MAMMARY GLAND-SPECIFIC PROMOTERS

Abstract: An isolated DNA fragment comprising a mammary gland-specific promoter is disclosed. Preferably, this promoter promotes gene expression throughout the estrous cycle in a constant manner. In one embodiment, the promoter comprises nucleotides 1154 through 2967 of SEQ ID NO:1 or 1102 through 2910 of SEQ ID NO:2.

Description: MAMMARY GLAND-SPECIFIC PROMOTERS Field Of The Invention In general, the field of the invention is mammary gland- specific promoters. Specifically, the field of the present invention is mammary gland-specific promoters isolated from the rat NRL gene and the human NGAL gene . Background The oncogene c-erbB-2 is known to be associated with the clinical progression of human breast cancer. In vi vo models utilizing c-erbB-2 ' s rodent homolog, neu , have been developed to try to evaluate the role of c-erbB-2 in mammary carcinogenesis and tumor biology. In one model, transgenic mice have been generated in which the expression of activated neu is targeted to the mammary gland using mammary-specific promoters. In a second model the activated neu oncogene has been directly and stably introduced into in si tu rat mammary epithelial cells, using a replication-defective retroviral vector. With both methods, neu was found to be a potent tumor inducer. The isolation of a lipocalin uniquely overexpressed in neu- initiated rat mammary carcinomas has been reported in S. Stoesz, et al . , 1994 AACR Abstract. This lipocalin has been named "NRL" for neu-related lipocalin. (The disclosure of this abstract and of all other publications referred to herein are incorporated by reference as if fully set forth herein.) As lipocalins are known to have a wide range of functions, the specific function of NRL is not known. A protein somewhat homologous to rat NRL, human NGAL, has been isolated and sequenced. Various cDNA gene sequences coding for NGAL and the NGAL protein sequence have been reported in L. Kjeldsen, et al . , J. Biol . Chem . 268:10425-10432 (1993); J. Bundgaard, et al . , Biochem. Biophys . Res. Comm. 202 [3] : 1468-1475 (1994); S. Bartsch, et al . , FEES Let . 37:255-289 (1995). NGAL (also known as neutrophil lipocalin/HNL) has been found in a variety of cell types (e.g. bone marrow; ovarian cell cancers) . Again, the specific function of NGAL is not known. Note that Bundgaard, et al . reported the first base of the mature protein as Q from CAG, whereas Kjeldsen, et al . at one location reported an E at that position. The present claims use "NGAL" to cover both variants. Summary Of The Invention In one embodiment, the present invention is an isolated DNA fragment comprising a mammary gland-specific promoter that promotes gene expression throughout the reproductive cycle. In a preferred embodiment of the invention, the promoter promotes gene expression in a constant manner throughout the estrous cycle. The promoter also promotes gene expression throughout the pregnancy, lactation and involution stages in a non-constant manner. The promoter is capable of strongest gene promotion in the involution stage and the weakest gene promotion in the pregnancy stage. In one particularly advantageous form of the invention, the promoter is the rat NRL promoter or the human NGAL promoter. However, promoters to homologs of the NRL and NGAL genes found in other species are also envisioned to be suitable for the present invention. For example, an NGAL homolog has been isolated from mouse (24P3 oncoprotein) . In one embodiment, the mammary gland-specific promoter comprises nucleotides 1 through 2967 of SEQ ID NO : 1. Preferably, the promoter comprises nucleotide sequences 1154 through 2967 of SEQ ID NO : 1. In another embodiment of the present invention, the promoter comprises sequences 1 through 2910 of SEQ ID NO : 2. Preferably, the promoter comprises nucleotides 1102 through 2910 of SEQ ID NO: 2. In another embodiment of the invention, the promoter consists of fragments of SEQ ID NOs : 1 and 2 that have been truncated at the 5' -end. These truncated fragments will still possess the same mammary-specific gene expression properties as full-length SEQ ID NOs : 1 and 2. In another embodiment of the present invention, the promoter is part of a vector molecule, such as a plasmid or virus vector. In an especially preferred embodiment, the vector molecule comprises a mammary gland-specific promoter operably connected to a gene sequence. This embodiment of the present invention may be used to create a transgenic animal with gene expression targeted to the mammary gland in a developmental manner consistent with the expression of the rat NRL and human NGAL genes . In another embodiment, the present invention is a transgenic non-human mammal comprising the vector described above . It is an object of the present invention to provide a mammary gland-specific promoter with distinctive developmental regulation. It is another object of the present invention to provide a mammary gland promoter with constant expression throughout the estrous cycle. It is another object of the present invention to provide a transgenic mammal containing the promoter of the present invention operably connected to a foreign gene. It is another object of the present invention to provide a mammary-specific promoter as a drug target for breast cancers. Other