RU2793635C2 - Preoperative risk stratification based on pde4d7 and dhx9 expression - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: group of inventions relates to preoperative risk stratification in a subject suffering from prostate cancer. A method is proposed, including determining the expression profile of the PDE4D7 gene and the expression profile of the DHX9 gene in a biological sample obtained from a subject. Based on these profiles, a preoperative predictive risk score is determined. In this case, the expression profiles of the PDE4D7 and DHX9 genes are combined with a regression function that was obtained on the basis of a population of subjects suffering from prostate cancer. The risk score indicates the risk of biochemical recurrence (BCR), clinical recurrence (CR), or prostate cancer-specific mortality. Also proposed are a diagnostic kit and its use in this method, the use of a gene expression profile for PDE4D7 and a gene expression profile for DHX9 in this method, as well as an appropriate machine-readable carrier.

EFFECT: inventions allow to provide improved stratification of the subject in the preoperative condition, which can lead to more correct decisions on primary treatment.

12 cl, 6 dwg, 29 tbl

Description

FIELD OF TECHNOLOGY

The present invention relates to a method for preoperative risk stratification in a subject suffering from prostate cancer. In addition, the present invention relates to a diagnostic kit, to the use of said diagnostic kit in a method for preoperative risk stratification in a subject suffering from prostate cancer, to the use of a gene expression profile for phosphodiesterase 4D variant 7 (PDE4D7) and a gene expression profile for DExH-box- helicase 9 (DHX9) in preoperative risk stratification in a subject suffering from prostate cancer, as well as to an appropriate computer program product.

BACKGROUND OF THE INVENTION

Cancer is a class of diseases in which a group of cells exhibit uncontrolled reproduction, invasion, and sometimes metastasis. These three malignant properties of different types of cancer distinguish them from benign tumors, which are self-limiting, incapable of invasion or metastasis. Prostate cancer (PCa) is the most common non-skin malignant neoplasm in men, with about 1.1 million new cases diagnosed worldwide in 2012 (see Ferlay J. et al., GLOBOCAN 2012 v1.0, Cancer Incidence and Mortality Worldwide: IARC CancerBase No. 11 [Internet], Lyon, France, International Agency for Research on Cancer, 2013).

Due to population aging, the incidence of prostate cancer will continue to increase in the coming years (see Quon H. et al., "Dramatic increase in prostate cancer cases by 2021", BJU International, Vol. 108, No. 11, pages 1734-1738 , 2011). Routine diagnosis with blood levels of prostate-specific antigen (PSA), digital rectal examination (DRE), and transrectal ultrasound (TRUS) leads to significant primary overdiagnosis of non-cancerous benign conditions of the prostate (see Bangma C.H. et al., “Overdiagnosis and overtreatment of early detected prostate cancer", World Journal of Urology, Vol. 25, No. 1, pages 3-9, 2007, and Schröder F.H. et al., "Screening and Prostate-Cancer Mortality in a Randomized European Study", The New England Journal of Medicine Vol. patients have significant complications (urosepsis, bleeding, urinary retention) and cost more than US$2 billion in total (~US$2100 per biopsy procedure). At least 4 in 100 biopsy-negative men are likely to be hospitalized due to side effects, and 9 in 10,000 biopsied patients are at risk of death due to the current procedure (see Nam R.K. et al. , "Increasing hospital admission rates for urological complications after transrectal ultrasound guided biopsy", Journal of Urology, Vol. 183, No. 3, pages 963-968, 2010).

In the US, of the approximately 250,000 new cases of prostate cancer diagnosed per year, approximately 200,000 are initially characterized as localized disease (see Snyder C.F. et al., "How does initial treatment choice affect short-term and long-term costs for clinically localized prostate cancer?", Cancer, Vol. 116, No. 23, pages 5391-5399, 2010, and Cooperberg M. R. et al., "Contemporary trends in low risk prostate cancer: risk assessment and treatment", Journal of Urology, Vol. 173, No. 3 , Pt. 2, pages 14-19, 2007), that is, as a cancer limited to the organ of the prostate gland. This condition is curable to some extent with primary treatment approaches such as radiation therapy or, in particular, partial or complete removal of the prostate gland through surgery (prostatectomy). However, these interventions are usually accompanied by serious side effects, in particular urinary incontinence and/or erectile dysfunction, as very common consequences of prostatectomy. Up to 50% of men undergoing radical prostatectomy suffer from urinary incontinence. Studies have shown that 15% to 50% of men still report such problems a year after surgery. Erection problems are also serious side effects of radical prostatectomy (RP). Only about half of the operated men can partially restore their ability to have erections. In addition, all routinely used treatments for localized PCa are expensive (typically in the order of US$20-30,000) and in the US result in a total direct cost of US$5 billion per year.

