTGAA TTTGAACGAG GATTTCGTTA TGTTTTGAGA CATCGGGATG GCCAGTGGTG TCTACAACAT GTATAAGAAA TGGTTGTGGC CTCCACGCCC TCTCCAACCG GCCGGGAAAT TGTGGGCAGC GTATGGAGGA

TTTATGTTGT

TCGAACTTGA

GGAAGTCTGC

TATGGCCCCG

GCTGCAGCTA

GCAGGAGCCA

CGGTGGAAGA

INFORMATION FOR SEQ ID NO:53: SEQUENCE CHARACTERISTICS: LENGTH: 562 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

CAGTTCGAAA

ACACTAGTGG

120

CAGGATTTCT

180

AGAGGTGAAG

240

AGTGGGACAC

300

CGTGTACCCT

360

CTGGAACCTC

420

CATTAACGCC

480 SEQUENCE DESCRIPTION: TTAACCTCAC TAAAGGGAAC ATCCAAAGAA TTCGGCACGA TCTTGTCCAA ACAGGTTTAA GCTCAGACAA CCCAAGCAGA AAAAGTCTTT TGCAGAGCGA CGTGAGCCCG AGCCAATGAA ATGACTACTT CTGCCGATGA AAGAACACCA TGGAGATTGG SEQ ID NO:53: AAAAGCTGGA GTTCGCGCGC GCTTTGAGGC AACCTACATT GGAAATGGCA GGCACAAGTG GGAGCCGGTT AAGGTTGTCC TGCCCTCTAT CACTATATAT GGAGCTCCGC GAAGTGACTG GGGTCAATTT CTGGGCCTCC GGTGTACACT GGTTACTCGC

CTGCAGGTCG

CATTGAATCC

TTGCTGCAGC

GCCATCAAGA

TGGAAACGAG

CCAAGCATCC

TGCTGAAGCT

TTCTCAGCAC

WO 98/11205 PCT/NZ97/00112 AGCCCTTGCA TTGCCCGATG ATGGAAAGAT TCTAGCCATG GACATCAACA GAGAGAACTA 540 TGATATCGGA TTGCCTATAA TT 562 INFORMATION FOR SEQ ID NO:54: SEQUENCE CHARACTERISTICS: LENGTH: 1074 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

TCGTGCCGCT

GCACGCTGAG

120

GAATGGGAAA

180

GCTTCAAAAC

240

TCGAGAGGAC

300

GGAGATGATG

360

TGCTAAAAAG

420

CGCTCTTCCT

480 CT GGCCCTG C 540

GTTGGACTAT

600

CGTGGACGCA

660

CGTGGGGGGC

720

TCCCCACAAC

780

CAACTCCATG

840

TGTCACTGTT

900

TATATGATAA

960

TTTCTGTTTA

1020

GCCGCTTGCA

1074 SEQUENCE DESCRIPTION: SEQ, ID NO:54: CGATCCTCAC AGGCCCTTTT TATTTCCCTG

AATGGCAACG

CCAGGTTATG

GTTATGGAAG

GAGGCGTCCA

ACATTGCCAG

GCATTGGAGA

TCTGATGGCA

TTCGTTAAGG

TTGGATTCCC

GACAAAGTGA

GTCATAATTT

CTGCTTAAGA

GTAGCCAACG

TGTTACCGCA

TGGCGTCGAT

GCCAGAATGT

GTTCAAAAA-A

GGGTGGAGGT

ACAAGCGTAC

AATCCATTTA

AGGAGCTCTG

ATCAGGTGCA

TCGGAGTTTT

AGGTGGTAGC

CTGGAGTTGC

TTATTCAAAA

ACTACGTGAA

ACGACGACAC

ATGATTACAT

ACCCCAACTT

CTGCTTAGTT

TTCTGATATA

TTCGATCGTC

TGTTGATCCA

GCTGCCTGCG

CAAG TACATG

GGAACGAACA

GTTCCTGCGC

CACTGGCTAT

TGTGGATCCA

AGACAAAGTG

GGGGGAGAAG

CTATCATCCA

CCTCTGGTTT

GAGGACTTCT

GGAGGTCGCC

AGCTAGTCCT

*GGTGGTTTTT

ATGGTTTCTG

AAAAACTCGA

GTGAACGATA

ACGGACTTAA

GACTGGAAG T C TGGAAACAT

TGGAACCTGA

TTGATGGTAA

TCATTGCTCA

GGAGATGACC

GAGATCAAGA'

GATTGCTTCG

CGGCTGATGA

GGTCTGGTGG

CTGGAGGGTA

ACAGTCTTTA

CCGTCATTCT

CAATGTTTCT

TTAAAGCCAG

GACTAGTTCT

CGATGGGCTC

CTGACATCGA

TTGGAGTGAA

TCACCCGCCA

CACAGAGAGG

AGATGTCAGG

ATATCGCTCT

CCAAATTTGG

AAACTACAGG

ACTTTGCATT

AGTTAGTGCG

GAGGAAAGGA

TCAAGGCCAT

TGGGATATGG

GCTATG TAT C

ATCGTCATGT

AATAAAATTA

CTTC

INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 1075 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID TCGGAGCTCT CGAATCCTCA CAGGCCCTTT TTATTTCCCT GGTGAACGAT ACGATGGGCT 1, 1 WO 98/11205

CGCACGCTGA

120

AAGAATGGGA

180

AGCTTCAAAA

240

ATCGAGAGGA

300

GGGAGATGAT

360

GTGCTAAAAA

420

TCGCTCTTCC

480

GCTGGCCCTG

540

GGTTGGACTA

600

TCGTGGACGC

660

GCGTGGGGGG

720

ATCCCCACAA

780

TCAACTCCAT

840

GTGTCACTGT

900

GTATATGATA

960

TTTTCTGTTT

1020

AGCCGCTTC

1075

GAATGGCAAC

AACCAGGTTA

CGTTATGGAA

CGAGGCGTCC

GACATTGCCA

GGCATTGGAG

TTCTGATGGC

CTTCGTTAAG

TTTGGATTCC

AGACAAAGTG

CGTCATAATT

CCTGCTTAAG

GGTAGCCAAC

TTGTTACCGC

ATGGCGTCGA

AGCCAGAATG

AGTTCAAAAA

GGGGTGGAGG

TGACAAGCGT

GAATCCATTT

AAGGAGCTCT

GATCAGGTGC

ATCGGAGTTT

AAGGTGGTAG

GCTGGAGTTG

CTTATTCAAA.

AACTACGTGA

TACGACGACA

AATGATTACA

GACCCCAACT

ACTGCTTAGT

TTTCTGATAT

TTTCGATCGT

TTC-TTGAT-C

CGCTGCCTGC

ACAAGTACAT

GGGAACGAAC

AGTTCCTGCG

TCACTGGCTA

CTGTGGATCC

CAGACAAAGT

AGGGGGAGAA

ACTATCATCC

CCCTCTGGTT

TGAGGACTTC

TGGAGGTCGC

TAGCTAGTCC

AGGTGGTTTT

CATGGTTTCT

AAAAAACTCG

AACGGACTTA

GGACTGGAAG

GCTGGAAACA

ATGGAACCTG

CTTGATGGTA

TTCATTGCTC

AGGAGATGAC

GGAGATCAAG

GGATTGCTTC

AC GGCTGATG

TGGTCTGGTG

TCTGGAGGGT

CACAGTCTTT

TCCGTCATTC

TCAATGTTTC

GTTAAAGCCA

AGACTAGTTC

PCTINZ97/00112

ACTGACATCG

TTTGGAGTGA

TTCACCCGCC

AkCACAGAGAG

AAGATGTCAG

AATATCGCTC

CCCAAATTTG

AAkAACTACAG

GACTTTGCAT

AAGTTAGTGC

GGAGGAAAGG

ATCAAGGCCA

ATGGGATATG

TGCTATG TAT TAT CGTCATG

GAATAAAATT

TCTTC

INFORMATION FOR SEQ ID NO:56: SEQUENCE CHARACTERISTICS: LENGTH: 1962. base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

GTTTTCCGCC

AATCAATTGA

120

GGTCGAGCAT

180

TCATTCGTAT

240

GGCGACAGAC

300

CGGTCTGGCG

360

CATCGAATTT

420

CAATCCTTTC

480

TCATAGTTAC

540

TCGTCATCAC

600 SEQUENCE DESCRIPTION: SEQ ID NO:56: ATTTTTCGCC TGTTTCTGCG GAGAATTTGA TCAGGTTCGG *ATTGGGATTG

AAGGTTTTTA

CTGTACAGAT

TGCTTTGAGA

AGAACTTATT

AAGCTCGGGT

GCGTTTGTGT

TACAAGCCGG

CCTGGCAGCT

AATCGATGAT

TTTTCAGTAT

CGAAGCTTCC

GAGTAGCGGA

GCTTTTCAGA

TGCAGCAGGG

TCATGGGGGC

GCGAGATCGC

TATGTGGAGA

GCTCCCAAGG

TTCGATCGCC

CGATATCGAG

ATTCGCAGAC

GGTGGAACTG

GCAGGTTGTC

CTCTGTCCGG

CAAACAGGCC

AACTGGCCGA

AAGGTTGCCA

ATGGCCAACG

ATCTCCGACC

AGACCCTGTC

ATTTCTCGCA

ATGCTTCTCC

GGCGCCATTG

AAGGCCGCGG

TCTGCAGAGC

ACATATTTCC

GAATCAAGAA

ATCTGCCTCT

TGATCGATGG

AGGTCGCTGC

TTCCGAATTG

TGACCACGGC

GCGCGCGCGA

CACGATGTGC

GTTCTGACCG

WO 98/11205

AAGCCGACGA

660

CCTATTCTTC

720 TGTCCAGCGT1 780

ACGTGATACT

840

GCGCGCTCAG

900

TGGAGCTGAT

960

ACATCACAAA

1020

CCGGCGCTGC

1080

CCATTTTCGG

1140

CCTTCGCAAA

1200

CTCAAATAAA

1260

AAATCTGCAT

1320

CCGCTACAAT

1380

ACGAAGAAAT

1440

TGGCTCCTGC

1500

TCGTTCCTCA

1560

CGGAAATCAG

1620

AAATACACAG

1680

GAAAGGATTT

1740

ATTCCTTTGC

1800

GTGCAGAGTA

1860

CAACGCCCTA

1920

TTTTTTATAA

1961

AACCCAATGC

CGGAACCACG

TGCCCAGCAG

CTGTGTCTTG

AGCCGGGGCT

TCAGAAATAC

GAGCCCCATC

GCCTCTCGGG

GCAGGGCTAC

GAATCCT'ITC

GATCCTCGAT

CCGCGGACCC

CGATGAAGAA

CTTCATAGTC

TGAGCTGGAA

AAAGCACGAG

CGAGCAGGAA

AGTTTACTTT

GAGAAGCAGA

CGATAATTAT

AGCGCCCTAT

CACTCTTGCG

ATTTACTGCA

CCGGCCGTGA

GGGCTCCCCA

GTCGATGGTG

CCTCTTTTCC

GCGACCCTGA

AAGGTTACCG

GTTTCCCAGT

AAGGAACTCG

GGCATGACAG.