objects, features and advantages of the present invention will become apparent after examination of the specification, drawings and claims. Description Of The Drawings Fig. 1 is a sequence comparison of cDNAs of hPGDS2, rPGDS2 , rNRL and hNGAL. Fig. 2 is a sequence analysis of rat NRL promoter (SEQ ID N0:1) . Fig. 3 is a sequence analysis of human NGAL promoter (SEQ ID NO: 2) . Fig. 4 is a sequence comparison between rat NRL and human NGAL promoters. Description Of The Invention In one embodiment, the present invention is an isolated DNA fragment comprising a mammary gland-specific promoter capable of promoting gene expression throughout the mammalian reproductive cycle. The promoter is most conveniently isolated from the 5' region of either the rat NRL gene or the human NGAL gene. The Examples below demonstrate one particularly advantageous method of isolating the promoter. The mouse mammary tumor virus (MMTV) promoter (Truss, et al . , J. Steroid Biochem . Molec . Biol . 43 [5] 365-378 , 1992) and the whey acidic protein (WAP) gene promoter (Li and Rosen, Mol ecular Endrochrinology , 8 [10] : 1328-1334 , 1994) are two examples of mammary-specific promoters with developmental expression patterns that differ from the promoters of the present invention. In the method described below, specific PCR promoters were developed from a comparison of the rat NRL cDNA, the human NGAL cDNA and the rat and human brain prostaglandin D2 synthase cDNA (see Fig. 1) . Fig. 1 is a sequence comparison of cDNAs of hPGDS2, rPGDS2, rNRL and hNGAL (SEQ ID NOs : 3 , 4, 5 and 6) . The inverted triangles indicate the splicing sites that have been characterized for human prostaglandin D 2 synthase gene. Primers rNRL-A2, hNGAL-A2, rNRL-R3 and hNGAL-R2 (SEQ ID NOs : 7 , 8, 9 and 10) were useful in amplifying these 5' regions. This amplification was by standard PCR reaction. In this manner, a set of genomic clones were isolated corresponding to the rat NRL gene and the human NGAL gene . If one of skill in the art is interested in isolating promoters from other NGAL homologs, such as mouse, an analogous procedure may be followed. The mouse NGAL (24p3) was initially isolated in a search for genes overexpressed during a SV40-induced mitotic reaction (Oncogene 4(g) :601-608, 1989) . The gene encodes the mouse 24p3 and promoter region (793 bp) has recently been isolated (Gene 170 (2) .173-180, 1996) . One may test these candidate promoters by the experiments described below the Examples. A successful mammary gland- specific promoter will promote gene expression in a constant manner throughout the estrous cycle. These clones were analyzed and screened for the first exon- containing clones by a standard PCR method using a first exon forward primer and a second exon reverse primer of the NRL NGAL genes, respectively. SEQ ID NOs:l and 2 (Figs. 2 and 3) are the result of sequence analysis of the rat NRL 5' region and the human NGAL 5' region, respectively. Fig. 2 is a sequence analysis of rat NRL promoter (SEQ ID N0:1). "+1" indicates the putative transcription start site. The promoter and upstream region is indicated in negative number relative to the transcription start site. Putative response elements are also indicated. Fig. 3 is a sequence analysis of human NGAL promoter (SEQ ID NO: 2) . " + 1" indicates the putative transcription start site. The promoter and upstream region is indicated by a negative number relative to the transcription start site. Putative response elements are also indicated. Another way to obtain the same promoter sequences would be to use primers derived from SEQ ID NOs : 1 and 2 to directly amplify the sequences found in SEQ ID NOs : 1 and 2 from rat or human genomic DNA. It is well known by those of skill in molecular biology that the entire 5' region of a gene is not needed to confer specific developmental regulation properties. We analyzed the sequences in SEQ ID NOs : 1 and 2 to determine what regions would be strictly necessary for developmental regulation consistent with the natural regulation of the rat NRL or human NGAL gene. Fig. 4 is a sequence comparison between rat NRL and human NGAL promoters. Periods are introduced for best alignment between NRL and NGAL. Vertical bars indicate the bases identical between NRL and NGAL. The regions with long stretch of homology are high-lighted with vertical bars. Tables 1 and 2 in the Examples below detail the placement of the TATA box and putative ERE and PRE/GRE half-sites. Because the two promoters demonstrate homology up to base -1810, we envision that at least 1810 nucleotides of each 5' region are needed for sufficient developmental regulation. However, we envision that further truncations from the 5' -end of the promoter will also result in promoter fragments with equivalent abilities to promote mammary- specific gene expression. These promoters are also envisioned to be suitable for the present invention. Additionally, we envision that the nucleotide region between 1154 and 2967 of SEQ ID NO:l and 1102 and 2910 of SEQ ID NO: 2 are preferred promoters of the present invention. In one embodiment, the present invention is a promoter that provides developmental expression of a gene consistent with the natural expression of the rat NRL and human NGAL genes. We have studied the expression of these RNAs