Among the ~200,000 men in the United States with clinically localized disease at diagnosis, up to 50% are at very low or low risk of developing cancer (see Bangma C.H. and Roobol M.J., "Defining and predicting indolent and low prostate cancer", Critical Reviews in Oncology /Hematology, Vol.83, No.2, pages 235-241, 2012). Accordingly, the US National Comprehensive Cancer Network (NCCN) recently revised its guidelines for the treatment of PCa to expand active surveillance (AS) as a gentle and convenient treatment alternative for patients with low-risk disease (see www. nccn.org). The quality of life of such patients is significantly improved when eligible patients are referred to AS, compared with primary-treated men, and the 5-year AS cost is reported to be less than US$10,000 per patient (see Snyder C.F. et al.).

Moreover, if surgery is chosen as the preferred treatment for a particular patient (versus AS), stratification of the extent of surgery according to the potential aggressiveness of the patient's tumor provides a significant advantage. For example, nerve-sparing techniques may be applied more widely to men with low-risk predictive disease to minimize potency-related adverse effects of radical prostatectomy. Similarly, according to the most recent European Association of Urology (EAU) guidelines for prostate cancer, extensive lymph node dissection is recommended when a high risk of cancer is predicted, although the procedure is complex, time-consuming and associated with a higher incidence. complications compared to more limited procedures (see Heidenreich A. et al., Guidelines on Prostate Cancer, European Association of Urology, 2012). Therefore, while less limited lymph node dissection has been shown to miss approximately 50% of lymph node metastases (see Heidenreich A. et al., "Extended pelvic lymphadenectomy in patients undergoing radical prostatectomy: high incidence of lymph node metastasis ", Journal of Urology, Vol. 167, No. 4, pages 1681-1686, 2002, and Bader P. et al, "Is a limited lymph node dissection and adequate staging procedure for prostate cancer?", Journal of Urology, Vo 168, No. 2, pages 514-518, 2002), management of men with localized prostate cancer requires highly accurate preoperative predictions of the potential aggressiveness of an individual tumor in order to provide the most optimal care for each patient.

WO 2014/028884 A2 discloses methods, systems and kits for diagnosing, predicting and determining the progression of cancer in a subject. Also disclosed are methods, systems, and kits for determining a method of treating cancer in a subject. Methods, systems and kits include the analysis of biomarkers based on expression. In addition, probe sets are disclosed for use in assessing a subject's cancer status.

BRIEF DESCRIPTION OF THE INVENTION

The purpose of the present invention is to provide a method for preoperative risk stratification in a subject suffering from prostate cancer, which can provide more accurate preoperative predictions of the potential aggressiveness of an individual tumor. Another object of the present invention is to provide a diagnostic kit, the use of said diagnostic kit in a method for preoperative risk stratification in a subject suffering from prostate cancer, the use of a gene expression profile for phosphodiesterase 4D variant 7 (PDE4D7) and a gene expression profile for DExH-box helicase 9 (DHX9) in a preoperative risk stratification in a subject with prostate cancer, as well as the corresponding computer software product.

According to a first aspect of the present invention, there is provided a method for preoperative risk stratification in a subject suffering from prostate cancer, comprising:

- determining the expression profile of the gene for variant 7 of phosphodiesterase 4D (PDE4D7) in a biological sample obtained from the subject,

- determining the expression profile of the gene for DExH-box helicase 9 (DHX9) in the same or different biological sample obtained from the subject, and

- determining a preoperative predictive risk score for the subject based on the gene expression profile for PDE4D7 and the gene expression profile for DHX9.

