JP2002512024A - Human nucleic acid sequences from fibroid tissue - Google Patents

Abstract

  (57) [Summary] A human nucleic acid sequence (mRNA, cDNA, genomic sequence) encoding a gene product or a portion thereof from uterine fibroid tissue is described and its use. Further described are the polypeptides obtained by sequence and their uses.

Description

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to a functional gene encoding a gene product or a part thereof, encoding a human nucleic acid sequence from uterine fibroid tissue, at least one biologically active polypeptide, and uses thereof. The invention further relates to the polypeptides obtained by sequence and their uses. Uterine fibroids are one of the major causes of cancer deaths in women, and new treatments are needed to combat them. Traditionally used treatments include, for example,
There are chemotherapy, hormonal treatments or surgical removal of tumor tissue, but often they do not heal completely.

[0002] Cancer phenomena frequently appear in modified cells with excessive or under-expression of a certain gene. In this case, whether the rate of degenerating expression causes malignant transformation, Or the result, which is still unclear. If such genes can be identified, it is thought that new developments in cancer treatment will be at a substantial stage. The spontaneous occurrence of cancer often precedes many mutations. These mutations can have different effects on the expression pattern in the relevant tissue, for example, under- or over-expression, and also expression of truncated genes. Some of these changes will eventually result in malignant degeneration through cascading mutations. These relationships make it difficult to solve experimentally because of the intermingling.

[0003] A data bank is used for searching for candidate genes, ie, genes that are more strongly expressed in normal tissue than in tumor tissue, and this bank consists of so-called ESTs. EST (Expression Sequence Label) is a sequence of cDNA, that is, reverse transcribed mRNA.
That is, the sequence of a molecule that reflects gene expression. The EST sequence confirms normal and degenerated cells. These databanks are offered, in part, on the market by various companies. The LifeSeq databank used here is generally between 150 and 350 nucleotides in length.

[0004] These represent a clear model for a particular gene, even when this gene is normal and very long (> 2000 nucleotides). EST identification is performed by comparing the expression patterns of normal and tumor tissues, which is important for tumor development and growth. However, the following problem occurs. That is, since the composition of the cDNA library is different, the EST sequence found may belong to a different region of the unknown gene. In this case, the relationship in which EST appears in each tissue is considered. Could be completely wrong. Only when the genes are completely known and the ESTs are sequenced to the same gene will an error be recognized.

[0005] What has now been found is that in order to reduce such errors, the entire ESTs obtained from each tissue type are assembled in advance before the expression patterns are compared with each other. Therefore, the sequence of the same gene could be integrated into the overlapping ESTs by lengthening them (see FIGS. 1, 2a and 3). By performing this lengthening and covering essentially larger gene regions in each bank, it is desirable that the above errors be further avoided. Since no software product currently exists, a program for assembling from genomic fragments was used, and by changing this, additional proprietary programs were added. A flow chart of the assembly procedure is shown in FIGS.

[0006] Now, the nucleic acid sequences of SEQ ID NO: 1 to SEQ ID NO: 31 and SEQ ID NO: 52 have been found, which play a role as candidate genes in uterine fibroids. Of particular interest are the nucleic acid sequences of SEQ ID NOs: 14-18, 30, 31, 52. Accordingly, the present invention relates to a nucleic acid sequence encoding a gene product or a part thereof, comprising: a) a nucleic acid sequence selected from the group of nucleic acid SEQ ID NOs: 14-18, 30, 31, 52; Or c) a nucleic acid sequence that is complementary to the nucleic acid sequence listed in a) or b) above.

[0007] The present invention further relates to one of the nucleic acid sequences of SEQ ID NOs: 14-18, 30, 31 or one of its complement or allelic variants and 90% to 95% complementarity of the human nucleic acid sequence. A nucleic acid sequence containing one. The present invention also relates to SEQ ID NO: 1 to SEQ ID NO: 3 whose expression in uterine fibroids is increased
1 and SEQ ID NO: 52. The invention further encompasses a portion of the nucleic acid sequences described above and thus hybridizes with SEQ ID NOs: 14-18, 30, 31, 52 when they reach a sufficient size.

[0008] The length of the nucleic acid sequence according to the invention generally represents at least 50-4500 bp,
A length of at least 150 to 4000 bp is preferred, and a length of 450 to 3500 bp is particularly excellent. The expression cassette can also be constructed according to the currently practiced method using the partial nucleic acid SEQ ID NOs: 14-18, 30, 31, 52 according to the invention, in which case at least one book on the cassette The nucleic acid sequence according to the invention can be combined with at least one control sequence or control sequences generally known to the person skilled in the art, for example a suitable promoter. The nucleic acid sequences according to the invention can be inserted in sense or antisense orientation.

[0009] Numerous expression cassettes or vectors and promoters are known in the literature and can be used. The expression cassette or vector includes: Bacterial, for example, phagescript, pBs, ΦX174, pBrewscript SK, pBsKS, pNH8a, pNH16a, pN
1. H18a, pNH46a (reticulated gene), pTrc99A, pKK223-3, pKK233-3, pDR540, pRIT5 (Pharmacia) Eukaryotic is understood, for example, pWLneo, pSV2cat, pOG44, pXT1, pSG (reticulated gene), pSVK3, pBPV, pMSG, pSVL (Pharmacia).