CCCGTCAAAT

ACAGAAACTG

GAAATAATGA

GGCTGGCTCC

GACAGAGTAA

GCTTTACTTG

GAGGCGGGCG

ATCAAGGAAT

GTGGATGCGA

CTGGCAGCAA

AGGATTCCTT

AAGGAGAGAG

ATCGCTTTCA

CTTCTCGTTC

CAATCCACCC

AGGGCGTGAT

AAAATCCCAA

ACATCTATTC

TTATGCAGAA

TTGCCCCAAT

ACGATGTCTC

AAGATGCCCT

AAGCAGGCCC

CTGGCTCCTG

GCGAGTCTCT

AAGGATATAT

ACACAGGCGA

AGGAGAT TAT

TTGCTCATCC

AGGTTCCGGT

TCGTGGCAAA

TTCCTAAGTC

AATGAAAATG

TCTGTTCACT

AGAGCTTATC

ATATGCATAT

GGACGATGTC

GTTAACGCAC

TCTGTATTTC

TCTCAATTCG

ATTCAACCTC

TGTGCCTCCA

GGCCGTCCGG

CAGAGAGCGT

GGTGCTGGCA

.CGGAACAGTC

CCCGCACAAT

TAACGACCCG

CGTCGGGTAC

CAAATATAAG

GTCAATCGCT

GGCGTTCGTG

GCAGGTGATT

GCCGTCCGGC

AATTTCCATA

TCTATTTATA

AATTGTATCA

TACTATAAAC

PCT/NZ97/00112

GTGGCGTTGC

AAAGGCCTGG

CATTCCGATG

GTTCTCCTCT

ACGACCTGTC

ATTGTCCTGG

ATAATCATGT

TTTCCCAAGG

ATGAACCTAG

GTCCGGAACG

CAAGCCGGCG

GAATCCACGG

ATTGACGATG

GGCTTCCAGG

GACGCAGCAG

GTGAAGTCGT

TTCTACAAGA

AAGATTCTGA

TGATTCTAAG

TAATAAAGTG

TATGGATTGT

GATATATGTT

INFORMATION FOR SEQ ID NO:57: Ci) SEQUENCE CHARACTERISTICS: LENGTH: 1010 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: GACAAACTTG GTCGTTTGTT TAGGTTTTGC TTGCAGCATT AAGCAAAGAA GATGAGTTCA 120 TTCCAGAGAA TATAAGTCTT TTCCAGTTTG 180 AGGTGGCCCT CGTGGAGGCC TCCACAGGGA 240 SEQ ID NO:57: TGCAGGTGAA CACTAATATG GAAGGCCAGA TTTTTCACAG CCCTTTTCCT GCAGTACCTG TTCTGGAAGG TGCTGAGAAA TACCGTGATA AGGAGTACAA CTATGGTCAG GTGATTTCGC WO 98/11205

TCACAAGGA.A

300

TTGTTCTGCT

360

GCGCAGTGTT

420

AGGATTCTGG

480

TGAAACTGCC

540

AAATTTTTGA

600

TGTGTGCACT

660

ACAGAAATCT

720

GAAATTTCAC

780

GTTGCGCCAC

840

ACTTTATCAG

900

TGCTCTCCCT

960

TCCAAAGCTG

1010

TGTTGCAGCT

TCCAAATATG

TTCTGGGGCA

AGCAAAGATT

TGTTATTATT

GAGAAACTAT

CCCTTATTCC

GATTGCAAAT

CACGTTGGGG

TCTTCGCAAC

TTCTTTGATT

CCGGTTTAAA

TTCATGACTG

GGGCTCGTGG

GCAGAATACC

AATCCTTCTG

GTTGTGACAG

GCAGATAACG

GAGGCCGCAG

TCTGGCACCA

CTGTGCTCTA

CTGATGCCAT

GGAGGCAAGG

ACTTATGAGG

AATCCTATCG

CGGCTGCTCC

ACAAAGGCAT

CCATTATTGT

CACACATCAA

TTGGGTCTGC

AGCATGTCAT

GGCCTTTTGT

CAGGGGCCTC

GCTTGTTTGA

TCTTTCACAT

TCGTGGTCAT

TCAACTTCGC

TTAACGAG TT

TCAAAAGGGC

GCTGGGAATA

TGAAGTTGAA

T TAATGAGAAG

GAACACAATT

ACAAATTTGT

TAAAGGTGTC

TGTCCATGAA.

ATATGGCATC

GTCCAGATTC

GCCTATTGTC

CGATCTCAGC

PCT/NZ97/OO1 12

GATGTTGTAT

ATGTTGGCCG

AAACATATCC

GTGAGGCAAG

CCATTGCAGG

CAGGATGATC

ATGCTCACTC

TCTCTTGTAG

ACGGGCATCT

GATCTCCGAC

CCGCCTATAA.

CGCTTGAAAC

ACTGGCGCCG GATCTACTGC INFORMATION FOR SEQ ID NO:58: SEQUENCE CHARACTERISTICS: LENGTH: 741 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:58: GAATTCGGCA CGAGACCATT

GTCAAAGGAG

120

TACCCAGATG

180

CGAGATTCTC

240

GCATTTGCCA

300 T TTAGTATAG 360

AACAAATACT

420

TCTTTGGA.AA

480

CCAAAATCAT

540

CAGTGAATAA

600

GTGTTAGTGA

660

AATCTTGATG

720

AAAAAAAAAA

741

ATCAAACAAA

TGAAATATAC

TTCCACATGC

AATTGTGGGT

TATGACGAGC

CACCTGTGGT

CCGCTTAGTG

GGCTGATGTG

TTTTGTTAGA

ACGGAATGAT

GATTGTGTCT

TCCAGCTAAT

TTTTGAA.ATT

CACTGTCGAT

TTCAGAGATA

TATAATCCTT

TAGGCACTGC

TTGTTTTCTT

TGGAATGCTA

AACTGGTTGT

GTGTTTAGAT

GTCAAATCTT

TTTTCAATGG

ATTGGCATAG CAATTGGTCA

GGACCTAATG

GAG TACCT CA CATAACAG TT

CGTAGGTGTT

AGATCCTTCA

TCTTTCTGGA

AGTACTAGTG

TCCAGAGGGT

CCATCTTTAC

GATGGGCTGA

TAAAAAAAAA

GTGTGGAGGC

GCAAATTTGT

TCAATCAATG

TGGCAGAACA

CACTTTTCTC

ACTTTGGTAT

TCCAGAGTTC

GTTTACAACC

AAGGCTATTG

CTGACTCTCT

AAAAAA

TTCTATCTTT

TAGTCAGCTA

GTGAAGTATG

TTTGTCCTAG

GAACCTCCTG

TTCCATAAGA

GGCAATAATG

TAAGGGAGTT

AACAGTTGTT

AGTAAGGTTG

TGTGATGTCA

INFORMATION FOR SEQ ID NO:59: WO 98/11205 WO 9811205PCT/NZ97/001 12 SEQUENCE CHARACTERISTICS: LENGTH: 643 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:59: CTCATCTCGG AGTTGCAGGC TGCAGCTTTT GGCCCAAAGC ATGATATCAG

GCAGATGAAG

120

ATGGCTTGTC

180

CCCAGCTGAT

240

GCTTGAGCTT

300

TTGCCACGGT

360 ACCAGCAGT1C 420 CGTTGTTTTT1 480

GATTGTCCAT

540

TGCTATGCAA

600

GAGCTTGCAG

643

CAAACGGATC

AAGATGCTCC

GATAGGTGGA

GTGAA.AGCTG

GTCTTTCACA

GAAGGGACGA

ACTTCTTCCA

GATGACTGCT

AAACCTTGGC

TGATAAATCC

AAACAGTTTG

TCATCAGAGG

AGTATGAGCA

ATATTCTCCA

TGGC"ITCAGT

GGAATGTGAT

TCGGCGCAGT

GGAGCGATTT

AGAGAAATCT

AGGCCTGGCC

CGTTACTGGA

TTACACTGTC

TCTGCGAGAG

TTACCAGAGC

TCTCAATGAT

GGAGGCCTGC

TTACATGAAT

GACTACTGCG

GCATGGGATA

TTAGGTCCCT

GCAGCGGGTT

AGAGCAGCAG

TTGGAAGGAG

T.TACTCACAG

GACCCTGAGC

GCAGAAACTG

CCTCATAGAG

TACAAACCAA

TTGCTAAGCG

TGA

ATCAAACGAC

TCATTGCCTC

TTCGGACCAA

CA AAAGAGAG

TCATCAGAGG

AAGTGATAGA

GGGTGAAGCG

ACCCGCTCGC

GAATTGGTAT

AAGGAATTTA

INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 441 base pairs TYPE: nucleic acid STRANDEDNESS: single (ID) TOPOLOGY: linear (X4)

GAATTCGGCA

ACGATGTCAG

120

TGGCATCTTG

180 G GGAT CC TG C 240

GACTGCAACT

300

GCTGCCATGG

360

TATGGTATGC

420

ACCATCTGGA

441 SEQUENCE DESCRIPTION: CGAGAATTTT TCTGTGGTAA CATAACAAAC TCCAAAGGAT GCTTATCAAG CGTCTCCTCC CAATGAGAAA AAGATGGCTC AATGAAAGCT GATTTAATGG TGTTTTTCAC ACAGCGTCTC TCTGGCCAAG ACTTTAGCAG CATGGTTGCA G SEQ ID NO: GCATATCTAT GGCTCAAACC TGGTATGCGT GACAGGAGCG AGTGTGGTTA CCAAGTGAGA ATTTATGGAA GTTAGATGGG ACGAGGGCAG CTTCGATGAG CAGTCGTGGG TGTCAAATCA AAAAAGCAGC ATGGGATTTT

AGAGAGAAGG

GCTGGTTACT

GGAACTGTGC

GCGAAAGAGA

GTCATCAGAG

GATCCCAAGA

GCCCAAGAAA

INFORMATION FOR SEQ, ID NO:61: Wi SEQUENCE CHARACTERISTICS: LENGTH: 913 base pairs TYPE: nucleic acid.