in different stages of the mammalian reproductive cycle, including estrous, pregnancy, lactation and involution stages. The expression of both RNAs is relatively constant throughout the stages of the estrous cycle (diestrous, proestrous and estrous) . By "constant" we mean that the expression does not change by more than 10% throughout the estrous cycle. The mRNAs are also expressed throughout pregnancy and lactation and involution. In both systems, the mRNA is expressed most strongly in involution and most weakly in pregnancy. As the examples below demonstrate, expression varies between .5 fold in the pregnant mammary gland to 4 fold during involution. In another embodiment, the present invention is the isolated promoter fragment combined in a vector, most preferably operably connected to a foreign gene sequence. Preferable examples of vectors include both plasmid and viral vectors. In this manner, foreign gene expression can be targeted to a mammary gland. For example, one may want to target the expression of a therapeutic molecule to the estrous cycle. It is an advantage in many applications that the expression is fairly level or constant throughout the various stages of reproduction . Another embodiment of the present invention is the use of the mammary-specific promoters as a drug target for breast cancer genes. As described above, these genes are overexpressed in a subset of breast cancers and the up-stream regions of these genes are likely therapeutic targets. Examples mRNA Expression Human diseases or cancers are usually studied using animal models in order to obtain important information that may one day be applied to human therapy. Of the known oncogenes, c-erbB2 is most commonly associated with the clinical progression of human breast cancer. Activated neu oncogene (neu * ) , the rodent equivalent of c-erbB2 , is extremely potent in inducing mammary carcinoma in rats. To explore the mechanism of neu * -initiated mammary carcinogenesis, a subtraction hybridization-based method was used to isolate cDNA clones derived from mRNAs that were differentially expressed in πeu * -induced tumors versus normal mammary glands. One cDNA clone was isolated and designated as neu-related lipocalin NRL (Stoesz and Gould, supra , 1995) . Northern analysis using NRL cDNA probe revealed that NRL mRNA levels in πeu * -induced tumors is 12-fold of that in normal mammary glands. This enhanced expression is specific to activated neu-induced tumors and is not observed in tumors induced either by activated-ras, chemical carcinogens DMBA, or NMU. Sequence analysis revealed that rat NRL is highly homologous to the human neutrophil gelatinase associated lipocalin (NGAL) and mouse oncogenic lipocalin-24P3. A tissue distribution study revealed that expression of NRL mRNA was largely confined to the mammary gland (Stoesz and Gould, supra, 1995) . No mRNA expression was observed in rat liver, spleen, muscle, kidney or brain. The expression of NRL in the mammary gland under different physiological conditions was evaluated by Northern blot analysis. Expression of the NRL mRNA did not vary significantly throughout the estrous cycle, but varied significantly during pregnancy (day 18), lactation (day 4) and involution (day 6) . The mRNA levels decreased in the pregnant mammary gland (0.5-fold expression compared to virgin mammary gland) , then increased during the lactation (1.5- fold) and involution (> 4 -fold) . The expression of the gene only in mammary gland and steady mRNA levels during estrous cycle suggest that the NRL gene has a mammary gland-specific promoter than is not significantly regulated by estrous hormones. The increased mRNA levels of NRL during involution suggests that NRL expression may be directly or indirectly regulated by estrogen. A preliminary study with a human specimen revealed that the NGAL (human homologue of NRL) expression levels were inversely correlated with estrogen receptor levels. In the Examples below, we describe the cloning, sequencing and sequence analyses of promoters and up-stream regions of the rat NRL and human NGAL genes . Genomic Clone Isolation There are several different ways to isolate a promoter sequence. One approach is to screen a genomic library using a cDNA probe corresponding to the gene studied. This approach is fairly labor-intensive and does not always generate clones containing the promoter and up-stream transcriptional regulatory sequences. The second approach is to amplify the pro oter- containing fragment directly from genomic DNA by a PCR-based method. This approach requires cDNA sequence of the very first exon of the gene studied and each run only generates a few hundred bases new sequence. Several runs of extension may be necessary to cover a range of few thousand base pairs and mis- incorporation of nucleotide could be introduced by PCR during the repeated process. The third approach, which we took, is to PCR-amplify genomic fragments using oligonucleotide primers derived from rat NRL and human NGAL cDNA sequences. From the sequences of genomic fragments, we generate genomic specific oligonucleotide primers and use them to isolate PI plasmid clones by PCR screening. The average insert size of a PI clone is about 80 to 100 kb, which is large enough to cover an entire gene of average size. The PI clone can serve not only as a source of promoter sequence but also a good probe for chromosomal localization by fluorescence in si tu hybridization. Gene structure of either the rat NRL or the human NGAL is not known. It has been observed that genes within the same family, especially within the same subfamily, usually share a common gene structure. Therefore, we compared the cDNA sequences of rat NRL and human NGAL to the rat and human brain prostaglandin D 2 synthetase cDNAs that also belong to the lipocalin family with known intron-exon boundaries. Although a relatively small transcript, the human brain prostaglandin D 2 synthetase gene contains 7 exons (White, et al . J. Biol . Chem . 