As control sequence or control sequence, a suitable promoter is understood.
There are two vectors that are preferred in this case, the pKK232-8 and PCM7 vectors. The following promoters can be considered individually. I.e., lacl, lacZ, T3, T7, gp
t, lambdaPR, trc, CMV, HSV thymidine kinase, SV40, LTR derived from retrovirus and mouse, metallothionein 1. The DNA sequence present in the expression cassette can encode the fusion protein, including well-known proteins and fragments of biologically active polypeptides.

[0011] Expression cassettes are also an object of the present invention. The nucleic acid fragments according to the invention can be used for the production of sufficiently long genes. The gene obtained here is also an object of the present invention. The invention also relates to the use of the nucleic acid sequence according to the invention, as well as to the gene fragments obtained using it. The nucleic acid sequence according to the invention is introduced into a host cell by means of a suitable vector, wherein the expressed genetic information obtained on the nucleic acid fragment is obtained as a non-corresponding part.

[0012] Host cells containing nucleic acid fragments are also an object of the present invention. Suitable host cells are, for example, prokaryotic cell lines, such as E. coli, or eukaryotic cell lines, such as animal or human cells or fungi. The nucleic acid sequences according to the invention can be used in sense or antisense form. Polypeptides or fragments thereof are produced by culturing host cells using current culture methods, followed by isolation and purification of the peptide or fragment, or also by today's methods. The invention further relates to nucleic acid sequences that encode at least one subsequence of a biologically active polypeptide.

The invention further relates to partial sequences of the polypeptide, so-called ORFs (open reading frames) -peptides, which are related to the sequence protocols ORF ID Nos. 32-51 and ORF
Based on ID Nos. 53-55. The invention further relates to polypeptide sequences, ie, at least one 80%
Figure 3 shows a polypeptide sequence in which homology, in particular 90% homology, has been added to the polypeptide subsequences according to the invention of ORF ID Nos. 32-51 and 53-55. The present invention also relates to antibodies, which are directed against polypeptides or fragments, encoded by a nucleic acid according to the invention having SEQ ID NO: 1-31 and SEQ ID NO: 52.

An antibody is understood in particular as a monoclonal antibody. The antibodies according to the invention are identified, inter alia, by the protein-displaying phage method. These antibodies are also an object of the present invention. The polypeptide subsequences according to the invention can be used in a protein display phage method. The polypeptide identified by this method binds to a polypeptide subsequence according to the invention, which is also an object of the invention. Similarly, the nucleic acid sequences according to the invention can be used in a protein-displaying phage method.

The polypeptides according to the invention having the sequences SEQ ID NOs: 32 to 51 and SEQ ID NOs: 53 to 55 can also be used as a tool for finding active substances against fibroids, which are also described in the book. It is an object of the invention. Similarly, an object of the present invention is to provide a polypeptide for expressing SEQ ID NOS: 1 to
It consists in using a nucleic acid sequence based on the sequence of SEQ ID NO: 31 and SEQ ID NO: 52 and can also be used as a tool for finding active substances against fibroids. The present invention has the sequences of SEQ ID NOs: 32 to 51 and SEQ ID NOs: 53 to 55,
The polypeptide subsequences found also relate to the use as a medicament for gene therapy for treating uterine fibroids or for the manufacture of a medicament for treating uterine fibroids.

The present invention also relates to a medicament comprising at least one polypeptide subsequence having the sequences SEQ ID NOs: 32-51 and SEQ ID NOs: 53-55. The nucleic acid sequences found according to the present invention may be genomic and mRNA sequences. The present invention relates to genomic genes obtained from cDNA having the sequences of SEQ ID NOs: 1 to 31, 52, their exon and intron structures and their conjugation variants, and appropriate regulatory elements such as appropriate promoters and / or enhancers. Also relates to the combined use with.

A genomic BAC-, PAC- and cosmid-library is screened using the nucleic acids according to the invention (cDNA sequences) having the sequences SEQ ID Nos: 1-31, 52 and complementary base pairs (Hybridization) a special human clone is isolated. When the BAC-, PAC- and cosmid clones isolated in this way are hybridized to intermediate chromosomes with the aid of fluorescence in situ hybridization, the corresponding chromosomal fragments are identified and the corresponding genomic Genes are generated.

[0018] BAC-, PAC- and cosmid-clones are sequenced, which reveals that the corresponding genomic genes exhibit the complete structure (promoter, enhancer, silencer, exon and intron). BAC-, PAC- and cosmid-clones can be used as unique molecules for gene transfer (see Figure 5). The invention also relates to BAC-, PAC- and cosmid-clones, which comprise:
It also corresponds to SEQ ID NOs: 1 to 31, and 52, and is used as an excipient for gene transfer with regard to the location of a functional gene and its chromosome.