WO 98/11205 WO 9811205PCT/NZ97/001 12 STRANDEDNESS: single TOPOLOGY: linear (xi)

GAATTCGGCA

CAGGATCCTG

120

AATCCTCCGG

180

TGGCTCATCA

240 GGTAATCC7S 300

CTCTGGAA:-AG

360 GGTGTTT-'r- 420

ATTAAGCC;A

480

GTGAAGCGAG

540

ACACCAGGCA

600

AAAATGACAG

660

TTTGCAGAGG

720

TTCATTATGC

780

GAACCCCACT

840

TCACATATCT

900

GATGCTACCC

913 SEQUENCE DESCRIPTION: SEQ ID \.1:61: CGAGGAAAAC ATCATCCAGG CATTTTGGAA A7TTTAGCTCG

CAATGGCTTT

TCCATCGAGG

TGCGATTGCT

TAAAGACAAA.

CAGATTTGGA

ATGTTGCCAC

CAATCAACGG

TTGTTTTCAC

AAGTTTTTGA

GATGGATGTA

AGAACAAGAT

AGACCATGCC

ACATGATACT

TTGTATATGA

ATT

TGGCGAAGAG

AACAGTGTGC

TGAGCGAGGA

GCATCTGTTG

TGATGAAGGA

TCCCATGGAT

GGTCTTGAAT

GTCATCTGCT

CGAATCATGC

CTTTGTATCG

CGATCTCATT

ACCGAGCATG

GAGACAGGTA

ACATCCTGAA

CAGACTGCCT

GTTACAGGAG

TATAGTGTTA

GAT CTGC CGG AGCTTTGf.ATG

TT.CGAGTCCG

GTTATGAGAT

GGGACTGTGA

TGGACCAACC

AAGACATTAG

ACTCT TAT CC

ATCACAGCCT

CAGCTGCTTC

GCAAAGGGCA

TGCCACAAGA

CT'GCTGGGTT

GAGCAACTGT

GGGCAAATGA

CTGCCATTGA

AGGATCCCCA

CGTGTGCAAA

AT TTACAGA

TGGATCTTTG

CAGAGAAAGC

CCACATTGGT

TGGCACTGTT

ACTTGGATGA

GATACATCTC

CCCCTTGATT

AACGCCTTTG

CATACGGTCA

GCGAGACACT

GAGATTGACT

TGCTGTCAG

CAATGAC-ATA

AGCCAAGT-C

TGATTTCCAII.

CAGAAAA.L

TGCTTGGGAT

CGTTGGACCA

AACCCGGA.AT

TCTCTGTATC

TTCCACATGT

INFORMATION FOR SEQ ID NO:62: (iJ) SEQUENCE CHARACTERISTICS: LENGTH: 680 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (x i)

GAATTCGCCA

ATTGATCAGT

120

AGATGAAGCA

180

CGGCTCTTCG

240

GGACCCAGAAZ

300

GCTAA.ACCTZG

360

TTGTCAAGGG

420

GGAGCTTG:T

480 SEQUENCE DESCRIPTION: CCAGATCAAT TTTTGCATAT CACAGAGTCA TGGCCAGTTG TGCGAAGAGA ACAAGAGAGT CTGGTCATGA GATTACTGGA CACACAGGGA AGCTTGGGCA TTCAAGGCAG AGCTTAACGA GTTTTCCACG TTCCCAAGCC CGTCCTGCCC TGAGGGGAAC SEQ ID NO:62; TATTAAAAAG TAAGTGTATT TGGTTCCGAG AAAGTAAGAG GCTTTGTGTA ACTGGGGCAA ACATGGCTAT TATGTTCATC TTTGCTCG CTCCCAGCGC CGA.AATGGCC TTTGATGATC TGTTAATCTG GACTCAAACC AGTAAATCTG CTTCGACCCT CC TT CTC TAT

GGTTGAATGG

ATGGGTACAT

GAACTGT TAG

CAAGTGAGAA

CTGTGAGCGG

CTCTTCAGGG

GCGAACGCATC

WO 98/11205 WO 98/ 1205PCT/NZ97/001 12 GGGCACTGTG' AAACGAGTGA TACATACCTC 540 ACCTGACCCC CCTGATACTG TGCTGGATGA 600 AAAGACAAAG ATGGTCGGAT GGATGTACTA 660 CCATAAGTTC GGATCAGAGA 680 O TCCGTTTCA GCAGTGAGAT :7CACTAGGGALA ATCTCATTGG ACTTCGGTCG AGTATTGCAG CATCGCCAAC ACTTATGCAG AAGAGGGAGC INFORMATION FOR SEQ ID NO:63: SEQUENCE CHARACTERISTICS: LENGTH: 492 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: GAATTCGGCA CGAGGCTGGT TCAAGTGTCA

AGATTTCAGA

120

GAGCTGCTGG

180

TTAGAGGAAC

240

CTGGGGCGAA

300

ACGCCGCCAT

360

CCGAGGACCC

420

GATCGTGTGG

480

TGCTTTTTAC

492

AGAGCTGCTA

CTTCATAGGA

TGTGCGAGAC

TGAGAGGTTA

TGATGGTTGT

CGAGAACGAG

GAAAACCAAG

GG

AATCATGAGA

T CAT GGCT CG

ACTGGTAATC

ACTCTCTGGA

GAGGGAGTTT

ATAATTAAAC

TCTATGAAGC

SEQ ID NO:63: GCCCAAGGC 'C.TCCCCTACA TCCATCAAGG AAGTACAGTA TCATGCGTTT GCTTGAGCGA CGGTGAAGAC GAAGCATCTA AAGCAGATTT GGATGATGAA TCCATGTTGC CACTCCCATG CCGCTGTCAA TOGGATOTTG GAGTTGTTTT CACGTCGTCT

GAGAATCCCC

TGT GTGACAG

GGATATACTG

TTGGATCTGC

GGAAGCTTTG

GATTTTGAAT

A.ATGTTTTGA

GCTGGGACTC

INFORMATION FOR SEQ ID N0:64: SEQUENCE CHARACTERISTICS: LENGTH: 524 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

GAATTCGGCA

TCCAAGCTTT

120

GGCTTCATAG

180

ACAGTTCGCA

240

AACGAAAGAC

300

GTAGATGGTG

360

CGCTTGAAGG

420

TGTTCAAGAT

480 SEQUENCE DESCRIPTION: CGAGCTTGTT CAAAGTCACA TCGTCTACCT CCCTGAAAAG CTGCTTATCT CATTCGTAGT ACCCAGATAA -TOTGGAGAAG TCAACATCGT GAGAGCAGAT TAGATGGAGT ATTCCATACT AAACCCTAAT AGATCCTTGT CACCTTCAGT AAAGCGGGTG SEQ ID NO:64: TATCTTATTT TCTTTGTGAT ATGAGCGAGG TATGCGTGAC CTTCTCCAGA AAGGTTACAG TTTAGTTATC TGTGGGATCT TTGCTAGAGG AAGGCAGTTT GCATCACCTG TCTTAGTCCC GTGAAGGGCA CTATCAATGT GTGCTTACAT CCTCCTGCTC

ATCTGCAATT

AGGAGGCACA

AGTTCGCACT

GCCTGGTGCA

TGATGCAGCA

ATATAACGAG

CCTCAGGTCC

ATCAATACCG

WO 98/11205 WO 9811205PCTNZ97/OO1 12 ATACGACTAT AATAGCTTAG AGCGTTCCCT GCTGGACTGP% GTCA 524 INFORMATION FOR SEQ ID N0:6S: Ci) SEQUENCE CHARACTERISTICS: LENGTH: 417 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

TCCTAATTGT

GTTIGTTCATG

120

ATAACGAGAG

180 TCTG GAC C CC 240

AGGTTTGTGG

300

GAGAGACGCT

360

CTATGCAGAIT

417 SEQUENCE DESCRIPTION: TCGATCCTCC CTTTTAAAGC CAGTGCTAGC AGGAGGAGCA GACAGAAGTA AGTTTGTGGA TCTGAGGACA ATGGCAAGCT ATTGTTCAGG GCCTTCTTCA GGCGAGGTTG AGTCTCTCAG GTCTTGGATT ATCACAGCAT SEQ ID N0:65: CCTT'CCTGG CCTTCATTCC GCGTTrGCAAT TGGGGAAAAT AATAGCAA_'CC ATGCCGGTGT CGTTTGTGTC ATGGATGCGT ACGAGGCTAT TCAGTGCATG AAAATTGCAT GGGGATCGAT TACTGATGCG CTCAAGGGCT

AGGTCACAGA

T CCA A A 2,

TTCCTTCTIGG

CCAGTTATGT

CCACGGTGCA

TGCAGATCTT

GTTCTGG

INFORMATION FOR SEQ ID t4O:66: Ci) SEQUENCE CHARACTERISTICS: LENGTH: 511 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

ATGACACGAA

CTTCATTCCA

120

TGAAGAAAAT

180

GAGTTTCCTG

240

CGCAATTCCG

300

GGGGAAGCTG

360

CAATGGTTGC

420

GGAGTATCCG

480

GAATTTAAGG

511 SEQUENCE DESCRIPTION: TTTGTGCCTC TCTCTGACCA TCATCCAGGA GCTTCTGTTA GGATACGGCG CTTCCAATTC GGGATTCATA TCGCAAGAAT GTAACGCCAG AAGAGGCAGG GAGATATGCO AAGCCGATCT TCCGGAGTCT TCCACGTCCC GTATGATTAG TTTAATAGAT TTTTCTTAGA ATTTGGATAC SEQ ID NO:66: GAGCTTG-AAG CTCTCGTCTTC .TATCCAT TTC CTCAAAATGG TCGGAAATTA ATGTGCCTTA GCTGCTCGGC CGGGGTTACT CTCACTTATG GAATCCGAAG CTTGGATTAT CGCAGCGTTT TGCGCCCTGT GATCATCTGG TGACGGGGTA TCCTGTATGA