It is known that the cAMP signaling pathway plays an important role both in the development and progression of prostate cancer (see Merkle D. and Hoffmann R., “Roles of cAMP and cAMP-dependent protein kinase in the progression of prostate cancer: Cross- talk with the androgen receptor", Cellular Signaling, Vol. 23, No. 3, pages 507-515, 2011). While the adenylyl cyclase family is responsible for cAMP synthesis, cyclic nucleotide phosphodiesterases (PDEs) appear to be the only cellular mechanism for its degradation. PDEs provide signal termination and, importantly, compartmentalization of cAMP signaling in the three-dimensional matrix of cells. This is achieved through spatially discrete degradation of cAMP by subpopulations of various PDE isoforms sequestered by localized anchor proteins/signalosomes (see, e.g., Conti M. and Beavo J., “Biochemistry and physiology of cyclic nucleotide phosphodiesterases: essential components in cyclic nucleotide signaling", Annual Review of Biochemistry, Vol. 76, pages 481-511, 2007). Thus, changes in the expression and/or activity of various PDE isoforms may alter downstream signaling pathways during disease development and progression, providing potential targets for new biomarkers and for targeted therapeutic intervention. Indeed, changes in the expression of members of the cAMP-degrading PDE4 family appear to be associated with a number of different diseases, including stroke, acrodysostosis, schizophrenia, and COPD. It has recently been shown that down-regulation of a particular isoform of PDE4 (PDE4D7) can affect prostate cancer (see, for example, Böttcher R. et al., "Human phosphodiesterase 4D7 (PDE4D7) expression is increased in TMPRSS2-ERG positive primary prostate cancer and independently adds to a reduced risk of post-surgical disease progression", British Journal of Cancer, Vol. 113, No. 10, pages 1502-1511, 2015). The PDE4D7 isoform is the so-called long isoform because it contains both the regulatory domains of UCR1 and UCR2. UCR1 is found in long, but not short, isoforms of PDE4 and is regulated by various protein kinases, including PKA and MK2, and also determines the functional outcome of catalytic unit phosphorylation by ERK. Functionally, it provides part of the cellular cAMP desensitization system and provides crosstalk between signaling pathways that lead to ERK and AMPK activation, for example.

New evidence demonstrates that human helicases such as DHX9 can act as hubs in cellular networks to coordinate cellular responses to stress to maintain cellular homeostasis for survival or initiate cell death. Interestingly, a number of human cancer cell lines have recently been shown to be susceptible to downregulation of DHX9 helicase activity in vitro and in vivo. Profiling gene expression of tumor cells after DHX9 downregulation revealed deregulation of a number of pathways involved in tumor development or progression. In addition, DHX9 has recently been described as a major player in the elimination of R-loop-associated DNA damage, suggesting the maintenance of genomic stability. Disruption of this process can lead to the development and progression of cancer due to rearrangements at the genomic level, a phenomenon that has been demonstrated in many cancers. These recent results are largely supported by our data showing a significant association of DHX9 gene expression with time to prostate cancer recurrence in a cohort of clinical prostate cancer patients with prospective follow-up (unpublished data).

The present inventors have identified and confirmed the interaction between PDE4D7 and DHX9 helicase in prostate cancer cells. In addition, the N-terminal domain of DHX9 can be shown to be phosphorylated in vitro by cAMP-dependent protein kinase A (PKA). In this regard, the question arises whether this post-translational modification under the action of PKA has any regulatory effect on the activity or cellular localization of DHX9 and whether interaction with PDE4D7 affects phosphorylation by regulating cyclic AMP in the DHX9 microdomain. Thus, PDE4D7, as a recently identified interaction partner with DHX9, may play a critical role in direct or indirect control of DHX9 function in the development of prostate cancer.

Additional molecular information elucidating the biology of the disease is obtained by determining a preoperative predictive risk score for a subject with prostate cancer based on gene expression profiles for both PDE4D7 and DHX9. The predictive power of PDE4D7 and DHX9 is used in a patient's preoperative risk assessment by determining a preoperative predictive risk score that is based on gene expression profiles for both PDE4D7 and DHX9. This may provide improved stratification of the subject in the preoperative setting, which may lead to better decisions about primary treatment. For example, preoperative predictive risk assessment may provide better guidance on the choice of active surveillance versus active intervention, such as radical prostatectomy, for certain patient subpopulations with prostate cancer.

The term "phosphodiesterase 4D7" or "PDE4D7" refers to splicing variant 7 of the human phosphodiesterase PDE4D, i.e. the human phosphodiesterase PDE4D7 gene, for example, the sequence defined in the NCBI reference sequence: NM_001165899.1, in particular the nucleotide sequence specified in SEQ ID NO:19, which corresponds to the sequence of the above NCBI reference sequence of the PDE4D7 transcript, and also refers to the corresponding amino acid sequence, for example, specified in SEQ ID NO:20, which corresponds to the protein sequence defined in the protein reference sequence under NCBI accession number NP_001159371 .1 encoding the PDE4D7 polypeptide. The term "phosphodiesterase 4D7" or "PDE4D7" also refers to an amplicon that can be generated using the primer pair PDE4D7_forward (SEQ ID NO:21) and PDE4D7_reverse (SEQ ID NO:22) and can be detected using the probe SEQ ID NO: 23.