Meaning of Technical Terms and Shortened Terms Nucleic Acid = Nucleic acids in the present invention are understood to be mRNAs, partial cRNAs, sufficiently long cRNAs and genomic genes (chromosomes). ORF = open reading frame, defined sequential order of amino acids derived from cRNA sequence Contig = a certain amount of DNA sequence (consensus) that can be combined into one sequence because they are so similar

Singleton = Contig Module containing only one substance = A region of a protein that represents one structural unit and has a defined sequence that appears in various proteins N = selection from nucleotides A, T, G or C X = One choice from 20 naturally occurring amino acids

Description of the alignment parameters Minimum initial match = minimum initial identification area Maximum pad / read = Maximum number of insertions Maximum mismatch% = Maximum deviation expressed in% The following examples illustrate nucleic acids according to the present invention. To explain the production of the array,
The present invention is not limited by these examples and nucleic acid sequences.

Embodiment 1 FIG . Search for candidate genes associated with tumors First, the entire EST of the corresponding tissue was extracted from the LifeSeq data bank (1
From October 997). This is next to the program GAP4 of Städenpacket
And assembled with 0% mismatch, padsperread 8 and minimum mismatch 20 as parameters. Array not input to GAP4 data bank (error)
Was assembled into a data bank with a 1% mismatch first and then a 2% mismatch again. A consensus sequence was calculated from the Contig of a databank with more than one sequence.

Singleton, a databank with only one sequence, reassembled with a new 2% mismatch due to sequences not entered in the GAP4 databank. Again, a consensus sequence was determined for Contig. 4% for all other ESTs
Newly assembled with misalignment. The consensus sequence is extracted again,
The previous consensus sequence, Singleton and the sequence not entered in the databank, were eventually assembled at a 4% mismatch. A consensus sequence was formed and, with Singleton and Error, was used as a starting basis for comparing tissues. Performing this procedure ensured that the entire sequence of unrelated gene regions was represented under the parameters used.

FIGS. 2 b 1-2 b 4 illustrate the elongation of uterine fibroid tissue EST. In this way, the assembled sequences for each tissue were subsequently compared with each other using the same program (FIG. 3). For this purpose, the entire first cell sequence was first entered into the data bank. (Thus, it was important that they were independent of each other.)

Next, all sequences of the second cell were all compared to the first cell. The resulting sequence is unique for the first or second tissue and both appear. For the second cell, the frequency ratio was evaluated for each tissue. The entire program evaluation involving the assembled sequence was developed by itself.

Further investigations were performed on all sequences that occurred more than four times in any one of the compared tissues, and all sequences that appeared very often at least five times in one of both tissues. Was. These arrays were used in the Electronics Northern method (Example
2.1), the overall distribution of tumor cells and normal cells was examined (FIGS. 4a and 4b).
reference). Relevant candidate cells were then extended using the whole Incyte EST and all published data bank ESTs (see FIG. 3). Subsequently, the sequence and its possible translation into proteins were compared with all nucleotide and protein databanks, and regions which could also be encoded as proteins were examined.

Embodiment 2 FIG . An algorithm for identification and extension of a partial sequence of cDNA by a changeable expression pattern. Next, an algorithm for finding a gene whose expression is excessive or underexpressed will be described. The individual steps are also summarized in the flow diagram to make the whole picture better visible (see FIG. 4b).

2.1 Electronics Northern Blotting DNA subsequences S, eg, for individual ESTs or EST Contigs, are defined by homologous sequences in a (personal or public) EST library organized according to various tissues. However, this is a standard program for homolog search, such as BLAST (Altschul, SF, GishW., Miller, W., Myers, EWund Lipma).
n, DJ (1990) J. Mol. Biol., 215, 403-410), BLAST2 (Altschul, SF, Ma
dden, TL, Sch-ffer, AA, Zhang, J., Zhang, Z., Miller, W., und Lipma
n, DJ (1997) Nucleic Acids Research 25, 3389-3402) or FASTA (Pearso
n, WR, und Lipman, DJ (1988) Proc. Natl. Acad. Sci. USA 85 2444-2448). The appearance frequency (relative value or absolute value) specific to the tissue of the DNA partial sequence S confirmed by this is expressed as an electronic northern blot method.

2.1.1 SEQ ID NO: 30 was found, similar to the method described in 2.1, but appears more frequently in fibroid tissue than in normal tissue. The results are shown below.

[Table 1] 2.1.2 SEQ ID NO: 31 was found, similar to the method described in 2.1, but appears more frequently in fibroid tissue than in normal tissue. The results are shown below.

[0030]

[Table 2]

[0031]

[Table 3]

[0032]

[Table 4]

[0033]

[Table 5]

[0034]

[Table 6]

[0035]

[Table 7]

[0036]

[Table 8]

[0037]

[Table 9]

[0038]

[Table 10]

[0039]

[Table 11]

[0040]

[Table 12]

[0041]

[Table 13]

[0042]

[Table 14]

[0043]

[Table 15]

[0044]

[Table 16]

[0045]

[Table 17]

[0046]

[Table 18]

[0047]

[Table 19]

[0048]

[Table 20]

[0049]

[Table 21]

[0050]

[Table 22]

[0051]

[Table 23]

[0052]

[Table 24]

[0053]

[Table 25]

[0054]

[Table 26]

[0055]