T

TCTGATATCG

ATGCCTACCT

CCGGGGGCTG

CAGTCCGTTT

AAGCATTATC

TCGGCAACAT

ATGGATTACA

ATTAGTTTAT

INFORMATION FOR SEQ ID NO:67: SEQUENCE CHARACTERISTICS: CA) LENGTH: 609 base pairs TYPE: nucleic acid STRANDEDNESS: single D) TOPOLOGY: linear WO 98/11205 WO 98/ 1205PCTNZ97OO1 12 SEQUENCE DESCRIPTION: SEQ ID NO:67: CATTGATAG: TGATGGAAGA CCATCAGTAA AGCATGAAAA AGAAATTGTT

GAAGTCAGTT

120

AATATGTAAT'

180 C TGAC C TT Ca 240

AGTGGCGAAC

300

GAGGAACAGG

360 CATTCCTTc: 420

CC'TTCAAGGC

480

TGGAGGCAAPT

540

TTCAAACAGG

600

TTTGGCCAA

609

GCTCCAGCAG

CCATAAACTT

CATATTTATT

CAACTT GACA

CTACATTGGT

TGTCAGAGAG

CTCAGGT GCT

CAAGAAAGTT

AT AT TTAT CC

AACCTTTTTA

ATGCAGGAAG

CCAATTCTAA

GGGTTGGACA

C-GTCATATAA

AC CT CCCCT T

ATTATACTCC

GATGTAGTTA

AGGO TAT TTA

GCAATTGTTT

TGCCTCGTGC

TATCTCTACT

TGC-CCAACAG

CCAAAGCCAG

CTAATCCTGA

ATGGATCTTT

TCTCGGCTGT

AA.GGGAGGGT

T TGTATCCTT

CGAATTCGGC

CGCTGTCTAC

CAGCAAGATT

CCTTGCTCTT1 GA AGGCTAAG

GGAGGACCAT

CAAGGGACCA

T GGAACCCAT

CCAAGGTGJX

TTTGCCTTTG

ACGAGAATCA

CTGATTTTTC

CTGATTATTG

GGTCATCCCA

CTTCTGGAAT

GCAAGTCTTG

CAGCTGACGG

CAAGAAGGGT

INFORMATION FOR SEQ ID NO:68: Ci) SEQUENCE CHARACTERISTICS: ()LENGTH: 474 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: GCAAGATAGG TTTTATTCTT CTGGAGTTGG

GCATAGCA.A:

120

GCTCTATGTA

180

TGCAGCAAAT'-

240

GGTGATGGGA'

300

TCATCCTACC

360

CGTCCAACAA:

420 C CTC TTAG TA 474

TAAGCAGTTG

GCTGCAGACA

TGTGAGATGG

GCCACAGGTT

TATGCTCTTA

TTGAAGGATG

AATACATTGA

CAGCCATGGC

TGGTGGAAAA

AGAAGCCTCT

ACATTGGCCG

TACGCCCGTT

CCGGGGTCCA

AGGACATGGG

SEQ ID NO:68: GTGAGGCTTG GAAATTTAAG GGTCTGTGGA ACTGAAGTAG CAACACGTCT ATTGTGACCA TCTAAATTCC TCTGCCACCT TTTTGTTGCC CAAGAAGCTG TGCTGCTTGT GACCTGGCCA TATCCTTTAT GGGTCTTTGA CCGTTGTTAT CTCTACCATT

TAAAAAGGGT

CTCATACTGT

CCTCTATGGC

CAAGAATAC T

TTGCTGCTGG

AAGCACAGCG

GTGATCACAA

GGAG

INFORMATION FOR SEQ ID NO:69: Ci) SEQUENCE CHARACTERISTICS: CAt) LENGTH: 474 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear SEQUENCE DESCRIPTION: SEQ ID NO:69: WO 98/11205 WO 9811205PCT/NZ97/00112

GCAAGATAGG

GCATAGCAAT

120

GCTCTATGTA

180

TGCAGCAAAT

240

GGTGATGGGA

300

TCATCCTACC

360

CGTCCAACAA

420 C CT CTTAG TA 474

TTTTATTCTT

TAAGCAGTTG

GCTGCAGACA

TGTGAGATGG

GCCACAGGTT

TATGCTCTTA

TTGAAGGATG

PLATACATTGA

CTGGAGT:T3G

CAGCCAT'-GC

TGGTGGA.AAA

AGAAGCC:'CT

ACATTGGCCG

TA'CO CC C CTT

CCGGGTCCA

AGGACAT!C CO C TG'GGCTTG

GGTCTOTGGA

CAACACGTCT

TCTAAATTCC

TTTT:GTTGCC

TGCTGCTTGT

TATCC'TTTAT

CC G1T GT T AT

G-.AATTTAAG

ACTGAAGTAG

ATTGTGACCA

T C TO CAC CT

CAAGAAGCTG

GtA-CCTGGCCA

GGGTCTTTGA

CT CTACCATT

T.;AAAAGGGT

CTCATACTGT

CCTCTATGCC

CAAGAATACT

TTGCTGCTGG

AAGCACAGCG

GTGATCACAA

GGAG

INFORMATION FOR SEQ ID N0:70: SEQUENCE CHARACTERISTICS: LENGTH: 608 base cairs TYPE: nucleic acic STRANDEDNESS: sinale TOPOLOGY: linear (x i)

CATTGATAGT

GAAGTCAGTT

120

AATATGTAAT

180

CTGACCTTCA

240

AGTGGCGAAC

300

GAGGAACAGG

360

CATTCCTTCT

420

CCTTCAAGGC

480

TGGAGGCAAT

540

ATCAAACAGG

600

TTGGCCAA

608 SEQUENCE DESCRIPTION: SEQ ID TGATGGAAGA CCATCAGTAA

GCTCCAGCAG

CCATAAACTT

AATATTTATT

CAACTTGACA

CTACATTGGT

TGTCAGAGAG

CTCAGGTGCT

CAAGAAAGTT

ATATTTATCC

AACCTTTTTA

ATGCAGGAAG

CCAATTCTAA

GGGTTGGACA

CGTC AT A -AA

ACCTCCGCTT

ATTATACTO-C

GATGTAGTTA

AGGGTATTTA

AGCATGAAAA

GCAATTGTTT

TGCCTCGTGC

TATCTC TACT

TGGCCAACAG

CCAA'AGCCAG

CTA.ATCCTGA

ATGGATCTTT

TCTCGGCTGT

AAGGGAGGTT

AGAAATTGTT

TTGTATCCTT

CGAATTCGGC

CGCTCTCTAC

CAGCAAGATT

CCTTGCTCTT

GAAGGCTAAG

GGAGGACCAT

CAAGGGACCA

GGAACCCATC

CCAAGGTGAA

TTTGCCTTTG

ACGAGAATCA

CTGATTTTTC

CTGATTATTG

GGTCATCCCA

CTTCTGGAAT

GCAAGTCTTG

CAGCTGACGG

AAGAAGGGTT

INFORMATION FOR SEQ ID NO:71: SEQUENCE CHARACTERISTICS: LENGTH: 1474 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:71: GAATTCGGCA CGAGAAAACG TCCATAGCTT CCTTOQCCAAC TGCAAOGCAAT ACAGTACAAG AGCCAGACGA TCGAATCCTG TGAAGTGGTT CTGAAGTGAT GCGAACCTTC GAATCTGAAA 120 WO 98/11205 WO 9811205PCT/NZ97/001 12

AAACTGTTAC

180

ATCTCAGAAA

240

ACTCTGATTT

300

GGCATGAA-

360

GAGAGCATGT

420

AGAGCATGGA

480

GCACACCTAC

540 GAAtCCCGGA 600 TTTTCAGC1C 660

GTTTAGGAGG

720 CGGT TAT CAG 780 CTTATCTTG7T 840

TAATGGACAC

900

ATGGAAAGCT

960

TAATACTTGG

1020

AAACTCTAGA'