The PDE4D7 polypeptide can also be detected using the primer pair PDE4D7-2_forward (SEQ ID NO:24) and PDE4D7_reverse (SEQ ID NO:25) and can be detected using the probe SEQ ID NO:26.

The term "PDE4D7" also includes nucleotide sequences showing a high degree of homology to PDE4D7, e.g., nucleic acid sequences that are at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94% , 95%, 96%, 97%, 98% or 99% identical to the sequence indicated in SEQ ID NO:19, or amino acid sequences that are at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the sequence specified in SEQ ID NO:20, or nucleic acid sequences encoding amino acid sequences that are at least 75% , 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the sequence specified in SEQ ID NO:20 or amino acid sequences encoded by nucleic acid sequences that are at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical the sequence shown in SEQ ID NO:19.

The term "DExH-box-helicase 9" or "DHX9" refers to the human DExH-box-helicase 9 gene, for example, the sequence defined in the NCBI reference sequence: NM_001357.4 (Ensembl: ENSG00000135829), in particular, the nucleotide sequence , specified in SEQ ID NO:74, which corresponds to the sequence of the above reference sequence of the NCBI transcript of DHX9, and also refers to the corresponding amino acid sequence, for example, specified in SEQ ID NO:75, which corresponds to the protein sequence defined in the reference sequence of the protein at number Access NCBI NP_001348.2 encoding the DHX9 polypeptide.

The term "DHX9" also includes nucleotide sequences showing a high degree of homology to DHX9, e.g., nucleic acid sequences that are at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94% , 95%, 96%, 97%, 98% or 99% identical to the sequence indicated in SEQ ID NO:74, or amino acid sequences that are at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the sequence specified in SEQ ID NO:75, or nucleic acid sequences encoding amino acid sequences that are at least 75% , 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the sequence specified in SEQ ID NO:75 or amino acid sequences encoded by nucleic acid sequences that are at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical the sequence indicated in SEQ ID NO:74.

The term "biological sample" or "subject-derived sample" refers to any biological material obtained by suitable methods known to one of skill in the art from a subject, such as a patient suffering from prostate cancer. The biological sample used can be collected in a clinically acceptable manner, for example, in such a way that nucleic acids (particularly RNA) or proteins are preserved.

The biological sample(s) may include tissue and/or body fluid such as, but not limited to, blood, sweat and urine. In addition, the biological sample may contain a cell extract derived from a population of cells, or a population of cells, including an epithelial cell such as a cancerous epithelial cell or an epithelial cell derived from a tissue suspected to be cancerous. The biological sample may contain a population of cells derived from glandular tissue, for example, the sample may be obtained from the prostate gland of a male subject. In addition, if necessary, cells can be purified from the obtained tissues and body fluids, and then used as a biological sample. In some embodiments, the sample may be a tissue sample, a urine sample, a urine sediment sample, a blood sample, a saliva sample, a semen sample, a sample including circulating tumor cells, extracellular vesicles, a sample containing exosomes secreted by the prostate, or cell lines, or a line of cancer cells.

In one particular embodiment, biopsy or resection specimens may be obtained and/or used. Such samples may include cells or cell lysates.

It is also possible that the contents of the biological sample undergo an enrichment step. For example, the sample can be brought into contact with ligands specific to the cell membrane or organelles of certain cell types, such as prostate cells, functionalized, for example, with magnetic particles. The material concentrated with magnetic particles can subsequently be used for the detection and analysis steps described above or below in this application.

In addition, cells, such as tumor cells, can be enriched by methods of filtering liquid or liquid samples, such as blood, urine, etc. Such filtration methods can also be combined with enrichment steps based on ligand-specific interactions, as described herein above.

The term "prostate cancer" refers to prostate cancer in the male reproductive system that occurs when prostate cells mutate and multiply uncontrollably. Typically, prostate cancer is associated with elevated prostate-specific antigen (PSA) levels. According to one implementation variant of the present invention, the term "prostate cancer" refers to cancer showing PSA levels above 4.0. According to another embodiment of the present invention, the term refers to cancer showing PSA levels above 2.0. The term "PSA level" refers to the concentration of PSA in the blood in ng/ml.

The term "non-progressive prostate cancer" means that the individual's sample does not show parameter values indicative of "biochemical relapse" and/or "clinical relapse".