[Table 27]

[0056]

[Table 28]

[0057]

[Table 29]

[0058]

[Table 30]

[0059]

[Table 31]

[0060]

[Table 32]

[0061]

[Table 33]

[0062]

[Table 34]

2.2 Fisher Test To determine if the partial sequence S of a gene appears significantly more frequently or rarely in a library against normal tissues than in a library against mutant tissues, Fisher's exact test, Standard method (Hays, W.
L., (1991) Statistics, Harcourt Brace College Publishers, Fort Worth). According to the zero hypothesis, both libraries are indistinguishable in frequency for sequences homologous to S. If the zero hypothesis can be rejected with sufficiently high certainty, the gene associated with S is taken as an interesting candidate for the oncogene and the next step is to try to achieve extension of this sequence. Example 3 Automatic extension of partial sequence Automatic extension of partial sequence S is performed in the following three stages. 1. All homologous sequences are determined as S using BLAST from the total amount of available sequences. 2. These sequences were compiled into the standard program GAP4 (Bonfield, JK, Smith, KF, Stade
n Assemble using R. (1995), Nucleic Acids Research 23 4992-4999) (contig generation). 3. The consensus sequence C is calculated based on the assembled sequence. The consensus sequence C will generally be longer than the starting sequence S. Thus, the electronic Northern blot will be different from that of the Northern blot for S.
Again, Fisher's test determines whether the alternative hypothesis of the difference for equal expression in both libraries can be maintained. If so, try to extend C in the same manner as S. In this iteration, the alternative hypothesis is denied (if Ho Exit: interruption criterion I) or automatic extension becomes impossible (while Cj> Cj-1: interruption criterion I)
Continue with the consensus sequence Cj (j: index of repetition) obtained in each case until I). In the case of interruption criterion II, the consensus sequence present after the last repeat gives a complete or near-complete sequence of the gene that can be associated with cancer with high statistical accuracy. By analogy with the above example, the nucleic acid sequences listed in Table I could be found from fibroid tissue. Furthermore, for each nucleic acid sequence, the peptide sequences (ORF) summarized in Table II could be defined. In this case, a small number of nucleic acid sequences cannot be associated with a peptide, and some nucleic acid sequences can be associated with one or more peptides. As mentioned above, both the determined nucleic acid sequence and the peptide sequence associated with the nucleic acid sequence are an object of the present invention. Example 4. Mapping of the Nucleic Acid Sequence on the Human Genome The mapping of the human genome was performed using the Stanford G3 hybrid panel (Stewart et al., 1) sold by Research Genetics (Huntsville, Ala.).
997). This panel consists of 83 genomic DNAs of the human hamster hybrid cell line, enabling a resolution of 500 kilobases. The hybrid cell line was obtained by fusion of irradiated human diploid cells with Chinese hamsters. Retention patterns of human chromosome segments were defined using gene-specific primers in the polymerase chain reaction and analyzed using software provided by the Stanford RH server (http://www.Stanford.edu/RH/rhs).
erver # form2.html). This program uses the S
Define TS marker. Corresponding cytogenetic linkages can be found in the Genome Database (GDB
) Using the map view program (http: //gdbwww.dkfz-heidelberg/de). Along with the mapping of genes on the human chromosome set by various experimental methods, it is possible to define the location of the genes there by bioinformatic methods. For this purpose, the known program e-PCR was used (SchulerGD (
1998) Electronic PCR: bridging The gap between genome mapping and genome s
equencing.Trends Biotechnol 16: 456-459, Schuler GD (1997) .Sequence mappin
g by electronic PCR. Genome Res 7: 541-550). The data bank used here is no longer the data bank listed in the literature, but a public data bank RHdb (http: //
www.ebi.ac.uk/RHdb/ndex.html). By analogy with the hybrid panel mapping, the above software and the software of the Whitehead Laboratory (http://carbon.wi.mit.edu:8000/cgi-bin/contig/rhmapper.p
The result was evaluated in l).

[0064]

[Table 35]

[0065]

[Table 36]

[0066]

[Table 37]

[0067]

[Sequence list]

 (1) General information: (ii) Title of the invention: human nucleic acid sequence from myometrial tissue (iii) Number of sequences: 55 (2) Information of SEQ ID NO: 1 (i) Sequence characteristics: (A) Length Length: 779 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: linear (ii) Molecular type: partial cDNA generated from single-stranded EST by assembly and editing (Iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) organ: (vii) direct origin: (A) library: cDNA library (xi) Description of sequence: SEQ ID NO: 1

[0068]

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(2) Information of SEQ ID NO: 2: (i) Sequence characteristics: (A) Length: 2310 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded ESTs by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Sequence description: SEQ ID NO: 2

[0070]

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(2) Information of SEQ ID NO: 3: (i) Sequence characteristics: (A) Length: 854 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded ESTs by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Description of sequence: SEQ ID NO: 3

[0072]

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(2) Information of SEQ ID NO: 4: (i) Sequence characteristics: (A) Length: 1112 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded ESTs by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Description of sequence: SEQ ID NO: 4

[0074]