267 (32) :23202-23208, 1992). The sequences spanning the intron- exon boundaries are well conserved among rat PGDS2 , human PGDS2 , rat NRA and human NGAL cDNAs (Fig. 1, SEQ ID NOs : 3, 4, 5 and 6) . Several primer sets were prepared and tested for PCR- amplification of NRL from rat spleen DNA and NGAL from human DNA (MCF-7 cells) . Genomic fragments of rat NRL and human NGAL were amplified by PCR with primers derived from the second and the third exons . Partial sequencing of the amplified NRL and NGAL DNA revealed the intron-exon boundaries at exactly the position predicted from Fig. 1. In combination with forward primers (rNRL-A2 and hNGAL-A2, SEQ ID NOs : 7 and 8) of the second exon, reverse primers (rNRL-R3 and hNGAL-R2, SEQ ID NOs : 9 and 10) derived from the second intron were tested for PCR amplification. Genomic fragments of predicted sizes were amplified and sequences determined. These characterized rat NRL primer set (rNRL-A2 and rNRL-R3) and human NGAL set (hNGAL-A2 and hNGAL-R2) were sent to Genome Systems St. Louis, Missouri, to isolate the corresponding PI clones by a PCR-based method. In this manner, three independent PI plasmid clones were isolated for rat NRL and three for human NGAL. Host cells harboring the PI plasmid were further verified by PCR using rNRL and hHNGAL primer sets that differ from those used for screening. The verified PI clones were then purified by re- plating the cells at low density and examined by PCR. PI plasmid preparations of purified clones were digested with either endonucleases EcoRl or Hindlll and subjected to Southern blot analysis with DNA probe containing the first exon. The rat NRL probe detected a 8-kb EcoRl band and a 3-kb Hindlll band of NRL PI plasmid respectively. The human hNGAL probe only hybridized to bands of relatively high molecular weight (>7 kb) in either digestion. Aliquots of the same digestions used for Southern analysis were subcloned into modified pSP73 plasmid vector (with a Notl site introduced at Smal site) . The subclones were screened for first exon-containing clones by a PCR method using a first-exon forward primer and second-exon reverse primer of the NRL and NGAL genes, respectively. Plasmid DNAs of positive clones were mapped by restriction digestion and PCR amplification with various primer combinations of NRL, NGAL and vector primers. Selected positive clones were then sequenced upward from the end of the first exon and downward from the 5' -end. Both rat NRL and human NGAL sequences were analyzed for putative transcription regulatory elements . Fig. 2 is a sequence 5' region of the region of the rat NRL gene. SEQ ID NO:l repeats this sequence. Fig. 3 is the sequence 5' region of the human NGAL gene. SEQ ID NO: 2 repeat this sequence. Analysis Of The 5' -end Of NRL And NGAL Genes Table 1, below summarizes some of our analysis of the 5' up stream region of the NRL and NGAL genes. The "5' up-stream region" designated in the table extends from the beginning of the 5 '-end of the sequenced region to the putative transcription initiation site. The TATA-box designation is relative to the putative transcription initiation at +1 in Figs. 2 and 3 for the rat NRL gene and for the human NGAL gene. Referring to Table 1, the TATA box is where the general transcription machinery recognizes and binds. It has been observed that the NGAL levels are inversely correlated to the estrogen receptor (ER) and/or progesterone receptor (PR) levels in T47D cell (a human breast cancer cell line) . ER or PR regulate their target gene expression through binding to those specific response elements (ERE or PRE) in the regulatory regions. Therefore, searching for putative estrogen receptor binding sites or progesterone receptor bind site will facilitate characterizing the role of ER or PR in NGAL expression. Negative GRE/PREs have been identified in bovine prolactin promoter ( Gene & Development 2:1144-1154, 1988) and rat Pro-Opiomelanocortin promoter (Mol . Cell. Biol . 