The term "progressive prostate cancer" means that a sample of an individual shows parameter values indicative of "biochemical relapse" and/or "clinical relapse".

The term "biochemical recurrence" usually refers to recurrent biological values of elevated PSA indicating the presence of prostate cancer cells in the sample. However, other markers can also be used that can be used to detect the presence or that raise the suspicion of such presence.

The term "clinical recurrence" refers to the presence of clinical signs indicative of the presence of tumor cells when measured, for example, using in vivo imaging.

As used herein, the term "predicting prostate cancer" refers to predicting the course or outcome of a diagnosed or detected prostate cancer, for example, over a period of time, during treatment, or after treatment. The term also refers to determining the chance of surviving or recovering from a disease, as well as predicting the expected duration of a subject's survival. The prognosis may include, in particular, establishing the likelihood of the subject surviving for some period of time in the future, for example, 6 months, 1 year, 2 years, 3 years, 5 years, 10 years, or any other period of time.

Preferably, the gene expression profile for PDE4D7 and the gene expression profile for DHX9 are combined with a regression function that has been derived from a population of prostate cancer subjects.

Regression analysis helps to understand how the typical value of the dependent variable (or "criterion variable") changes when any of the independent variables varies, while the other independent variables remain fixed. This relationship between the dependent variable and the independent variables is captured in a regression function that can be used to predict the dependent variable given the values of the independent variables. The dependent variable can be, for example, a binary variable such as biochemical recurrence within 5 years of surgery. In this case, the regression is a logistic regression based on the logit function of the explanatory variables, which in this case include or consist of the gene expression profile for PDE4D7 and the gene expression profile for DHX9. Using the regression function, it is possible to improve the prognosis, for example, of the risk of biochemical recurrence within 5 years after surgery.

Preferably, the method further comprises:

suggesting a primary treatment for a subject based on a preoperative predictive risk assessment, said primary treatment being selected from the group consisting of: (i) at least partial prostatectomy; (ii) active therapy selected from radiation therapy, hormonal therapy, chemotherapy, and combinations thereof; and (iii) active surveillance.

Various national and international guidelines recommend different types of treatment for a subject suffering from prostate cancer, depending on the risk of future disease progression and life expectancy. For example, active surveillance (AS) is usually recommended for men at very low and low risk of developing prostate cancer, while radical prostatectomy may be indicated for those at high risk of developing cancer. However, known clinical risk factors do not effectively determine the intensity of the disease or its aggressiveness for all patients. For example, in the National Comprehensive Cancer Network (NCCN) very low and low risk groups, there is a significant subset of the patient population with a 10 to 25% risk of cancer recurrence after initial treatment. It is also known that in the intermediate risk group there is a subpopulation with a low risk of biochemical progression. For certain subpopulations of patients with prostate cancer, better recommendations, such as whether to choose active surveillance or active intervention, such as radical prostatectomy, can be made by basing the initial treatment offered to the subject on a preoperative predictive risk assessment.

In addition, preferably the method includes:

normalization of the gene expression profile for PDE4D7 and/or the gene expression profile for DHX9 in relation to one or more reference genes selected from the group consisting of: Homo sapiens hypoxanthine phosphoribosyltransferase 1 (HPRT1), tubulin alpha-1b (TUBA1B), a member of RNA- the Homo sapiens pumilio (PUM1) binding family; and the Homo sapiens TATA box-binding protein (TBP), wherein said preoperative predictive risk score is determined based on the normalized gene(s) expression profile(s).

The variability in determining the preoperative predictive risk score can be reduced by normalizing the expression profile(s) of the gene(s) against one or more reference genes and by determining the preoperative predictive risk score based on the normalized expression profile(s) of the gene(s) . This allows differentiation between actual variations in gene expression profiles and variations due to measurement methods. In this regard, HPRT1, TUBA1B, PUM1 and TBP have been found to be particularly well suited as reference genes for normalizing the expression profile(s) of the PDE4D7 and/or DHX9 gene(s).

The expression profile(s) of the gene(s) can be determined by detecting mRNA expression using one or more primers and/or probes and/or one or more sets thereof. Moreover, the expression profile(s) of the gene(s) can be determined using a method based on amplification and/or microarray analysis and/or RNA sequencing. Determining the expression profile(s) of the gene(s) may include performing a quantitative real-time polymerase chain reaction (RT-qPCR) on RNA isolated from a biological sample. In other embodiments, the expression profile(s) of the gene(s) are determined by RNA sequencing, standard PCR (using, for example, gel electrophoresis endpoint analysis), or multiplex PCR. In the case of RT-qPCR, determining the expression profile(s) of the gene(s) may include determining the cycle threshold (Ct) value for PDE4D7 and/or DHX9 and each of one or more reference genes. PCR can be performed using at least one primer and/or probe for measuring a reference gene selected from HPRT1, TUBA1B, PUM1 and TBP.