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(2) Information of SEQ ID NO: 5: (i) Sequence characteristics: (A) Length: 1051 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded ESTs by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Description of sequence: SEQ ID NO: 5

[0076]

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(2) Information of SEQ ID NO: 6: (i) Sequence characteristics: (A) Length: 1516 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded ESTs by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Description of sequence: SEQ ID NO: 6

[0078]

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(2) Information of SEQ ID NO: 7: (i) Sequence characteristics: (A) Length: 2367 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded ESTs by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Description of sequence: SEQ ID NO: 7

[0080]

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(2) Information of SEQ ID NO: 8: (i) Sequence characteristics: (A) Length: 568 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded ESTs by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Description of sequence: SEQ ID NO: 8

[0082]

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(2) Information of SEQ ID NO: 9: (i) Sequence characteristics: (A) Length: 1775 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded ESTs by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Description of sequence: SEQ ID NO: 9

[0084]

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(2) Information of SEQ ID NO: 10: (i) Sequence characteristics: (A) Length: 509 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded ESTs by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Description of sequence: SEQ ID NO: 10

[0086]

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(2) Information of SEQ ID NO: 11: (i) Sequence characteristics: (A) Length: 2191 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded ESTs by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Description of sequence: SEQ ID NO: 11

[0088]

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(2) Information of SEQ ID NO: 12: (i) Sequence characteristics: (A) Length: 1769 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded ESTs by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Description of sequence: SEQ ID NO: 12

[0090]

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(2) Information of SEQ ID NO: 13: (i) Sequence characteristics: (A) Length: 1026 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded ESTs by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Description of sequence: SEQ ID NO: 13

[0092]

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(2) Information of SEQ ID NO: 14: (i) Sequence characteristics: (A) Length: 676 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded ESTs by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Description of sequence: SEQ ID NO: 14

[0094]

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(2) Information of SEQ ID NO: 15: (i) Sequence characteristics: (A) Length: 1254 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded ESTs by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Description of sequence: SEQ ID NO: 15

[0096]

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(2) Information of SEQ ID NO: 16: (i) Sequence characteristics: (A) Length: 537 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded ESTs by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Description of sequence: SEQ ID NO: 16

[0098]

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(2) Information of SEQ ID NO: 17: (i) Sequence characteristics: (A) Length: 823 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded ESTs by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Description of sequence: SEQ ID NO: 17

[0100]

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(2) Information of SEQ ID NO: 18: (i) Sequence characteristics: (A) Length: 1082 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded ESTs by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Description of sequence: SEQ ID NO: 18

[0102]

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(2) Information of SEQ ID NO: 19: (i) Sequence characteristics: (A) Length: 1548 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded ESTs by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Sequence description: SEQ ID NO: 19

[0104]

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(2) Information of SEQ ID NO: 20: (i) Sequence characteristics: (A) Length: 844 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded ESTs by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Description of sequence: SEQ ID NO: 20

[0106]

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(2) Information of SEQ ID NO: 21: (i) Sequence characteristics: (A) Length: 862 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded ESTs by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Description of sequence: SEQ ID NO: 21

[0108]

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(2) Information of SEQ ID NO: 22: (i) Sequence characteristics: (A) Length: 546 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded ESTs by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Description of sequence: SEQ ID NO: 22

[0110]

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(2) Information of SEQ ID NO: 23: (i) Sequence characteristics: (A) Length: 1591 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded ESTs by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Description of sequence: SEQ ID NO: 23

[0112]

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(2) Information of SEQ ID NO: 24: (i) Sequence characteristics: (A) Length: 441 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded ESTs by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Description of sequence: SEQ ID NO: 24

[0114]

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(2) Information of SEQ ID NO: 25: (i) Sequence characteristics: (A) Length: 1131 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded ESTs by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Description of sequence: SEQ ID NO: 25

[0116]

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(2) Information of SEQ ID NO: 26: (i) Sequence characteristics: (A) Length: 1071 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded ESTs by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Description of sequence: SEQ ID NO: 26

[0118]

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(2) Information of SEQ ID NO: 27: (i) Sequence characteristics: (A) Length: 896 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded ESTs by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Description of sequence: SEQ ID NO: 27

[0120]

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(2) Information of SEQ ID NO: 28: (i) Sequence characteristics: (A) Length: 1050 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded ESTs by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Description of sequence: SEQ ID NO: 28

[0122]

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(2) Information of SEQ ID NO: 29: (i) Sequence characteristics: (A) Length: 581 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded ESTs by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Description of sequence: SEQ ID NO: 29

[0124]

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(2) Information of SEQ ID NO: 30: (i) Sequence characteristics: (A) Length: 264 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded ESTs by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Description of sequence: SEQ ID NO: 30

[0126]

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(2) Information of SEQ ID NO: 31: (i) Sequence characteristics: (A) Length: 111 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded ESTs by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Description of sequence: SEQ ID NO: 31

[0128]

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(2) Information of SEQ ID NO: 32: (i) Sequence characteristics: (A) Length: 76 amino acids (B) Type: protein (C) Number of chains: single chain (D) Topology: straight Linear (ii) molecular type: ORF (iii) hypothetical: yes (vi) origin (A) organism: human (xi) description of sequence: SEQ ID NO: 32