1080

ACTACATCAA

1140

TGGATGTTGC

1200

TGCATGCAAG

1260

ATTTAGGAAC

1320

TTCAGATGTT

1380

TCCAATGTCT

1440

AAAAAAAAAA

1474

AGGATATGCA

GAAAGGACCT

AGTTCAAATG

GGTGGGGATT

AGGGGTTGGT

ACAATACTGC

TCAGGGCGGA

GAATC".TCCT

AATGAAG CAT

CGTGGGGCAC

TTCGTCTGAT

TAGCAAGGAT

CATTCCAGTT

AGTGATGCTG

GAGAAGGAGC

TTTCTGTGCA

CACGGCCATG

TAGAAGCAAG

ATGAATAGAT

TCGATACTGG

TTTTTAACTT

TCTGCCAAAT

GCTCGGGACT

GAGGATGTAA

CGTAATGAAA

GTAACAGAGA

TGCATTGTTG

AGCAAGAGGA

TTTGCAAGCA

CTGGAACAAG

TTCGCCATGA

ATGGGTGTCA

AAAAAGAAAG

ACTGAJAAAGA

GCTCATCCTC

GGCGTTGTTC

ATAGCTGGAA

GAGAAGAAGG

GAAAGGTTGGQ

TTGGATAATT

CTGGACTAGT

TTTTTGTTAC

GTATATGTAA

TAATATATGT

CCAGTGGCCA

TTGTAAAGGT

TGGACATC-TC

TTGGCAC-CGA

GGTCCTGTCG

TTTGGACCTA

GTATGGTGGT

C GGCC C CTC T

CAGAGCCC-GG

AGATTGCCAA

AAGAAGCCAT

T GATGGAAGC

TGGAACCATA

CAGAGCCGTT

GTTT CAT TGG

TATCATCGAT

AGAAGAACGA

AGTCTGCAAT

AGCTTAACAT

TTTAGTTTAG

AGATCAATTT

ATTCGTATTT

AAAA

CTTGTCCCCT TACACTTACA CATTTACTGC GGAATCTGCC TCATTACCCA ATGGTCCCTG GGTGAAGAAA TTCAAAGTGG CAGTTGCGGT AATTGCAATC CAATGATGTG AACCATGACG TGATCAGATG TTTGTGGTTC GTTATGTGCA GGGGTTACAG GA'-AGAAATGT GGGATTTTGG AGCCTTrTGGA'CTCCACGTGA GGAAGTCCTC GGCGCCGATG AGCAGAGAGC CTAGATTACA TCTTGCCCTT CTGAAGACAA GCACTTCGTG ACTCCTCTCT CAGCATGGAG GAAACACAGG GATTGAGGTT GTGGGCCTGG TGTCCGTTAC AGATTTGTGG CAATCAATCA GATCAATGCC GAAAGGGAAA TTAAATTTTT CTTTTGTGAG GTTGAAACAA CTCGTG-ACAG TAAATAATA-A TTATATGAAA. AAAAAAA INFORMATION FOR SEQ ID NO:72: Wi SEQUENCE CHARACTERISTICS: LENGTH: 1038 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: GAATTCGGCA CGAGAGAGGG TTATATATCT TGCCAAGCTC TGGGCCACGG ATTTGGAATC 120 GGCGAATTTC ACAAAGTATT TCACCGATAA 180 CTTTGAGGGA AAAAAACCCT GCTACTTCAA 240 SEQ ID NO:72: TGATTCTGAC CTGATTGTCG TCGACGACAT TCGTGTCCTC GGGGCACCAG AGTACTGCAA TTTCTGGTGG GATCCCGCAT TATCCAAGAC CACAGGCGTA ATGGTGATCG ATCTTGAAAA WO 98/11205 ATGGCGGGCA GGGGAATTCA 300 CCGTATCTAT GAGCTCGGAT 360 GCAAGTCGAT CATCGTTGGA 420 CCGAGATCTT CACCCTGGAC 480 GCTACGCCTG GAATGCCAAG 540 TTTATCGATC AACGTATTAC 600 ATCGAATTAA ACCTGATTTG 660 GTTTTGAATT TCAATTCTGG 720 CAAATCCATC ATGAGGGACC 780 CGCCTGTGAA GAATGATATT 840 CAGCCAGCAG AGAGGCAAGC 900 AATTTTCGGC GACTGTACAG 960 CCTGAACCAA CAACTGTATA 1020 AAAA~AAA AAAAAhAAA 1038

CAAGAAAGAT

CAT TACC GCC

ATCAGCACGG

C-TGTCAGTTT

CGGACTTGCC

CTAAATGGGT

ATAAAATGCC

TAACGAATAG

AATCGTTTGA

GTGGACTGAT

AATGCCGCTG

CGATGTAAA'TT

ATACCTTATA

CGAAATC-IGG

ATTTTTACTG

TTTAGGCGGA

GTT GC AT T GG

CTCTGGATAC

GAGAGAGCCT

AAATAGAACT

AAGAAAACAA

ATTTAGTATT

CTATTTATAT

CAAGTCATGT

T TTGGAACAT

AATGTATCTG

AJGGACATAC

GTA'TTT.GCTG

GATAATTTGC

AGTGGT3AAGG

TTTATGGGCT

CTCTCCTCGG

T TACGCC TAT

TAGCACAGCC

AATAAGGTTG

TTGTACTGCC-

AGGGAAGGCG

TAATATCATT

CAACTCCATT

PCTNZ97/0O1 12

.!GAAGGAACG

rGTTTGGTTAA

AAGGCCTTTG

GCAAACCTTG

C-CTTATGATC

GGJTGCTTTTT

GCATCTTTCA

ACAGGCAGGJA

-:TCCATATAA

AZTGCCATCCT.

TTGTGAACT C .a.TGATAAGTT

'TGCATAAA

INFORMATION FOR SEQ ID NO:'73: SEQUENCE CHARACTERISTICS: LENGTH: 372 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) CTAGGGG TCT

CATGCAAGAG

120 GTCCTTT TAG 180

CGGGAGGAGG

240

CCATCGGAAT

300

TGGGCAATTC

360

AAATCTGCCA

372 SEQUENCE DESCRIPTION: TGGGGGGTTC CTGATGCCCA ATCTGTAGTC AGTAGTCTTG GGTAACATCA TTCCAACCAT AGCAAGATAT TCAGCATTGC TCAGCCGAGC TCGCCCCCTC TGGCTCGAAA TCGCCAAATT

GT

SEQ ID NO:73: ATTGTTGCTG TGCTTGG-AT TTGGATCTAT AGCTTT-TAGA ATCCAGTTCC ACCACCGGCT TTTGGGCACC AGATGGATAG AGTCCAATCG TCGTGAAAAT ATGGGCTACA ACAGGATTAA GAACCCAkAA

AAAGAGTCAC

ACACCTTCAA

GCATTATTTT

CCCTCAAAAT

AATTGCACAG

INFORMATION FOR SEQ ID NO:74: SEQUENCE CHARACTERISTICS: LENGTH: 545 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:74: WO 98/11205 WO 9811205PCT/NZ97/0O1 12 AAAGAATTC'G GCACGAGGGC AATCCGAGCC TAGCCA.ACCA ACTTGGCAGC AGGAGCACA

GGGAGTTGGC

120 C CAACAAGC C 180

ATCCTGATAA

240

GAAACATAAC

300

CTGAAGTGGT

360

ATGCCATTGT

420

TGACCATTCC

480

TTGTAATCTT

540

GTTTT

545

GAGAGAAGCT

TTTGCTCCCT

TCTGGGT TAT

CGTAGGAACT

TTATGAGCAA

GGTTGTGGGT

CCTAGGCGGA

GATATCTGGA

GTTAGGAAAT

TTGGAGAAGA

CAGTGTGGTG

ACAATTCTGG

AATCCAGATG

GAGGCACCAT

GGGGACACGA

AGGCCACTTG

C 'TTTGGTA T 1

ATGCTTCICAA

GATGGACGAT

AAGCTATCAA

CTAACTATGT

ACGCAGAAAC

TTAGACGGT

TTATTGAACC

'QTTGAAAAAT

GGTTCTTGTT

GGAATGGCAA

ACTAGCTGTC

CAAAGGACAA

GTTTGGAGAC

C TGTG CC TCC

TTATCTTCCA

'GAAGTCAG

GCAGGAA CCC 2GATTAAGTG A %GCCCCTCTA

CGGTTTT'OAT

A-ATCTTAATT

:TGAAA.TrCC

:TCGGTGGATC

INFORMATION FOR SEQ ID N0:75: SEQUENCE CHARACTERISTICS: LENGTH: 463 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

GCAGGTCGAC

CTAGTGATGA

120

TGTAGAGCCT

180

CGGTGGATTT

240

TCGTGCTTTT

300

ATATTTTTCT

360

CTGCACTGCG

420

ACATAGTACC

463 SEQUENCE DESCRIPTION: SEQ ID ACTAGTGGAT CCAAAGAATT CGGCACGAGA AAAAACAAAT GCTTTACGTA TACCTGGCCT TTTATACATG GATCTGAGTT TTTGTTACTC TGTATCACTG GGACTTGCCA CAAGCTCTGG CGTAGCAAAA AAGTTGTGGA TGACTTT'UGC ATATTCTCAG GGAGACCGTCu TGAAGTACTG GGTAACTGTT AACGAACCGT TACGATGTGG GGCTTCACGC ACCGGGCCGC TGTTCGCCTG GGAAATTCAG CGACAGAGCC TTATATTGTA GCCCATAACA GCTGTTAAAA ATATATAGCA*TAAATACCCA

GGG

CT TAGCTAGC

TTTATGCAGG

AGGACGAATA

t-AGAATGCTT

TGATCTTCTC

GATTTGGAAA

TGCTTCTTGC

INFORMATION FOR SEQ ID NO:76: Ci) SEQUENCE CHARACTERISTICS: LENGTH: 435 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:76: ACACTAGTGG ATCCAAAGAA TTCGGCACGA GGCTACCATC TTCCCTCATA ATATTGGGCT TGGAGCTACC AGGGATCCTG ATCTGGCTAG AAGAATAGGG GCTGCTACGG CTITTGGAAGT 120 TCGAGCTACT GGCATTCAAT ACACATTTGC TCCATGTGTT GCTGTTTGCA CAGATCCTCG 180 WO 98/11205 WO 98/ 1205PCT/NZ97/001 12 ATGGGGCCGC TGCTATGA-A GCTACAGTGA 240 GATTATCGTT- GGCCTGCAAG GGA'-ATCCTCC 300 AGCTGGACAG TCAAATGTT- CAGCTTGTGC 360 CAAAGGTATC GATGAGAATA ATACTGTTAT 420 ATTACCCCCA ATTTT 435

GCA'CC~'AATTOTCAAGG

TGCTAATTCT ACA.AAAGGGG TAAGCAT'TT GT GGGTTATG CAACTATC:AA GGGTTATTTC C AT G AC TA rC:GCCTTTTAT! 2TGGAACAAc AACAT TCCAA INFORMATION FOR SEQ ID ,'JO:77: Wi SEQUENCE CHARACTERISTICS: LENGTH: 4511 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (x-4

GAATTCGGCA

TACAAAGG;A

120

CACTGCTTA:

180

TGGACTGG'A

240

ACTCCAATTS

300

AACACTATGG

360

GACACTTCCA

420

AAATTTGATT

451 SEQUENCE DESCRIPTION: CGAGCCTAGA ATTCTATGGT CAGTCCCAAA, TGGTTAAAGG TACATGTATG ATCCTAAACA TACAGGCTT.T GCATATGCTC GCTTTACATT GTGCCTTGGG AAATCCAACT ATGATTCTCT GCAGGACTGC ATGATACCAT AATGCACGTG AATGACCGGG SEQ ID \10O: 7: GAAAATC'TT GGGACAAGGC TTCA.ATAC-AC TATCTAGGCGu ACCTAAACAA AATGTAACAG GCAATGGAGT GCCTATTGGA GTCTATACAA GGCCGTCACA CTGAADAATGG AATGGACGAC CAGGGGTAAC TACTATAAAA 7GCCCAA.G:T

:TAACCA.ATA

-ATTACCAGAC

CCAAGGGCGA

T ACGTAAzA-AG

C-TGGAAACGT

GCTATTTGCA

2) INFORMATION FOR SEQ ID NO:78: SEQUENCE CHARACTERISTICS: LENGTH: 374 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear

CTGCTCTGCA

CTTGAGGAAA

120

CTTCAGAAAA:

180

GCTCCGCCGC

240

AGCTCTTAG-C

300

AGCTGCTA!CA

360

GACAGGAC;A

374 SEQUENCE DESCRIPTION: AGCAGTACTA TGCACAGCAA CGCTCAAGCA TTGCTGAGGC ATGGCAATGG CACAAGCATT AACATTCTGC CGGAGGATAA CTGCTCTCAT CAAAAGCCTT AATTCAACAA TTGTGTTGCA

TCTG

SEQ ID NO:7

GGCCTGCTTA

CACCGTTTAT

CAGAGGCCGT

AAGCTTTGGA

CATCTCTTTC

ATCTCGAAAC

ACTGAAAACA

CTAAATAGCG

GTCTTGCAAG

TCCGCTGCTT

TCTGTTGAAC

TTTTCTGCA.A

GAGCGCTGAG

CAACATAGGG

CTGCCCGTTT

C-TCCTAGACG

CGGCAT CC CCT A AGGTAAAAA INFORMATION FOR SEQ ID NO:79: WO 98/11205 PCTNZ97/00112 SEQUENCE CHARACTERISTICS: LENGTH: 457 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

GAAGAATGGA

TCATGTTTCG

120

AAGGCCCAAG

180

ACGGTGATGT

240

ATATGGGCGT

300

AAGGAGAGAT

360

AGAATGGA.T

420

GATGAATA-

457 SEQUENCE DESCRIPTION: AGAGATTAAT GGTGATAACG GATAGCAACT TCTGCTTATC CATCTGGGAC TCATTTTCAC AGCAGTGGAT CAGTATCATC GGCTACCTAC AGATTCTCGA CAATGAGGAA GGAGTAGCCT* CCAAGCGTCT GTCAACTTTG GCGGATTTCT GAGGCCAACC SEQ ID NO:79: CAGTAAGGAG GAGCTGCTTT AGTGTGAAGG AGCTGCCAAC GAACACCAGG CAAAATTCTT GTTATAAGGC AGATGTAAA.A TTTCATGGCC TCGTATATTT ATTACAATAA CCTCATCAAT TTTCACTGGG ATACTCCCCA

ATTGTGA

CCTCCAGGTT

GAAGGTGGAA

GATGGAAGCA

CTGATGAAAG

CCAAAGGGAA

GAACTCCTCC

GTCTCTGGAG

INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 346 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (x4)

GGTGTGATGG

ATGCTGCACA

120

GGTTTCAAAT

180

GGCAAAGGCC

240

AATATTGGGA

300

AATACATGGG

346 SEQUENCE DESCRIPTION: CAGGAATTCC AGTCCTAAGG TTGTAGCTGC AGTAGCTTCA TTGGTGCAGG GTCATCTGCT CAAGCATTTG GGATACATTC TGTTGCAGTA GATCAATACC AATGGACGTC TATCGTTTCT SEQ ID CCATTTTGCA TCTGTTTGCT TTCAGTCTAC CCAAGGCTAG GTAGAAGCAG CTTCCCAAGG TATCAGGCGG AAGGAGCTGC TCATGAGGGT TCCCACACTC CAGGTAAAAT CGCTGATGGG ACCGTTATAA GGAAGATGTG CAGCTTCTCA CTATCTCCTG GTCACG INFORMATION FOR SEQ ID NO:81: SEQUENCE CHARACTERISTICS: LENGTH: 957 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:81: GAATTCGGCA CGAGAAAGCC CTAGAA.TTTT TTCAGCATGC TATCACAGCC CCAGCGACAA.

CTTTAAC:C AATAACTGTG GAAGCGTACA AAAAGTTTGT CCTAGTTTCT CTCATTCAGA 120 WO 98/11205

CTGGTCAGG:

180

CTTGCACTCA

240 TGGAAGCTrS 300

TCAAGCAAGT

360

TGACCCTCTC

420

AACTCCATGT

480

ATGGGATGGT

540

ATATAGATAC

600

AGCAGATTTC-

660

ACATAGATGIA

720

TCATCTTCAA

780

TAGTACTGTG

840 A.AAAT CTC;A 900

TGACATTTGA

957

TCCAGCATTT

GCCCTACATT

TGTCAACACG

TTTGTCATCT

TCTTCAAGAC

TCTGCAGATG

GAGCT"TCAAT

TGCAATTCGG

GTGTGAT CAT

TTTTGATACT

GACTCGCTTA

GCT GAG TCCA

ATTTCTCGAT

GCAC CT CGAG

CCA-PAATACA

GATTTAGCAA

AACACAGAGA

CTTTATAAAC

ATAGCAAGTA

ATTCAAGATG

GAGGATCCTG

AGAATCATGG

TCCTACCTGA

GTTCCCCAGA

TATTCATTAC

GAAAGGATCT

GTCTAGTCTT

TGAACTACAA

CACCTGCTG T

ACAACTACAG

AGTTCAAGAA

GGAATATTCA

CGGTACAGTT

GTGAGATTTT

AACAGTACAA

CACTATCAAAk

GTAAGGTGGG

AGTTCACAAA

TTTCTATIGTG

CTCGGTATTA

GATTTTGATT

AGTTGCATGT

TGTjCCAAAGA

TAGTGGGAAA

TGATAGTAAT

GAGATTGACA

GGAGACTGCT

TGCAACCATA

AACATGTCAG

GAAGCTCACC

GAGAGAGCGT

TATGTAACAA

AATTGATAGT

TCACTTGACA

AT GAATGCGA

TA-AAAAAAAA

PCTINZ97/001 12

AATTTGAA.AT

ATTTCTGTAT

TTGGGGTTAG

CAGACATATC

AAGCAGGCTG

AATCAGAAAG

ATGACTGA.AT

ACAGTAGATG

TCAAGATTTG

ATGATGTAA

CTGTTAACAA

TGCCATCAA-A

CTTTTAGTTG

INFORMATION FOR SEQ ID NO:82: SEQUENCE CHARACTERISTICS: LENGTH: 489 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (x

GCAGGTCGAC

ATCCTCCA:-T

120

GGACTAGAGT

180

GCATCTTCAG

240

GCCAAGGTGT

300

CTGTCGAGIAG

360

CAGACAATCC

420

TGAAAATGGC

480

TACCAGTAA'

489 SEQUENCE DESCRIPTION: SEQ ID NO:82: ACTAGTGGAT CCAAAGAATT CGGCACGAGA TAAGACTAAT TTTCCAGACA

CCCATTCAAT

AAAAGT CCTT

CACAGACAGC

TGGAAACTAC

AAACACTGTG

AGGGGTTTGG

GTGGGTAGTA

TACACTGGTA

CCCTTTAACA

CACCCTGTCC

AATGAATCAA

GGAGTTCCCA

TTCATGCACT

AAGAACGGAA

CTCCACCCAA

CAACTGTTCA

ATCTCCATGG

CAGATGCACC

AAGGAGGTTG

GTCATTTGGA

AAGGGCCCAT

TA-ATACACAG

ATTGATTCTT

TTTCAATTTC

AAATTTTAAC

GGCTGCTATA

GGTTCACACA

CGATTTTCCA

GCTGTGAATG

CAAGACACCA

TTTGTGGTGG

CTCATTGACC

AGATTTCGTG

TCGTGGGGAC

CCCGGGTGGG

INFORMATION FOR SEQ ID NO:83: (i4 SEQUENCE CHARACTERISTICS: A)LENGTH: 471 base pairs TYPE: nucleic acid STRANDEDNESS: single D) TOPOLOGY: linear WO 98/11205 WO 9811205PCT/NZ97/001 12 (xi)

GAATTCGGCA

ACAGACATAC

120

TATTCCAACG

180

GAGGGAAGCT

240

AACAGTGCTA

300

TTCGTTCCTC-

360

TGTCCGGGGC

420

ATCATTTGTC

471 SEQUENCE DESCRI PTION: CGAGAAAACC T-T.TCAGACG TTCTCACTGC CAATCAGGCT GGCAAGGAGT TCCCTTCGAT CTAAGACTTC AACTCCAGTC CTAGCTTCGC TAATGGTCTT AGAGTGTGGA GGAGAATCTG AGTCTTGTGG AGGTCCAACG CCGCAACCAC TTCTTCCAAT SEQ ID NO:83: AATGTTCTGA TGCTCGGCCC ACAGGTAGAT ACTACATGGC AACACCACTA CCACTGCCAT ATGCCTAATC TTCCATTCTA AGAAGCTTGG GCTCACACGA TTCTACACCA TCGGTITTGGG GATCAAGATT TGCAGCAAGT CCTTCAAGCT CAGCATTTTG

CGGCCAGACA

TGCTCGAGCA

TTTAGAATAC

T. ACGACACC

CCACCCAGTC

GTTGATCAAA

ATGAATACAT

G

INFORMATION FOR SEQ ID NO:84: SEQUENCE CHARACTERISTICS: LENGTH: 338 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (x i)

GTTCGGCACT

ATCTCTTTCA

120

AATCAGTGCG

180

TTGCTGTAAC

240

CAGGCTATGC

300

CATGTACGTG

338 SEQUENCE DESCRI PT ION: GAGAGATCCA TTTCTTTCAA GGAATATATC GTGCTTGCAG TCTATCTTCT GCTCTCCTTG AAACGCAGAT GTCCACAATT AATAAGCGTA TAATCGCCAC ATGGAGACGT TGTTAATTAT SEQ ID NO:84: TGTTGAGACA GTGAGTAGTA TTAGTTTGAT GATCTTTAGT TTCTGCAACA ATGTCGTTGC TTTTGCTACT AGCATTTGTT GCTTACTTAG ATACCTTCAT TATTAGAAAG AGACAGTTAC CGTCAATGGC AGCTACCAGG CCCAACTATT

CAAAGCTT

INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 1229 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi)

AGAGAAATA.A!

TGCAGGAGA

120

GTCTTTTCTC-

180

CAAAGGAATT

240

TAAGCGAGGA

300 SEQUENCE DESCRIPTION: TTATATTTGT AA4ATTTAAGT AAAACAAGCA TGCTGTCTAC GTGCCGAATT CGGCACGAGA TTGTGGGTCA TTTGCAGGTG GTACAAGGCT GCCATTGACA SEQ ID CTACGTTTAT TAAAAAACTA CAACCCTAAA TGAAGCTTAC AAATCAAATC CCTGCGATAT AGATCTTGGT TCGAGTCTCT CAGCTCTCTC AAGACACCAT GGTGAAGGCT TATCCCACCG AATGCAAGAG GAAGCTCCGA GCTCTCATTG *1.