Preferably one or more reference genes contain at least two or at least three or all of HPRT1, TUBA1B, PUM1 and TBP.

Other reference genes that can additionally or alternatively be used to normalize the expression profile(s) of the PDE4D7 and/or DHX9 gene(s) include: actin mRNA, beta, (ACTB) Homo sapiens; mRNA of the acid ribosomal phosphoprotein P0 60S (RPLP0) Homo sapiens; polypeptide A, 220 kDa, polymerase (RNA) II (DNA directed) (POLR2A); beta-2 microglobulin (B2M); and aminolevulinate delta synthase (ALAS-1).

In some preferred embodiments, the normalized expression profile(s) of the gene(s) for PDE4D7 and/or DHX9 are converted to a predetermined range of values, e.g. 1 to 5, using the appropriate function(s) transformations. Preferably, the transformation function(s) has been(s) derived from the expression profile(s) of the gene(s) normalized(s) in an appropriate manner, e.g., the expression profile(s) of the gene(s) normalized(s) with respect to to all of HPRT1, TUBA1B, PUM1 and TBP, for PDE4D7 and/or for DHX9, for biological samples obtained from a population of prostate cancer subjects (which may preferably be the same population as the population used to obtain the function regression). In one preferred embodiment, the transform function may be a linear transform that transforms the normalized gene expression profile into a predetermined range of values. Such a transformation can be determined by looking at the frequency distribution of normalized gene expression profile values, for example, for PDE4D7 (or for DHX9) for biological samples from a population of prostate cancer subjects, and by determining a transformation that transforms the frequency distribution into the target range. With such a transform function, the expression profile of a gene can be expressed in a user-friendly way, for example, in a small range of positive values. This is similar to other categories used in everyday clinical practice, such as histopathological classification (Gleason) or MRI multiparametric radiological evaluation (PIRADS).

According to one specific implementation variant, the transformed normalized gene expression profile is determined as follows:

TNGEP = (((GENE_norm + A)*B)+1), (1)

where "TNGEP" is the transformed normalized gene expression profile, "GENE_norm" is the normalized value of the PDE4D7 or DHX9 gene expression profile, and A and B are variables and appropriately selected to display "GENE_norm" in the target value range.

Particularly preferably, the determination of the gene expression profile for PDE4D7 and/or the gene expression profile for DHX9 comprises performing RT-qPCR on RNA isolated from the biological sample(s), wherein the Cq value is determined for PDE4D7 and/or DHX9 and for each of one or more reference genes, wherein said determination of the preoperative risk score comprises normalizing the Cq value for PDE4D7 and/or DHX9 using the Cq value for each of the one or more reference genes and calculating the preoperative risk score based on the normalized value(s) Cq.

For example, a normalized Cq value for PDE4D7 can be obtained by applying the following:

N(Cq _PDE4D7 ) = Mean(Cq _{ref_genes} ) – (Cq _PDE4D7 ), (2)

where N(Cq _PDE4D7 ) is the normalized value of the gene expression profile (quantification cycle, Cq) PDE4D7, Mean(Cq _{ref_genes} ) is the arithmetic mean of the PCR Cq values of one or more reference genes, and Cq _PDE4D7 is the PCR Cq value for PDE4D7.

Similarly, the normalized Cq value for DHX9 can be obtained by applying the following:

N(Cq _DHX9 ) = Mean(Cq _{ref_genes} ) – (Cq _DHX9 ), (3)

where N(Cq _DHX9 ) is the normalized value of the gene expression profile (quantification cycle, Cq) of DHX9, Mean(Cq _{ref_genes} ) is the arithmetic mean of the PCR Cq values of one or more reference genes, and Cq _DHX9 is the PCR Cq value of DHX9.