[0130]

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(2) Information of SEQ ID NO: 33: (i) Sequence characteristics: (A) Length: 72 amino acids (B) Type: protein (C) Number of chains: single chain (D) Topology: straight Linear (ii) molecular type: ORF (iii) hypothetical: yes (vi) origin (A) organism: human (xi) description of sequence: SEQ ID NO: 33

[0132]

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(2) Information of SEQ ID NO: 34: (i) Sequence characteristics: (A) Length: 70 amino acids (B) Type: protein (C) Number of chains: single chain (D) Topology: direct Linear (ii) molecular type: ORF (iii) hypothetical: yes (vi) origin (A) organism: human (xi) description of sequence: SEQ ID NO: 34

[0134]

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(2) Information of SEQ ID NO: 35: (i) Sequence characteristics: (A) Length: 60 amino acids (B) Type: protein (C) Number of chains: single chain (D) Topology: straight Linear (ii) molecular type: ORF (iii) hypothetical: yes (vi) origin (A) organism: human (xi) description of sequence: SEQ ID NO: 35

[0136]

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(2) Information of SEQ ID NO: 36: (i) Sequence characteristics: (A) Length: 63 amino acids (B) Type: protein (C) Number of chains: single chain (D) Topology: straight Linear (ii) molecular type: ORF (iii) hypothetical: yes (vi) origin (A) organism: human (xi) description of sequence: SEQ ID NO: 36

[0138]

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(2) Information of SEQ ID NO: 37: (i) Sequence characteristics: (A) Length: 170 amino acids (B) Type: protein (C) Number of chains: single chain (D) Topology: direct Linear (ii) molecular type: ORF (iii) hypothetical: yes (vi) origin (A) organism: human (xi) description of sequence: SEQ ID NO: 37

[0140]

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(2) Information of SEQ ID NO: 38: (i) Sequence characteristics: (A) Length: 144 amino acids (B) Type: protein (C) Number of chains: single chain (D) Topology: direct Linear (ii) molecular type: ORF (iii) hypothetical: yes (vi) origin (A) organism: human (xi) description of sequence: SEQ ID NO: 38

[0142]

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(2) Information of SEQ ID NO: 39: (i) Sequence characteristics: (A) Length: 178 amino acids (B) Type: protein (C) Number of chains: single chain (D) Topology: direct Linear (ii) molecular type: ORF (iii) hypothetical: yes (vi) origin (A) organism: human (xi) description of sequence: SEQ ID NO: 39

[0144]

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(2) Information of SEQ ID NO: 40: (i) Sequence characteristics: (A) Length: 89 amino acids (B) Type: protein (C) Number of chains: single chain (D) Topology: direct Linear (ii) molecular type: ORF (iii) hypothetical: yes (vi) origin (A) organism: human (xi) description of sequence: SEQ ID NO: 40

[0146]

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(2) Information of SEQ ID NO: 41: (i) Sequence characteristics: (A) Length: 95 amino acids (B) Type: protein (C) Number of chains: single chain (D) Topology: direct Linear (ii) molecular type: ORF (iii) hypothetical: yes (vi) origin (A) organism: human (xi) description of sequence: SEQ ID NO: 41

[0148]

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(2) Information of SEQ ID NO: 42: (i) Sequence characteristics: (A) Length: 154 amino acids (B) Type: protein (C) Number of chains: single chain (D) Topology: direct Linear (ii) molecular type: ORF (iii) hypothetical: yes (vi) origin (A) organism: human (xi) description of sequence: SEQ ID NO: 42

[0150]

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(2) Information of SEQ ID NO: 43: (i) Sequence characteristics: (A) Length: 79 amino acids (B) Type: protein (C) Number of chains: single chain (D) Topology: direct Linear (ii) molecular type: ORF (iii) hypothetical: yes (vi) origin (A) organism: human (xi) description of sequence: SEQ ID NO: 43

[0152]

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(2) Information of SEQ ID NO: 44: (i) Sequence characteristics: (A) Length: 82 amino acids (B) Type: protein (C) Number of chains: single chain (D) Topology: direct Linear (ii) molecular form: ORF (iii) hypothetical: yes (vi) origin (A) organism: human (xi) description of sequence: SEQ ID NO: 44

[0154]

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(2) Information of SEQ ID NO: 45: (i) Sequence characteristics: (A) Length: 68 amino acids (B) Type: protein (C) Number of chains: single chain (D) Topology: straight Linear (ii) molecular type: ORF (iii) hypothetical: yes (vi) origin (A) organism: human (xi) description of sequence: SEQ ID NO: 45

[0156]

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(2) Information of SEQ ID NO: 46: (i) Sequence characteristics: (A) Length: 87 amino acids (B) Type: protein (C) Number of chains: single chain (D) Topology: direct Linear (ii) molecular type: ORF (iii) hypothetical: yes (vi) origin (A) organism: human (xi) description of sequence: SEQ ID NO: 46

[0158]