9(12), 1989) . Negative ERE may contain sequence similar to that of positive ERE which consists of direct repeats of A / G GGTCA half -site. Table 1 Rat NRL genomic DNA Human NGAL genomic DNA 5' upstream region 2967 bp 2910 bp TATA-box ATAAAGA at -29 ATAAATA at -29 putative ERE-half site A/QGGTCA, many A/ G GGTCA, many putative-(PRE/GRE) TCYACNnnnTGATCW, many TCYACNnnnTGATCW, many The rat NRL and human NGAL promoters are highly homologous to each other in the regions listed above. Figure 4 is a sequence comparison between the rat NRL and the human NGAL promoters. No extended homology was identified between rat NRL and human NGAL beyond the base -1820. Response elements identified within the homologous regions include TATA box, nuclear factor 1 (NF-1) -binding site, NF-KB-binding site, negative glucocorticoid/progesterone response element (nGRE/nPRE) , and a half-site of estrogen response element (1/2 ERE) . Additional elements yet to be identified are also presented in these homologous regions. These homologous regions likely harbor those transcription regulatory elements important to both rat NRL and human NGAL genes. Therefore, these regions may also be conserved in NRL/NGAL homologs of all other mammalian species as well. Table 2 Regions with extended homology NRL promoter NGAL promoter Putative response elements -32 to +18 -32 to +22 TATA box -151 to -56 -127 to -34 NF-1 binding site, NF-κB-bιndιng site -192 to -171 -152 to -131 -275 to -224 -217 to -167 NF-κB -binding site -462 to -370 -413 to -321 -621 to -595 -586 to -562 -651 to -635 -603 to -587 Oct-1, Oct-2 binding site -692 to -659 -634 to -601 -862 to -844 -769 to -751 -892 to -863 -806 to -777 -1027 to -1001 -844 to -818 -1088 to -1073 -914 to -899 -1133 to -1114 -959 to -940 -1243 to -1214 -1063 to -1036 nGRE/nPRE (POMC), 1/2 ERE -1529 to -1475 -1317 to -1261 -1584 to -1564 -1375 to -1355 -1641 to -1604 -1442 to -1405 nGRE/nPRE (POMC/Prolactin), 1/2 ERE -1695 to -1682 -1495 to -1482 -1810 to -1755 -1791 to -1739 We did an extensive sequence comparison of the human NGAL and rat NRL promotors to those of other members of lipocalin superfamily with known gene structure, including mouse 24p3 (X81627, Gene 170 (2) : 173-180 , 1996), human prostaglandin D 2 synthase (M98537, J. Biol . Chem. 267 (32) : 23202-23208 , 1992), rat prostaglandin D synthase (M94134, Proc . Natl . Acad . Sci . USA 89 (12) :5376-5380, 1992), human tear prealbumin (TP) gene (L14927, Gene 139 (2 ): 177-183 , 1994), rat epididylma secretory protein I (X59831, Bioche . J. 281 (Pt 1):203-210, 1992), rat von Ebner's gland protein I (X74805) and II (X74807, Eur. J. Biochem . 211(3) : 905-916, 1994) . Based on the comparison, we obtained a putative transcriptional start site for the rat NRL promotor and the human NGAL promotor. In mouse NGAL (24p3), a putative glucocorticoid response element has been identified at -520 to -501. However, the two GRE half-sites are not well conserved in human and rat NGAL promoters. Transcriptional activity analysis of serial deleted 24p3 promoter reveals that the putative GRE plays only a minor role in response to glucocorticoid (dexamethasone, Dex) . The major element responds to Dex is located between base -198 and - 155 with no homology to any known transcription factor-binding site. This region is highly homologous between mouse and rat (base -208 to -165) , but with low homology to human NGAL, suggesting that this element is rodent-specific. In summary, the characterization of mouse 24p3 promoter is still preliminary at this point. Two putative NF-κB-binding sites have been identified in the proximal region of NGAL promoters of human, rat (Table 2) and mouse. The distal NF-zB-binding site (GGGAATGTCC, SEQ ID NO: 14) is identical among human, rat and mouse (at -176, -235, - 230, respectively). The proximal NF-/cB-binding site is also conserved among these species with one or two bases variation. human GGCAATTGCC at -87 (SEQ ID NO: 11) rat GGCAATTAAC at -111 (SEQ ID NO: 12) mouse GGCAATTACT at -109 (SEQ ID NO: 13) NF-KB has been implicated in the induction of several genes involved in the early process of immune and inflammatory responses. One of those gene involved is interleukin 6 (IL-6) . Glucocorticoids are well known for their anti-inflammatory activity. Glucocorticoids strongly repress IL-6 gene expression through the direct interaction between glucocorticoid receptor and the p65 subunit of transcription factor NF-KB ( PNAS 91(2) :752-756 , 1994). In addition, estrogen receptor has also been demonstrated to repress the IL-6 promoter through interaction with NF- nB (Mol . Cell . Biol . 15 (9) : 4971-4779 , 1995). Progesterone receptor and glucocorticoid receptor share the same DNA response element. The lower expression levels of NGAL in ER/PR positive T47D cells could be due to the repression of NGAL promoter by ER and/or PR directly or through interaction with NF-KB. Only 793 base pairs upstream the transcription start site of mouse 24p3 has been reported. Homologous sequences beyond this region have been identified between human and rat NGAL promoters up to base -1810 (Table 2 and Fig. 4) . Homologous sequences in this region (-790 to -1810) include several putative negative GRE/PRE and ERE half-sites. They could be important elements involved in the regulation of NGAL promoter activity. SEQUENCE LISTING (1) GENERAL INFORMATION: (l) APPLICANT: Gould, Michael N. Chen, Kai-Shun (n) TITLE OF INVENTION: MAMMARY GLAND-SPECIFIC PROMOTERS (m) NUMBER OF SEQUENCES: 14 (IV) CORRESPONDENCE ADDRESS: (A) ADDRESSEE: Quarles & Brady (B) STREET: 411 East Wisconsin Avenue (C) CITY: Milwaukee (D) STATE: Wisconsin (E) COUNTRY: U.S.A. (F) ZIP: 53202-4497 (v) COMPUTER READABLE FORM: (A) MEDIUM TYPE: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS (D) SOFTWARE: Patent In Release #1.0, Version #1.25 (vi) CURRENT APPLICATION DATA: (A) APPLICATION NUMBER: (B) FILING DATE: (C) CLASSIFICATION: (vm) ATTORNEY/AGENT INFORMATION: (A) NAME: Baker, Jean