According to a further aspect of the present invention, a diagnostic kit is provided, comprising:

at least one primer and/or probe for profiling the expression of a gene for variant 7 of phosphodiesterase 4D (PDE4D7) in a biological sample obtained from a subject suffering from prostate cancer;

at least one primer and/or probe for profiling gene expression for DExH box helicase 9 (DHX9) in the same or a different biological sample obtained from a subject; And

optionally at least one primer and/or probe for gene expression profiling for one or more reference genes selected from the group consisting of: Homo sapiens hypoxanthine phosphoribosyltransferase 1 (HPRT1), tubulin alpha-1b (TUBA1B), RNA-binding member Homo sapiens pumilio (PUM1) family and Homo sapiens TATA box-binding protein (TBP);

said diagnostic kit further comprising instructions for calculating a preoperative predictive risk score based on a gene expression profile for PDE4D7 and a gene expression profile for DHX9, said instructions being stored in a computer program product which, when executed by a computer, performs a method comprising:

determining a preoperative predictive risk score for a subject based on a gene expression profile for PDE4D7 and a gene expression profile for DHX9, optionally said method comprising:

normalization of the gene expression profile for PDE4D7 and/or the gene expression profile for DHX9 in relation to one or more reference genes,

wherein said preoperative predictive risk score is determined based on the normalized expression profile(s) of the gene(s).

Preferably, the instructions for calculating the preoperative predictive risk include instructions for combining the gene expression profile for PDE4D7 and the gene expression profile for DHX9 with a regression function that was derived from a population of prostate cancer subjects.

According to a further aspect of the present invention, the use of a diagnostic kit as defined in claim 7 or 8 is provided in a method for preoperative risk stratification in a subject suffering from prostate cancer.

According to a further aspect of the present invention, there is provided the use of a gene expression profile for phosphodiesterase 4D variant 7 (PDE4D7) and a gene expression profile for DExH-box helicase 9 (DHX9) in preoperative risk stratification in a subject suffering from prostate cancer, comprising:

determining the gene expression profile for PDE4D7 in a biological sample obtained from the subject,

determining the gene expression profile for DHX9 in the same or different biological sample obtained from the subject, and

determining a preoperative predictive risk score for the subject based on the gene expression profile for PDE4D7 and the gene expression profile for DHX9.

Preferably, the gene expression profile for PDE4D7 and the gene expression profile for DHX9 are combined with a regression function that has been derived from a population of prostate cancer subjects.

According to a further aspect of the present invention, there is provided a computer program product comprising instructions that, when executed by the computer, cause the computer to perform a method comprising:

determining a preoperative predictive risk score for a subject suffering from prostate cancer based on the gene expression profile for phosphodiesterase 4D variant 7 (PDE4D7) and the gene expression profile for DExH-box helicase 9 (DHX9),

wherein said gene expression profile for PDE4D7 was determined in a biological sample obtained from a subject, and the gene expression profile for DHX9 was determined in the same or a different biological sample obtained from a subject.

Preferably, the gene expression profile for PDE4D7 and the gene expression profile for DHX9 are combined with a regression function that has been derived from a population of prostate cancer subjects.

In further preferred embodiments, the subject's postoperative predictive risk score is determined based on the preoperative predictive risk score and the postoperative CAPRA-S clinical risk score.

Preferably, the preoperative predictive risk score and the postoperative CAPRA-S clinical risk score are combined with a regression function that has been derived from a population of subjects with prostate cancer.

Said additional preferred embodiments can be carried out using the method of claim 1, the diagnostic kit of claim 7, the use of said diagnostic kit of claim 9, the use of a gene expression profile of claim 10, and the computer program of claim 12.

It should be understood that the method of claim 1, the diagnostic kit of claim 7, the use of said diagnostic kit of claim 9, the use of the gene expression profile of claim 10, and the computer program of claim 12 have similar and/or identical preferred embodiments. , in particular as defined in the dependent claims.

It should be understood that a preferred embodiment of the present invention may also be any combination of the dependent claims or the above embodiments with the corresponding independent claim.

These and other aspects of the present invention will be understood on the basis of and explained with reference to the implementation options described in this application below.

BRIEF DESCRIPTION OF THE DRAWINGS

On the following drawings:

On FIG. 1 schematically and by way of example shows a flowchart of an embodiment of a method for preoperative risk stratification in a subject suffering from prostate cancer,

On FIG. 2 shows the results of ROC-curve analysis of biochemical recurrence (BCR) for 5 years after surgery for prostate cancer in a cohort of patients with RP.

On FIG. 3 shows the results of the ROC curve analysis of clinical recurrence (CR) over 10 years after prostate cancer surgery in the RP cohort.

On FIG. 4 shows the results of the analysis of the ROC curve of specific death from prostate cancer over 10 years after surgery for prostate cancer in a cohort of patients with RP.