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(2) Information of SEQ ID NO: 47: (i) Sequence characteristics: (A) Length: 51 amino acids (B) Type: protein (C) Number of chains: single chain (D) Topology: straight Linear (ii) molecular type: ORF (iii) hypothetical: yes (vi) origin (A) organism: human (xi) description of sequence: SEQ ID NO: 47

[0160]

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(2) Information of SEQ ID NO: 48: (i) Sequence characteristics: (A) Length: 20 amino acids (B) Type: protein (C) Number of chains: single chain (D) Topology: straight Linear (ii) molecular type: ORF (iii) hypothetical: yes (vi) origin (A) organism: human (xi) description of sequence: SEQ ID NO: 48

[0162]

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(2) Information of SEQ ID NO: 49: (i) Sequence characteristics: (A) Length: 36 amino acids (B) Type: protein (C) Number of chains: single chain (D) Topology: straight Linear (ii) molecular type: ORF (iii) hypothetical: yes (vi) origin (A) organism: human (xi) description of sequence: SEQ ID NO: 49

[0164]

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(2) Information of SEQ ID NO: 50: (i) Sequence characteristics: (A) Length: 26 amino acids (B) Type: protein (C) Number of chains: single chain (D) Topology: direct Linear (ii) molecular type: ORF (iii) hypothetical: yes (vi) origin (A) organism: human (xi) description of sequence: SEQ ID NO: 50

[0166]

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(2) Information of SEQ ID NO: 51: (i) Sequence characteristics: (A) Length: 25 amino acids (B) Type: protein (C) Number of chains: single chain (D) Topology: straight Linear (ii) molecular type: ORF (iii) hypothetical: yes (vi) origin (A) organism: human (xi) description of sequence: SEQ ID NO: 51

[0168]

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(2) Information of SEQ ID NO: 52: (i) Sequence characteristics: (A) Length: 3665 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded EST by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Description of sequence: SEQ ID NO: 52

[0170]

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[0171]

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(2) Information of SEQ ID NO: 53: (i) Sequence characteristics: (A) Length: 301 base pairs (B) Type: nucleic acid (C) Number of strands: single strand (D) Topology: Linear (ii) molecular form: partial cDNA generated from single-stranded EST by assembly and editing (iii) hypothetical: NO (iii) antisense: NO (vi) origin: (A) organism: human (C) Organ: (vii) Direct origin: (A) Library: cDNA library (xi) Description of sequence: SEQ ID NO: 53

[0173]

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(2) Information of SEQ ID NO: 54: (i) Sequence characteristics: (A) Length: 112 amino acids (B) Type: protein (C) Number of chains: single chain (D) Topology: direct Linear (ii) molecular type: ORF (iii) hypothetical: yes (vi) origin (A) organism: human (xi) description of sequence: SEQ ID NO: 54

[0175]

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(2) Information of SEQ ID NO: 55: (i) Sequence characteristics: (A) Length: 107 amino acids (B) Type: protein (C) Number of chains: single chain (D) Topology: direct Linear (ii) molecular type: ORF (iii) hypothetical: yes (vi) origin (A) organism: human (xi) description of sequence: SEQ ID NO: 55

[0177]

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[Sequence list]

[Brief description of the drawings]

FIG. 1 shows a systematic gene search in the Incyte Life Seq databank.

FIG. 2a illustrates the principle of the EST assembly.

FIG. 2b1 shows the overall principle of the EST assembly.

FIG. 2b2 shows the overall principle of the EST assembly.

FIG. 2b3 shows the overall principle of the EST assembly.

FIG. 2b4 shows the overall principle of the EST assembly.

FIG. 3 shows silico subtraction of gene expression in various tissues.

FIG. 4a shows the measurement of tissue-specific expression by the electronic Northern method.

FIG. 4b shows the electronic Northern method.

FIG. 5 shows the isolation of genomic BAC-, PAC-clones.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) C12N 5/10 C12P 21/08 C12P 21/02 (C12N 1/21 21/08 C12R 1:19) // (C12N 1/21 C12N 15/00 ZNAA C12R 1:19) 5/00 B (72) Inventor Schmidt, Almin Germany 14-14 Berlin, Laubacher Strasse 6 / tubite (72) Inventor Pilarskiy, Christian Germany, Day-01474 Schoenfeld-Bishik, Heinrich-Lange-Strasse 13 Tse (72) Inventor Dar, Edgar Germany, Day-14480 Potsdam, Eleonore-Progeska-Strasse 6 (72) Inventor Rosenta Le, Andre Germany, Day 10115 Berlin, Coppenplatz 10F term (reference) 4B024 AA01 AA12 BA80 CA04 CA07 DA02 DA03 DA06 DA12 EA04 FA02 FA06 GA11 HA01 HA11 HA17 4B064 AG01 AG27 CA02 CA06 CA10 CA19 CA20 CC24 DA05 DA14 4B065 AA26X AA72X AA93X AA93Y AB01 BA02 CA24 CA25 CA44 CA46 4H045 AA10 AA11 AA20 AA30 BA10 BA18 BA19 BA20 BA21 CA41 DA76 EA28 EA51 FA72 FA74

Claims (38)