WO 98/11205 PCT/NZ97/00112

CAGAGAAGAA

360

ATGTCAAGAC

420 .ACG TC T AA 480

TCCCCATAAT

540

CCGGGGGACC

600 AAGC CGCCC T 660

TGGGGTTGAA

720

ACAAGGAGAG

780

CTTACTTCAC

840

AGGCACTGCT

900

ACGCTTTCTT

960

ATGCGTAGAT

1020

TCATCTAATC

1080

CTTGAACCTA

1140

ATTTTGGAAT

1200

TAAAAAAAAA

1229

CTGTGCGCCG

CAAGACCGGA

CAGTGGTCTG

CACC TAT GCT

TGACATTCCG

TCCTGATGCT

TGATAAGGAA

ATCTGGTTTT

AGAGCTT-QTG

TG CT AT CCT

TGCTGACTAT

TCATACCTTC

TTITTCGATTA

CATGTTTTTG

CTGGTTGTGT

AATAAAATAA

ATCATGCT7-TC GGGCC CT T C C GACATCGCAr'G

GACCTTTATC

TTCCATCCTG

ACAAAAGGAC

.:,TTGTGGCCT

GAAGGACCAT

ACTGGAGAGA

AGTTTTGCAG

GCGCGAAG CT C

TGCAGAGACA

TATAGTCACA

AAAAGTTCG

TCTATCA.AGC

GAATCGC~aTG

GGACGATGAG

TTAGGTC.-

AGTTGGCTGG

GAAGAGAAGA

CT CAT CATC T

TGTCTGGTGC

GCACCTCTA

AGGAtAG'"-- .ACCTGAAGC-1 ATTCCT TOCT

TAGAAGTTGG

ATGTTCTTTA

GCATATTTTA

GCACAGCGCT CGGACTTACG ATATGGIGGCC GAGCTTGCCC GGZAGCCAATC AAGGAACAGT TGTGGTGGCT GTTGAAGTGA CAAGCCTGAG CCTCCAGAAG GAGGGATGTT TT'TGGTCACA CCACACCTTG GGGAGATGCC CCCCCTTA'TC TTTGACAACT GCTTCAGTTG CCATCTGATA

GAAGT.ATGCA.CAGGACGAAG

TTCr-TGAACTTIGGGTTTGCTG AGATAGCTTC GTTTTGTATT TGTTATGCGC CATAGTGATA AAATGA ACAT TGAATACAAC ATCGAATGCT TCGTTCCTGT INFORMATION FOR SEQ ID NO:86: SEQUENCE CHARACTERISTICS: LENGTH: 1410 base pairs TYPE: nucleic acid STRANDEDNESS: singie TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:86: GAAGATGGGG CTGTGGGTGG TGCTGGCTTT

GCTTACAATG

120

ATGAATTACT

180

CTGTTGTACA

240

TGTGCTGTGG

300

GAAAAGGACA

360

GAAGCCGTGG

420

GCCAGAGATG

480

GATGGACGGA

540

ATCTCCACTG

600

CTGCTGCGGGG

660

TCGTAAGTTC

ATGGGGACTC

AAAGACACAA

AGTCATGTGA

CTGACAGGAG

AGAGGGAGTG

GCGTTGTATC

AGAGCAGAGC

TTCTGTCTCG

CTCACAGCGT

AAGCAATCCT

TTGCCCT.CAG

GAACACTGCA

TGCATCGCTT

CTTCGGCCTC

CCCCGGGGTC

GTTGGGAGGA

AGATGTCGTG

CTTCAAAGCC

GGGGAGGAi'CT

GGCGCTCAGT

ACTGGGAGTT

GCTGAAGAGA

TTCTCATGGC

CTGTTGGACT

CGCAACTTTA

GTTTCCTGTG

CCATACATTC

GAGAATTACC

ATGGGAATCG

CACTGCGTGA

GCGCACTATT

ACAGTGAGAA

TCATTGCTGA

TTAGAAATAT

CAACAAGGAA

GGTATTTGGA

CAGATATACT

CCCTGAAGAC

TGCCCOATCA

ACACC'CGTGG

AGCTGGTGCA

GCAGTCTCAG

TGGATTGGTG

ACAAGTACGC

TTTCCATGAC

CAGCATATCA

TACCATCAAG

CGTTCTCTCT

GGGAAGAAGA

CAATGAGAGC

GGTTGTTGCA

CAGGCTGTAC

WO 98/11205 CCCG:_ AGT AG 720

CACGCCATCC

780

AAOCTGCACA

840

CAGCAACTGT

900

GAATACTTCT

960

ACCGCGCTC

1020

AGCAAGCCTT

1080

TGGTGGGCAT

1140 GO TOTGAC TA_ 1200 CT TAT TC CCT 1260

ATTCTAGTAT

1320

AAATATGACA

1380

GACAACTACA

1410

ATCCGACACT

C CAAC C CGAA

ACAACTACTA

ATGCAGATTC

TCAAATACTT

GAGGOAO.AAAT

GAGCGATAC

TTCATATAT.A

TG CCC TGC A

TATGTAACCA

AATTT TOTCA

ACTACCTATC

TATATTCTTT

COACCCT CCC

GGCACTGCAG

COT CAACCTC

CACOACCACO

CTCCCCOOCCG

C C T CGGCAC T CAAP T GC CG C AAT TCAOTT AT CACATC CA

GAACCTTTTA

TCAG-1TAAAC

TTCTTT-GCTC

A~aArAAAAAA C ACG CT G.;CC

TATGTCCA

ATGAACAACA

CCC TAT OlGA

CTCACCATCC

TCTCCCTC A

AGTGOTOCCA

TO GCGT T T TTA T GAA CCnC AA.

T T ATAAG CA.A.

TTCCCTCC~rT ATCTG1ATAAT ACAI.TGAGOC A ACOAC 02000 AGGGOCC CT

AGPACATGCC

T_-CTTAGAA

AAAACAAT

GTGATACOT

TTACA'AAP_!TC

CC OAAC CCT C

A.AACACAAT

CATTACT

AACCCCAAAC

PCTINZ97/00112 £AAO;TOC CCS AAT7-AOTCCAC ;A:AAA CCaOG

CAATCCT=T

S A.TGC ACAAG CCT CTCTCTT GA AAC SnCTA -TAP~-~1 INFORMATION -FOR SEQ ID NO:37: SEQUENCE CHARACTERISTICS: LENGTH: 687 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:87: CTAOTTTCCT TTTACAACAA TC-TCAOOTTT TGAATCTCAC AATAO:TOCG AAAOAGAAC

ATGACGAAGT

120

ATA-ATTTCTCG

180 CTTT COT CA 240 CT T CACCC 300

CATCACTOTT

360

CCCAOTCAOC

420

OACGAP±ATTA

480

CTOCCTTTCO

540

COCCOCACAC

600

CCAACTTTAA

660

GAAATGOTTG

687 ACOT CAT CGCT

TCAATCGATT

CATTTTATAA

COTTGCACCA

TTOTGCAOOC

AACAOGCTCA

AAACCGCTGT

CT CCTCGT CA

ATAGCCTAAA

ATTTOACACA

CTCTTCAGCT

TAOCTCCATT

ACT TGTCCAT

OOACAOTTOC

AOATATCACT

TTOCGATOCC3

GCCAAACTTA

AGAAGCTAGC

CT'_CTCCOC

OTTTCGCCACT

OCTCAT GAAC

OCCACAC

GTOTG '7'

OAAGATCUATC

CCCGACTTCCG

CAGCCOCAC

TCCGT OTTCC

ACACTAAGAG

TOCAOTGOOC

TCAGCAOGCC

CAATCCCTAG

ATTTTTCCCT

"nG:.T CITT

GCT

TCAAkAGCC

AOCCATAOT

CCTTCCTOAO

TGACAGGAAC

CCCOOCCCTT

ITCGTAACTTG

CA-AAATTTCC

TTCTCSCCAA

CCAAAGGATT

TTCTGCCO TGCC ITO GCAT CCC

GCA-AATCGGTA

ACTTCATTTC

TAAAAGAAAC

CCAOCTOAT C

TCCACACAIT

ACTACCACT

TATACCAACT

CAG TITT CC INFORMATION FOR SEQ ID NO:88: SEQUENCE CHARACTE'7RISTICS: WO 98/11205 WO 9811205PCT/NZ97/OO1 12 LENGTH: 683 base pairs TYPE: nuciei;c acid STRANDEDNESS: single TOPOLOGY: ILinear (xi)

GTAGTTTCGT

GATGACGA.AG

120

CATAATTTC:-

180

GCTTTCGTGG

240 ACTTGAGCrC2- 300

CATGACTGT:-

360

CCCC'UAGTGI

420 T CGACGAAPJ7 480 TTrTGGC- 540

TGGCCGCAGA;

600

TCCAACTTTA.

660

CGAAATGGTT

688 SEQUENCE DESCRIPTION: SEQ 1D NO:88: TTTACAACA.A TCTACAGGTT TTCA.;ATCTCA GAATAGTTGC CA;AAGGAAIGC

TPCGTGATCG

GTCAATGGAT

ACAT-T-TATA

GCGTTGGACG

TT GTGCAGGG

GCAACAGGCT

T AAAC CC T G GCTG C TCGCT

GATAGCCTAA

AATTT'GACAC

GCTCTTCAGG

TTAGCTCCAT

TAGTTGTCCA

AGGACAGTTG

AAGATAT CAC 77TSCGM'TGGGj CAG CCAAACT

GTAGAAGCTA

GACTCC-rGTCG

AGTTTGCCAG

AGCTGATGAA

TGGCACAC

T G T rGT TT C

TGAAGATGAT

COCCGACTT'3 TC a GG CC GC A T C C'-7G

TFAACACTAAG

G C T SC -GTG G

CTC:AGGAGGC

T C~ T CCG T A

CATTTTTGGC

TTTGTATTTG

CTGTCAAAGC

GAGOCCATAG

GGTrTGCTGAG TCACAG GAAC AGCCC GGG CC

GGTTGTAACT

CCAABT-AATTTC-

GTTCTCGCCA

TC CAAAGGAT

:T--CTGCGTG

C:GSTGCA TGG

::GCAGCTGA

7 -:TGCaC GA.CA

AGATACC.MAC

7'CAGTTTGGC

Claims (50)

1. An isolated DNA sequence comprising a nucleotide sequence selected from the group consisting of sequences recited in SEQ ID NO: 12 and 57; complements of the sequences recited in SEQ ID NO: 12 and 57; reverse complements of the sequences recited in SEQ ID NO: 12 and 57; and reverse sequences of the sequences recited in SEQ ID NO: 12 and 57.