C. (B) REGISTRATION NUMBER: 35,433 (C) REFERENCE/DOCKET NUMBER : 960296.93863 (IX) TELECOMMUNICATION INFORMATION: (A) TELEPHONE: (414) 277-5709 (B) TELEFAX: (414) 271-3552 (2) INFORMATION FOR SEQ ID NO : 1 : (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 3046 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS : double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:l: GAATTCCGCA AGCAGACCTG AGGGCCAGGC TGGAGAGTGG AGCTGCGTTC GCTCCAGCCC 60 CTCAAGGCCA GGCTCACCAG TTTCTGCAGT GAGTTTCTGG ATCAGAATGT CAGACTGGAT 120 TCTTGAAATG CAGTAACCTC GGAGCCTCTC ATGTGGAATG GACCTAGGTC GGGTTGTGTA 180 GCAGTTAGAG TTCTTGGGCT TTATGACCAC AGAAAACTCA AGTGTGACCT AGATGTGTTA 240 CTACTAAGTT CAGGGTCAGC ACAGATTACA CAATGAGACC TCATATCAAA ATAAATAATA 300 AATAATAAAA AGAAGTAGCG GGGGCTGGGG ATTTAGCTCA GTGGCAGAGC GCTTACCTAG 360 GAAGCGCAAG GCCCTGGGTT CGGTCCCCAG CTCCGAAAAA AAAAAGAAAA AAAAAAAAAA 420 AAAAAAAGAA GTGGCTGGCT TGGTTGGCGA TGTGTGCCAA CACTCAGAGG TAGAATCAAG 480 AGAACAAGGG AAGGAAGGAA GAGGGAGGAA GGAAGGGAAG GAGGGAGGAA GGAAGAAGAA 540 GGGAGGGAGG GAAGAAGGAG GTGGGAGGAA GGAAGGAAAG AAGGAAAGAG ACCGACTGGA 600 CGAGAGGTGG AGGCAGGGGG ATGAGAAGTT CAAAGTCATC TTTGGTGACA TAGGGAGTTT 660 GAGGCTACCT GGCCTTTAGG GATTCAGTTT CAGAGAGAGA GGGTTCATGG GAGAGCTGGC 720 AGGATCCTGG GGGAAGAATC AGCAGGCTGA AGGTGGCTGT GTGCCTTGTA CCTGGAACAG 780 CCAGGGTCCT GAGCTAGGCC ATCTCCCCTC CCACCCTTAA TTCTGACCTT TTAGTTTTTC 840 CAGACCCAGC TCTCTGCCCC AGTTCATACT GGCTCGGTTC CACTGGTCAC TCTGCCCCCT 900 GGTTTTCAGA CTCTAGAATA TCCTGCCTGT CCAGCTCCTC TGAGATTCTG GTCTCTGTTT 960 TTCCTGACTA AAAATTCTTG GGGGCTCTGT CTACACCCAA TAATCACCAG AGACTCAAGG 1020 GTGCCTTTGA TTTATACATG ACTTTCTTTC TTTTAAGTCA AAGGCCTTGA GTGTATCCTT 1080 TGGCTTGCTT GCCTCAAACT CCCAATGCAG ACCAGGCTCA TCTGGCCTTG AAGTCACAGA 1140 GATCCTTTGC CTCTGCACAG AGTGTTGGGC TTAAAGGTGG GAGCCACCAC ATACAGCTTT 1200 CAAGGAGACC TTTCAAGCTA ACGTGTTTAG TTGGAAGGTT GGTTCTTTGT ACTGTTGGAA 1260 ATAGAATTTG GGGCCTCCCA CGTGCTAGAC AAACCCTCCA CCATGGAGCT CTATTCCTCA 1320 GTTCTTGGAT ACCTTTTAAG GTCACAGAGG GTAGAAGGGG TGGATCCCCT AGGTCTGAGC 1380 TACAAGGGGC TGGAAGGGTG GGAGGTCCCT GGTACCTCAA GAGTGACAGG CTCTGGTGGC 1440 CACATTGTCC CCACAGCTTG GCTCAGCTTC ACTTCCTGTC CTTTCATCAT CCAGGGACCT 1500 GAGGGGACAG ATTGTAGCGC TGTAGTCTTT CTGACATGGG AGAGGGGGAA GGCTGCATCC 1560 TAGGTGTGGG GGGATGTGAG GCTATAGCCT ACTTATCAGG TTAAAATCCC CCTCTAAGCT 1620 TTCCCTCCTG GCTAACCACC CTGAGCTAAG CAGCAGTGGA AGGGGGAGGT CAGGAGCAGC 1680 AAACAGATCA ATAAGCCTTT TTAGTCCTGT GCAGGGCCAG AGGACTTCAG TCCAGCCTTA 1740 GGTCAGATGT TCAGATGCGG ATTCTGAGGA AGCCACCTGG CGGGGAGGGA AGCACAATAT 1800 AGCATCTGGG ATCCATCCAT CGCAACCTTT CAATGAATGT TAGCCAGGCC CCAGAGAGGA 1860 AAGGGCTTTT TTTTCAGCCC TAGGCTGGAA TCAGCTGGGG AGAGAAAGTC CTAAGGCTGG 1920 GGCACTAAGT TCTCCTGCTC AAGGCTATGG CCAGAGACAG GGGGATGCCT TTTCTTTTCT 1980 TTTCTTTTTT CTTTTTTTTT TTTTTTTTTT TTTTTTGGTT CTGTTTTTCG GAGCTGGGGA 2040 CCAAACCCAG GGGATGCCTT TTCAAGGGAG GATAACTTAA GGAGAAGGTG GAACCTTGCT 2100 TCTGTCCAAA GTAACTGGAG TACACTGGGC AGTTTGGACA CACACACACA CACACACACA 2160 CACACACACA CACCCCTACT TTTCCCAAGG GGCTGGTGCT CCCCCTTATC CTACGATGAC 2220 AACAAGGTTG CAAGTCCTTG CCTTTGAAAG TGGCTGTATT CTAAGGACCG TGTGGCACAG 2280 GAGAGGGGTT GTCCCTGAGA GTTCAACTGC TGCCCTGTCT GCTCCTGTAA ATGTCAGCAT 2340 GGTCATGGGA AAGCAAAGGG GCTCAAGGGA TTGGGCACCT CCAGGCTAAT CTTCTCCCTC 2400 CCTCACCCTG TGCCAGGACC AAGTCCAAGC TTGACAGGCT TGGAACAGGG TGTCCCATTC 2460 TTTCCTGTCT AAAACATTCA CTCTCCCCCG TCCTCACCTC TCCAGACAAG GAAGCTACAC 2520 AGGGTCTGGT ACAGTGAGAC AGTTCTGGTT TTCAGCAGGT GTAGGTGTGG GGCGGGGGAG 2580 GGGGGCCTTC ACCACACTCG ATGTCTTGTT TCTCATTCAC TAGGACTCCT AGAGGGTTGT 2640 GGGGGCGGGG TGGGGGTGGG AGGAAGACTG TCCAGATCTG AGCTGCTGAC CCCACAGGCA 2700 GTGCCCTTGT GCCTGCCAGA ATCCAGGGCT CTGGGAATGT CCCTTCAGAT CCCCCGTTCC 2760 CCCACCCCCC TGCAGCCCTT CCTTTTGCTC AACCTTGCAC AGTTCCTGGG GGAGAGAGGG 2820 ACAGAAATCT TGCCAAGTAT TTCAACAGGA TGTGCTGGCA ATTACCTCAT GGCTTCCTGG 2880 ACTTGGTAAA GGATGGACTA CCCCACCCTA CAAGGGGGGT TGGCAGCCAG GTAGGCCCAT 2940 AAAGAGGCCC CCTGAGGAGT CCTCCTCATT CTCTGCTCTT CCTCCTCCGG CACACATCGG 3000 ACCTAGTAGC TGCTGAAACC ATGGGCCTGG GTGTCCTGTG TCTGGC 3046 (2) INFORMATION FOR SEQ ID NO : 2 : (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 3035 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS : double (D) TOPOLOGY: linear (n) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO : 2 : AAGCTTGTGT GGTGGGACTA TGTAGAGCTG ACCCCCTCCC TGCAGCCCTG CTAGACTCTG 60 AAGAGAGCCA AGGCCAGTGG GTAGGAGGAG ACAGGTCTGG AGCTGGTGCA GAGAGAGGAA 120 TGAGCCCTGC ATGGGTTTGA TCAGAAACTC AGCCTTGTGT AGGGACACCC TGGGGCCCGG 180 TGCTGTCCAT GCATGACCTC ACAGAAGCGC AGAGCTGCCC TCTCTACAGA GGAGCGCCTG 240 ATTTGTGTGG GAGCTAGGCA GAGATCTGCA TGCATGCGGA GGAGCCAGGC TTCAAGCCAG 300 CCTGGGGGAC CCCAAGCGGG ACTATCTCCC CTTCTGCACC TGGCTCTGGT GTCTTCCCAC 360 TGTGGACCCA GTGCCCTGCT CACCCACCAC ATTCATACCC TGGAGTCCTG GGTCCTCAGA 420 GATCCATGAC ACTGCCTCAC CCCCAACTTC AAATTCTCTG GGGCTCCACC CGCTGGTCTC 480 AGCTACGTGA AGCAGTCACC GTAGACTAGA GGGTATTTTT TAGATTTAGG TCACTCTATC 540 ATCCAGGCTG GAGTGCAGTG GCACAATCAT AGCTCACTGC AGCCTCGGCT TCCTGGGCCC 600 AAGTGATCCT CCCACCTCAG CCTCCCCGAG GATACGTGGT TTTTTTTTTC TTTTTTCAGA 660 CAGGGTCTCA CTCTGTCTCC CAGGCTGGAG TGCAGTGGTG CGATCTTGGC TCACTGCAGC 720 CTCCGCCTCC CGGGTTCAAG CCATTCTCCT GCCTCAGCCT CCTGAGTAGT TGGGATCATA 780 GGCATGCATC ACCCCACCTG GCTAATTTTT GTATTTTTAG TAGAGACGGG GTTTTGCCAT 840 ATTGGCCAGG CTAGTCCCTG AGGATCATTT TTTTTTCCCC GAGATGGAGT CTCCCTCTGT 900 CGCCCAGGCT GGAGTGCAAT GGCAACCTTG GCTCACTGCA ACCTCCGCCT CCCAGGTTCA 960 