[Claims] 1. A gene product or a part thereof, which encodes: a) a nucleic acid sequence selected from the group of SEQ ID NOs: 14 to 18, 30, 31, and 52; b) a nucleic acid sequence listed in a) above. An allelic variant, or c) a nucleic acid sequence that shows complementarity to the nucleic acid sequence listed under a) or b) above. 2. A nucleic acid sequence based on one of SEQ ID NOs: 14-18, 30, 31, 52 or a complement or allelic variant thereof. 3. Nucleic acid SEQ ID NO: 1 to SEQ ID NO: 31 and SEQ ID NO: 52, wherein the expression is increased in uterine fibroid tissue. 4. Including a functional gene and its chromosome localization, SEQ ID NO: 1
BAC, PAC and cosmid-clones corresponding to SEQ ID NO: 31 and SEQ ID NO: 52 and used as vehicles for gene transfer. 5. The nucleic acid sequence according to claim 1, wherein the nucleic acid sequence is 90% homologous to the human nucleic acid sequence. 6. The nucleic acid sequence according to claim 1, wherein the nucleic acid sequence is 95% homologous to the human nucleic acid sequence. 7. The method according to claim 1, which comprises a part of the nucleic acid sequence according to claim 1.
A nucleic acid sequence having a size sufficient to form a hybrid with the sequence described in any one of claims 6 to 6. 8. The nucleic acid sequence according to claim 1, wherein the fragment has a length of at least 50 to 4500 bp. 9. The nucleic acid sequence according to claim 1, wherein the fragment has a length of at least 50 to 4000 bp. 10. The nucleic acid sequence according to claim 1, which encodes at least one partial sequence of a biologically active polypeptide. 11. An expression cassette comprising a nucleic acid fragment or a sequence according to any one of claims 1 to 9 together with at least one control sequence or control sequence. 12. An expression cassette comprising a nucleic acid fragment or the sequence according to claim 11, wherein the control sequence or control sequence is a suitable promoter. 13. The DNA sequence present in the cassette encodes a fusion protein, wherein the fusion protein includes well-known proteins and biologically active polypeptide fragments. 13. The expression cassette according to any one of 11 and 12. 14. Use of the nucleic acid sequence according to claims 1 to 10 for producing a gene that is sufficiently long. 15. A DN comprising the gene obtained by use of claim 14.
A fragment. 16. The heterologous part of gene information that can be expressed,
A host cell containing the nucleic acid fragment according to any one of claims 10 to 10. 17. The host cell according to claim 16, wherein the system is a prokaryotic or eukaryotic cell line. 18. The method according to claim 16, wherein the prokaryotic cell line is Escherichia coli and the eukaryotic cell line is an animal, human or yeast cell line. Host cells. 19. A method for producing a polypeptide or fragment, wherein the host cell according to claim 16 is cultured. 20. An antibody against a polypeptide or fragment encoded by the nucleic acids of SEQ ID NOs: 1-31 and 52. 21. The antibody according to claim 20, which is a monoclonal antibody. 22. The antibody according to claim 20, which is a protein-displaying phage antibody. 23. A polypeptide partial sequence according to SEQ ID NOs: 32 to 51 and SEQ ID NOs: 53 to 55. 24. The polypeptide subsequence of claim 23, which has at least 80% homology with the polypeptide sequence. 25. A polypeptide that emerges from a protein-displaying phage and is capable of binding to the polypeptide subsequence of claim 23. 26. The polypeptide subsequence of claim 23, which has at least 90% homology with the polypeptide sequence. 27. A tool for finding active substances against fibroids,
Use of a polypeptide partial sequence set forth in SEQ ID NOs: 32 to 51 and SEQ ID NOs: 53 to 55. 28. Use of a nucleic acid sequence according to SEQ ID NOs: 1-31 and 52 for expressing a polypeptide which can be used as a tool for finding active substances against uterine fibroids. 29. Nucleic acid SEQ ID NOS: 1-31 in sense or antisense form
And use of SEQ ID NO: 52. 30. Use of the polypeptide partial SEQ ID NO: 32 to SEQ ID NO: 51 and SEQ ID NO: 53 to 55 as a medicament in gene therapy for treating uterine fibroids. 31. Use of a polypeptide partial SEQ ID NO: 32 to SEQ ID NO: 51 and SEQ ID NO: 53 to 55 for the manufacture of a medicament for treating uterine fibroids. 32. A medicament comprising at least one polypeptide subsequence of SEQ ID NO: 32 to SEQ ID NO: 51 and SEQ ID NO: 53 to 55. 33. The nucleic acid sequence according to claim 1, which is a genomic sequence. 34. The nucleic acid sequence according to claim 1, which is an mRNA sequence. 35. A genomic gene obtained from cDNA having SEQ ID NOS: 1-31 and SEQ ID NO: 52, its promoter, enhancer, silencer, exon structure, intron structure and its junctional variant. 36. Use of the genomic gene according to claim 33 in combination with a suitable regulatory element. 37. Use according to claim 36, wherein said regulatory element is a suitable promoter and / or enhancer. 38. The nucleic acid sequence according to claim 1, wherein the fragment has a length of at least 300 to 3500 bp.