2. A DNA construct comprising a DNA sequence according to claim 1.

3. A transgenic cell comprising a DNA construct according to claim 2.

4. A DNA construct comprising, in the direction: ooo° a gene promoter sequence, an open reading frame coding for an enzyme encoded by a oleo nucleotide sequence selected from the group consisting of sequences recited in SEQ ID NO: 12 and 57 and sequences having at least about a 99% probability of being the same as a sequence of SEQ ID NO: 12 and 57 as measured by computer algorithm FASTA; and a gene termination sequence. *o. The DNA construct of claim 4 wherein the open reading frame is in a sense orientation.

6. The DNA construct of claim 4 wherein the open reading frame is in an antisense orientation. 7/03/01 73

7. The DNA construct of claim 4, wherein the gene promoter sequence and gene termination sequences are functional in a plant host.

8. The DNA construct of claim 4, wherein the gene promoter sequence provides for transcription in xylem.

9. The DNA construct of claim 4 further comprising a marker for identification of transformed cells. A DNA construct comprising, in the direction: a gene promoter sequence, a non-coding region of a gene coding for an enzyme encoded by a nucleotide sequence selected from the group consisting of sequences recited in SEQ ID NO: 12 and 57 and sequences having at least about a 99% probability of being the same as a sequence of SEQ ID NO: 12 and 57 as measured by computer algorithm FASTA; and i" a gene termination sequence. loll e

11. The DNA construct of claim 10 wherein the non-coding region is in a sense orientation C..

12. The DNA construct of claim 10 wherein the non-coding region is in an antisense orientation. CO C l 13. The DNA construct of claim 10, wherein the gene promoter sequence and gene termination sequences are functional in a plant host.

14. The DNA construct of claim 10, wherein the gene promoter sequence provides for transcription in xylem. A transgenic plant cell comprising a DNA construct, the DNA construct comprising, in the 5' 3' direction: 7/03/01 74 a gene promoter sequence: an open reading frame coding for an enzyme encoded by a nucleotide sequence selected from the group consisting of sequences recited in SEQ ID NO: 12 and 57 and sequences having at least about a 99% probability of being the same as a sequence of SEQ ID NO: 12 and 57 as measured by computer algorithm FASTA; and a gene termination sequence.

16. The transgenic plant cell of claim 15 wherein the open reading frame is in a sense orientation.

17. The transgenic plant cell of claim 15 wherein the open reading frame is in an antisense orientation.

18. The transgenic plant cell of claim 15 wherein the DNA construct further comprises a marker for identification of transformed cells. *ao

19. A plant comprising a transgenic plant cell according to claim 15, or fruit or seeds thereof.

20. The plant of claim 19 wherein the plant is a woody plant. ee

21. The plant of claim 20 wherein the plant is selected from the group :0o. consisting of eucalyptus and pine species. l

22. A transgenic plant cell comprising a DNA construct, the DNA construct comprising, in the 5' 3' direction: a gene promoter sequence; a non-coding region of a gene coding for an enzyme encoded by a nucleotide sequence selected from the group consisting of sequences recited in SEQ ID NO: 12 and 57 and sequences having at least about a 7/03/01 99% probability of being the same as a sequence of SEQ ID NO: 12 and 57 as measured by computer algorithm FASTA; and a gene termination sequence.

23. The transgenic plant cell of claim 22 wherein the non-coding region is in a sense orientation.

24. The transgenic plant cell of claim 22 wherein the non-coding region is in an antisense orientation. A plant comprising a transgenic plant cell according to claim 22, or fruit or seeds thereof.

26. The plant of claim 25 wherein the plant is a woody plant.

27. The plant of claim 26, wherein the plant is selected from the group consisting of eucalyptus and pine species.

28. A method for modulating the lignin content of a plant comprising stably incorporating into the genome of the plant a DNA construct comprising, in the direction: •g a gene promoter sequence; o an open reading frame coding for an enzyme encoded by a nucleotide sequence selected from the group consisting of sequences recited in SEQ ID NO: 12 and 57 and sequences having at least about a 99% probability of being the same as a sequence of SEQ ID NO: 12 and 57 as measured by computer algorithm FASTA; and a gene termination sequence.

29. The method of claim 28 wherein the plant is selected from the group _consisting of eucalyptus and pine species. 7/03/01 76 The method of claim 28 wherein the open reading frame is in a sense orientation.

31. The method of claim 28 wherein the open reading frame is in an antisense orientation.

32. A method for modulating the lignin content of a plant comprising stably incorporating into the genome of the plant a DNA construct comprising, in the direction: a gene promoter sequence; a non-coding region of a gene coding for an enzyme encoded by a nucleotide sequence selected from the group consisting of sequences recited in SEQ ID NO: 12 and 57 and sequences having at least about a 99% probability of being the same as a sequence of SEQ ID NO: 12 and 57 as measured by computer algorithm FASTA; and a a gene termination sequence. a

33. The method of claim 32 wherein the non-coding regioh is in a sense _.orientation. oo••

34. The method of claim 32 wherein the non-coding region is in an i antisense orientation. :35. The method of claim 32 wherein the plant is a woody plant. a.

36. The method of claim 35, wherein the plant is selected from the group consisting of eucalyptus and pine species.

37. A method for producing a plant having altered lignin structure comprising: transforming a plant cell with a DNA construct comprising, in the direction, a gene promoter sequence, an open reading frame 7/03/01 coding for an enzyme encoded by a nucleotide sequence selected from the group consisting of sequences recited in SEQ ID NO: 12 and 57 and sequences having at least about a 99% probability of being the same as a sequence of SEQ ID NO: 12 and 57 as measured by computer algorithm FASTA, and a gene termination sequence to provide a transgenic cell; cultivating the transgenic cell under conditions conducive to regeneration and mature plant growth.

38. The method of claim 37 wherein the open reading frame is in a sense orientation.

39. The method of claim 37 wherein the open reading frame is in an antisense orientation. The method of claim 37 wherein the plant is a woody plant.

41. The method of claim 40 wherein the plant is selected from the group consisting of eucalyptus and pine species.

42. A method for producing a plant having altered lignin structure comprising: transforming a plant cell with a DNA construct comprising, in the direction, a gene promoter sequence, a non-coding region of a 0o gene coding for an enzyme encoded by nucleotide sequence selected from the group consisting of sequences recited SEQ ID NO: 12 and 57 and sequences having at least about a 99% probability of being the same as a sequence of SEQ ID NO: 12 and 57 as measured by computer algorithm FASTA, and a gene termination sequence to provide a transgenic cell; cultivating the transgenic cell under conditions conducive to regeneration and mature plant growth. 7/03/01

43. The method of claim 42 wherein the non-coding region is in a sense orientation.

44. The method of claim 42 wherein the non-coding region is in an antisense orientation. The method of claim 42 wherein the plant is a woody plant.

46. The method of claim 45 wherein the plant is selected from the group consisting of eucalyptus and pine species.

47. A method of modifying the activity of an enzyme in a plant comprising stably incorporating into the genome of the plant a DNA construct including: a gene promoter sequence; an open reading frame coding for an enzyme encoded by a nucleotide sequence selected from the group consisting of sequences *ebb recited in SEQ ID NO: 12 and 57 and sequences having at least about a 0:0 99% probability of being the same as a sequence of SEQ ID NO: 12 and 57 as measured by computer algorithm FASTA; and a gene termination sequence. oo•°

48. The method of claim 47 wherein the open reading frame is in a sense orientation. 0*

49. The method of claim 47 wherein the open reading frame is in an antisense orientation. A method of modifying the activity of an enzyme in a plant comprising stably incorporating into the genome of the plant a DNA construct including: a gene promoter sequence; 7/03/01 09 a 0 09*: @0 0 0*00 8000 s 0 3 0 *0*0 0 0800 0 0 0 06 0 0 S 0 a non-coding region of a gene coding for an enzyme encoded by a nucleotide sequence selected from the group consisting of sequences recited in SEQ ID NO: 12 and 57 and sequences having at least about a 99% probability of being the same as a sequence of SEQ ID NO: 12 and 57 as measured by computer algorithm FASTA; and a gene termination sequence.

51. The method of claim 50 wherein the non-coding region is in a sense orientation.

52. The method of claim 50 wherein the non-coding region is in an antisense orientation.

53. The method of claim 50 wherein the plant is a woody plant.

54. The method of claim 53 wherein the plant is selected from the group consisting of eucalyptus and pine species. An isolated DNA sequence comprising a nucleotide sequence having at least about a 99% probability of being the same as a sequence of SEQ ID NO: 12 and 57 as measured by computer algorithm FASTA.

56. A DNA construct comprising a DNA sequence of claim

57. A transgenic plant comprising a transgenic cell of claim 3.

58. A method for producing a plant having modified lignin structure comprising: transforming a plant cell with a DNA construct of claim 3 to provide a transgenic cell; and cultivating the transgenic cell under conditions conducive to regeneration and mature plant growth. 7/03/01

59. A method for modifying the activity of an enzyme in a plant comprising stably incorporating into the genome of the plant a DNA construct of claim 3. An isolated DNA sequence comprising a nucleotide sequence having at least 90% identity to a sequence of SEQ ID NO: 12 or 57 as measured by computer algorithm FASTA.

61. A DNA construct comprising a DNA sequence according to claim

62. A transgenic cell comprising a DNA construct according to claim 61. Dated this 7th day of March, 2001 Genesis Research Development Corporation Limited :and Fletcher Challenge Forests Limited Patent Attorneys for the Applicant *PETER MAXWELL ASSOCIATES o**o 0* o *oo *oooo *oooo* *ooo *oo OoO o *go• 7/03/01