AGCAATTCTT CTGCCTCAGC TTCCCGAGTA GTTGGGATTA CAGGCATGCG CCACCATGCC 1020 CAACTAATCT TTGTATTTTT ACTAGAGACA GGGTTTCACC ATGTTGGTCA GGCTGGTCTT 1080 GAACACCTGA CCTCAGGTTA TCCACCCGCC TCAGTCTCCC AAAGTGCTGG AGTTACAGGC 1140 GAGAGCCACT GCGCCCAGCC GAGGATACCT ττττττττττ TTTTAAGACA GAATATCGCT 1200 CTGTTCCAGG CTAAAGTGCA AAGGCGTGAT CTCGGCTCAC TGCAACCTCC GCCTCCCAGG 1260 TTCAAGCTGT TCTTCTGCCT CAGCCTCCCG AGTAGCTGGG ATTACAGGCG CCTGCCACCA 1320 TGCCCTGCTA ATTTTTGTAT TTTTAGTAGA GATGGGGTTT CACCGTGTTG GCCAGACTGG 1380 TCTCGAACTC CTGACCTCGT GATCCACCCG CCTCAGCCTC CCAAATGCTG GGATTACAGA 1440 TGTGAGCCAC CGCACCCGGC CTGGCAGAGG ATACTTTTTA AGGTCAAAGA CAGTAGCAGA 1500 GGTGGAGTTC CTGGGAACAG GGTCATGAGG GGAAGAGGGG GTTCGGAGGG AGCGAGTAGC 1560 CACTGGCTAC CTCTAGAAAG GGAAGGCTTT GGTGCAACAT CGTTCCCCTG CAGTTTTACT 1620 CATCTTTGCT TCCTGCCCTT TCATCATCCA ATCGGGCAGG CAGGACAGGG CCTGAGGGGG 1680 CAGGGATCCA GTGGGTGCCT CTCTAGACTA ACCCCAGCTC AGGACTCCCA GAGCCCCTTC 1740 CCTGAGGCCC TGCTGCCCCC AAGCCCAGAT TGGGGATCCC AAGCAGCACG TAGGCAGAGC 1800 CAGTGAGGTC CCCGTTAGTC CCATTGAAAG CTCTAAAACC AGCGAACCCT CAGTCCAGCC 1860 TCAGGTCAGG CATCCAGGAC GCCCTCAGCC TCATGGGTGA GCCATCTCTG CGGACACTGC 1920 ACAGGGCCTA CGATCCATCG CTGCCTCCCG AGGATGCCAG CCAGGCCCCC GTTGAGATAA 1980 CTGCTTCCCT GCTGGACAAG GCTGGGACCA GCCATCTCGG TGACAGTTCC AGAACCCCTG 2040 GCCTGGGCTG CTGGGTTCAA TGGAAAAAGG CTGTGACTAG AGTCAGGGGG ATGGTCTCAG 2100 TGACCTCAAG GATAAGGCCA GATCCTTGCA CTGTCAGTGA CCCAAAGCAA CAGGTGTCCA 2160 GAGCAGCAGT GTGGCGCCTT CACGCCCCCA CACATCAGCC CAACTCACCC AGGACAGGGA 2220 CTGTAGCCTC AGCACTCAAC CCATGTGCCC TGTGTGGGGT CTCTTCCCAC TGCACTCACA 2280 GGAGAGGAAG GGTCCCTCAG GGGTCCACTG GGGTCCCCTC CTGCAAATGG GGCAAGGAGA 2340 GGGGCAAGGG GCTGTCTCAA GGCCCCTGGA GCACATGCAG GTCCTGGACT GGGGCTCCTG 2400 GGAGGGCCAT GATTCTGGGC TCCATGAGTT CAGAGCAGAC GCCTTGTTTT TCCTTGTCCA 2460 CTGTCAGCCA CCCCACCCTT CCCTGACCCT TAAAAGAACC AGGAAACAGC ACATGATCTG 2520 TTGGAAGGAG GCATTCATTC TTTCCTTTCT GTGGGTGTGG GGAGGGACCA CAGGGCACAT 2580 ACCCCACCCT GGGATCCAGC TGAGCAGGGG GGTCAGAGAT GACAGCTCTT CCGGCTCACA 2640 GGCCACCGGC CCACATACAG GGCAATCAGA AGAAAGAAAC AGCACAAGGA AGGCACAGAG 2700 GGAGTCGTTG TCCCTGCCAG AGGTGCAGCA CTCCGGGAAT GTCCCTCACT CTCCCCGTCC 2760 CTCTGTCTTG CCCAATCCTG ACCAGGTGCA GAAATCTTGC CAAGTGTTTC CGCAGGAGTT 2820 GCTGGCAATT GCCTCACATT CCTGGCCTTG GCAAAGAATG AATCAACCCA CCCTAGATCC 2880 CATAAATAGG GCCACCCAGG TGAGCCTCTC ACTCGCCACC TCCTCTTCCA CCCCTGCCAG 2940 GCCCAGCAGC CACCACAGCG CCTCCTTCCT CGGCCCTGAA ATCATGCCCC TAGGTCTCCT 3000 GTGGCTGGGC CTAGCCCTGT TGGGGCTCTG CATGC 3035 (2) INFORMATION FOR SEQ ID NO: 3: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 827 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS : double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: CDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 3: CTCCTCCTGC ACACCTTCCG CACACCTCCC TCGCTCTCCC ACACCACTGG CACCAGGCCC 60 CGCACACCTG CTCGGCTGCA GGAGAATGGC TACTCATCAC ACGCTGTGGA TGGGACTGGT 120 CCTGCTGGGG CTGCTGGGCG GCCTACAGGC AGCACCCGAG GCCCAGGTCT CCGTGCAGCC 180 CAACTTCCAG CCGGACAAGT TCCTGGGGCG CTGGTTCAGC GCGGGCCTCG CCTCCAACTC 240 GAGCTGGCTC CAGGAGAAGA AGGCAGCGCT GTCCATGTGC AAGTCGGTGG TGGCCCCTGC 300 GGCGGATGGT GGCTTCAACC TGACCTCCAC CTTCCTCAGG AAAAACCAGT GTGAGACCCG 360 AACCATGCTG CTGCAGCCCG GGGACTCCCT CGGCTCCTAC AGCTACCGGA GTCCCCACTG 420 GGGCAGCACC TACTCTGTGT CAGTGGTGGA GACTGACTAC GACCACTACG CCCTGCTGTA 480 CAGCCAGGGC AGCAAGGGCC CCGGCGAGGA CTTCCGCATG GCCACCCTCT ACAGCCGAAC 540 CCAGACCCCC AGGGCTGAGT TAAAGGAGAA ATTTACCGCC TTCTGCAAGG CCCAGGGCTT 600 CACAGAGGAT TCCATTGTCT TCCTGCCCCA AACCGATAAG TGCATGACGG AACAATAGGA 660 CTCCCCAGAG CTGAAGCTGG GACCGCAGCC AGCCAGGTGA CCCCTGCGAT CTGGATGTTT 720 CCGCTCTGTT CCTTCCCCGA GCCCCTGCCC CGGCTCCCCG CCAAAGCACC CCTGCCCCCT 780 CGGGCTTCCT CCTGGCTCTG CGGAATAAAC TCCGGAAGCA AGTCTGT 827 (2) INFORMATION FOR SEQ ID NO: 4: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 759 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS : double (D) TOPOLOGY: linear (li) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 4: CCTCAGGCTC AGACACCTGC TCTACTCCAA GCAAATGGCT GCTCTTCCAA TGCTGTGGAC 60 CGGGCTGGTC CTCTTGGGTC TCTTGGGATT TCCACAGACC CCAGCCCAGG GCCATGACAC 120 AGTGCAGCCC AACTTTCAAC AAGACAAGTT CCTGGGGCGC TGGTACAGCG CGGGCCTCGC 180 CTCCAATTCA AGCTGGTTCC GGGAGAAGAA AGAGCTACTG TTTATGTGCC AGACAGTGGT 240 AGCTCCCTCC ACAGAAGGCG GCCTCAACCT CACCTCTACC TTCCTAAGGA AAAACCAGTG 300 TGAGACCAAG GTGATGGTAC TGCAGCCGGC AGGGGTTCCC GGACAGTACA CCTACAACAG 360 CCCCCACTGG GGCAGCTTCC ACTCCCTCTC AGTGGTAGAA ACCGACTACG ATGAGTACGC 420 GTTCCTGTTC AGCAAGGGCA CCAAGGGCCC AGGCCAGGAC TTCCGCATGG CCACCCTCTA 480 CAGCAGAGCC CAGCTTCTGA AGGAGGAACT GAAGGAGAAA TTCATCACCT TTAGCAAGGA 540 CCAGGGCCTC ACAGAGGAGG ACATTGTTTT CCTGCCCCAA CCGGATAAGT GCATTCAAGA 600 GTAAACACAG GTGAGAGAAG TCAGTCACAG GTAACACATG GTGATGTGGC CTCAGGACTC 660 CCGTGCTCTG TCACTCTTGA GACCCAAGCC CTGGCTCCCC AAAGACCTTC TCCGCCCTCC 720 AGCTTTGCCT TGGTGGAGAA ATAAAATCCA AAGCAAGTC 759 (2) INFORMATION FOR SEQ ID NO: 5: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 876 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 5: CTCTTCCTCC TCCGGCACAC ATCGGACCTA GTAGCTGCTG AAACCATGGG CCTGGGTGTC 60 CTGTGTCTGG CCCTTGTCCT GCTTGGGGTC CTGCAGAGGC AGGCCCAGGA CTCAACTCAG 120 AACTTGATCC CTGCCCCACC TCTGATCAGT GTGCCCCTGC AGCCAGGCTT CTGGACCGAA 180 CGGTTCCAGG GCAGGTGGTT CGTTGTCGGC CTGGCAGCGA ATGCGGTCCA GAAAGAAAGA 240 CAAAGCCGCT TTACCATGTA CAGCACCATC TATGAGCTAC AGGAAGACAA TAGCTACAAC 300 GTCACTTCCA TCCTCGTCAG GGGCCAGGGC TGTCGCTACT GGATCAGAAC ATTCGTTCCA 360 AGCTCCAGGC CTGGCCAGTT CACCCTGGGG AATATTCACA GCTACCCTCA GATACAGAGC 420 TACGATGTGC AAGTGGCCGA CACTGACTAC GACCAGTTTG CCATGGTATT TTTCCAGAAG 480 ACCTCTGAAA ACAAACAGTA CTTCAAAGTC ACCCTGTACG GAAGAACCAA GGGGCTGTCC 540 GATGAACTGA AGGAGCGATT CGTCAGCTTT GCCAAGTCTC TGGGCCTCAA GGATAACAAC 600 ATCGTTTTCT CTGTTCCCAC CGACCAATGC ATTGACAACT GAACAGACGG TGAGCGTGGC 660 TGACTGGGAT GTGCAGTGGC CTGATGGTTC AGGTCCCACC TGTCTGTCTG CCGCTCCATC 720 TTTCCTGTTG CCAGAGAATC ACCTGGCTGC CCCACCAGCC ATGATTCCAT CAAGCATCTG 780 ATCCCTCTTA TTTGATCAGC TCTCCCCATC CACCTGTGTT AACGCTGCCC CACCAACGGG 840 CTCCCCCTTT CTGCTGAATA AACACATGTC CCCAAA 876 (2) INFORMATION FOR SEQ ID NO: 6: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 660 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 6: CACGAGTCCA CCCCTGCCAG GCCCAGCAGC CACCACAGCG CCTGCTTCCT CGGCCCTGAA 60 ATCATGCCCC TAGGTCTCCT GTGGCTGCCT AGCCTGTTGG GGGCTCTGCA TGCCCAGGCC 120 CAGGACTCCA CCTCAGACCT GATCCCAGCC CCACCTCTGA GCAAGGTCCC TCTGCAGCAG 180 AACTTCCAGG ACAACCAATT CCAGGGGAAG TGGTATGTGG TAGGCCTGGC AGGGAATGCA 240 ATTCTCAGAG AAGACAAAGA CCCGCAAAAG ATGTATGCCA CCATCTATGA GCTGAAAGAA 300 GACAAGAGCT ACAATGTCAC CTCCGTCCTG TTTAGGAAAA AGAAGTGTGA CTACTGGATC 360 AGGACTTTTG TTCCAGGTTG CCAGCCCGGC GAGTTCACGC TGGGCAACAT TAAGAGTTAC 420 CCTGGATTAA CGAGTTACCT CGTCCGAGTG GTGAGCACCA ACTACAACCA GCATGCTATG 480 GTGTTCTTTA AGAAAGTTTC TCAAAACAGG GAGTACTTCA AGATCACCCT CTACGGGAGA 540 ACCAAGGAGC TGACTTCGGA ACTAAAGGAG AACTTCATCC GCTTCTCCAA ATCTCTGGGC 600 CTCCCTGAAA ACCACATCGT CTTCCCTGTC CCAATCGACC AGTGTATCGA CGGCTGAGTG 660 (2) INFORMATION FOR SEQ ID NO : 7 : (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 15 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: Other Nucleic Acid (xi) SEQUENCE DESCRIPTION: SEQ ID NO : 7 : CACCATCTAT GAGCT 15 (2) INFORMATION FOR SEQ ID NO : 8 : (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 15 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: Other Nucleic Acid (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 8: CACCATCTAT GAGCT 15 (2) INFORMATION FOR SEQ ID NO: 9: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: Other Nucleic Acid (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 9: GGAGGTCGAA CTCAGA 16 (2) INFORMATION FOR SEQ ID NO: 10: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: Other Nucleic Acid (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10: CGACTGTGCA TGTCCA 16 (2) INFORMATION FOR SEQ ID NO: 11: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 10 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (n) MOLECULE TYPE: Other Nucleic Acid (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 11. GGCAATTGCC 10 (2) INFORMATION FOR SEQ ID NO: 12: (l) SEQUENCE CHARACTERISTICS- (A) LENGTH: 10 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (n) MOLECULE TYPE: Other Nucleic Acid (XI ) SEQUENCE DESCRIPTION: SEQ ID NO.12 GGCAATTACC 10 (2) INFORMATION FOR SEQ ID NO: 13. (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 10 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (n) MOLECULE TYPE: Other Nucleic Acid (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 13- GGCAATTACT 10 (2) INFORMATION FOR SEQ ID NO: 14: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 10 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (n) MOLECULE TYPE: Other Nucleic Acid (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 14: GGGAATGTCC 10

Claims: Claims We claim: 1. An isolated DNA fragment comprising a mammary gland- specific promoter, wherein the promoter promotes gene expression in a constant manner throughout the estrous cycle. 2. The isolated DNA fragment of claim 1, wherein the fragment comprises a promoter isolated from the rat NRL gene. 3. The isolated DNA fragment of claim 1 wherein the fragment comprises a promoter isolated from the NGAL gene. 4. The isolated DNA fragment of claim 3, wherein the fragment comprises the human NGAL promoter. 5. The promoter of claim 1 comprising nucleotides 1 through 2967 of SEQ ID N0:1. 6. The promoter of claim 1 comprising nucleotides 1154 through 2967 of SEQ ID NO : 1. 7. The promoter of claim 1 comprising nucleotides 1 through 2910 of SEQ ID NO: 2. 8. The promoter of claim 1 comprising nucleotides 1102 through 2910 of SEQ ID NO:2. 9. A vector comprising the promoter of claim 1. 10. A vector comprising the promoter of claim 2. 11. A vector comprising the promoter of claim 3. 12. The vector of claim 2 additionally comprising a gene sequence operably connected to the promoter. 13. The vector of claim 3 additionally comprising a gene sequence operably connected to the promoter. 14. The vector of claim 6 additionally comprising a gene sequence operably connected to the promoter. 15. The vector of claim 8 additionally comprising a gene sequence operably connected to the promoter. 16. A transgenic non-human mammal comprising the vector of claim 12. 17. A transgenic non-human mammal comprising the vector of claim 13. 18. A transgenic non-human mammal comprising the vector of claim 14. 19. A transgenic non-human mammal comprising the vector of claim 15.

Inventors: CHEN KAI-SHUN, GOULD MICHAEL N, CHEN, KAI-SHUN, GOULD, MICHAEL, N.
Applicants: WISCONSIN ALUMNI RES FOUND, WISCONSIN ALUMNI RESEARCH FOUNDATION
Application year: 1997
Main classification: C12N15
IPC codes: A01K67, A61K48, C07K14, C12N5, C12N15