DETAILED DESCRIPTION OF THE INVENTION

Overview of preoperative risk stratification

On FIG. 1 schematically and exemplarily shows a flowchart of an embodiment of a method for preoperative risk stratification in a subject suffering from prostate cancer.

The method starts at step S100.

In step S102, one or more biological samples are obtained from each of the first group of patients (subjects) diagnosed with prostate cancer. Preferably, prostate cancer monitoring for these prostate cancer patients was performed for a period of time, such as at least one year, or at least two years, or about five years, after the biological sample was obtained.

At step S104, gene expression profiles for PDE4D7 and for DHX9 are obtained for the same or different biological samples obtained from each of the first group of patients, for example, by performing RT-qPCR (quantitative real-time PCR) on RNA isolated from each biological sample. Exemplary gene expression profiles include the level of expression (eg, value) for PDE4D7 and for DHX9, which can be normalized using value(s) for each of a set of reference genes such as HPRT1, TUBA1B, PUM1 and/or TBP. In one embodiment, the PDE4D7 and DHX9 gene expression profile value is normalized to one or more reference genes selected from the group consisting of HPRT1, TUBA1B, PUM1, and TBP, e.g., at least one or at least two, or at least at least three, or preferably all of these reference genes.

In step S106, a regression function is determined to assign a preoperative predictive risk score based on the gene expression profiles for PDE4D7 and for DHX9 obtained from at least some of the biological samples obtained from the first group of patients and the corresponding monitoring results. According to one possible implementation variant, the normalized expression profile(s) of the gene(s) for PDE4D7 and/or DHX9 is first converted to a predetermined range of values, such as the range from 1 to 5 mentioned above, using the appropriate function(s) ( functions) transformations. As mentioned above, such a transformation can be determined by considering the frequency distribution of normalized values of the gene expression profile, for example, for PDE4D7 (or for DHX9) for biological samples from a population of subjects suffering from prostate cancer (in this application, the first group of patients) and by defining a transformation that transforms the frequency distribution into the target range. According to one specific implementation variant, the normalized (s) transformed (s) value (s) of the gene expression profile is determined as indicated in equation (1) above.

In step S108, one or more biological samples are obtained from a patient (subject or individual). The patient may be a new patient or one from the first group.

In step S110, gene expression profiles for PDE4D7 and DHX9 are obtained, for example by performing PCR on a biological sample. In one embodiment, the values of the PDE4D7 and DHX9 gene expression profiles are normalized to one or more reference genes selected from the group consisting of HPRT1, TUBA1B, PUM1, and TBP, e.g., at least one or at least two, or at least at least three, or preferably all of these reference genes. This step is essentially the same as step S104. Moreover, the normalized expression profile(s) of the gene(s) for PDE4D7 and/or DHX9 can be pre-transformed as described in relation to step S106.

Other reference genes that may additionally or alternatively be used in steps S104 and S110 include: actin mRNA, beta, (ACTB) Homo sapiens; mRNA of the acid ribosomal phosphoprotein P0 60S (RPLP0) Homo sapiens; polypeptide A, 220 kDa, polymerase (RNA) II (DNA directed) (POLR2A); beta-2 microglobulin (B2M); and aminolevulinate delta synthase (ALAS-1).

In step S112, a preoperative predictive risk score for the patient is determined using a regression function based on the gene expression profiles for PDE4D7 and DHX9. This will be described in more detail below in the description. To make the preoperative predictive score intuitive to the user, it can also be defined so that its values fall within a predetermined range of values, for example, the range from 1 to 5. This can be achieved either with the regression function itself or with using an appropriate post-transformation that converts the output values of the regression function to the target range of values. As already mentioned, this is similar to other categories used in everyday clinical practice, such as histopathological classification (Gleason) or MRI multiparametric radiological evaluation (PIRADS).

At S114, a recommendation for therapy may be provided to, for example, a patient or his or her caregiver, physician, or other healthcare professional based on a preoperative predictive risk assessment. To this end, the preoperative predictive risk score may be assigned to one group from a predetermined set of risk groups based on the value of the preoperative predictive risk score. The provision of a therapy recommendation may include one or more of the following: a) suggesting therapy for a patient based on an assigned risk group, with at least two of the risk groups associated with different therapies, b) calculating a patient's risk of disease progression to or after prostate surgery; and c) calculating a predictive response to therapy for a patient before or after prostate surgery. Exemplary therapies include at least partial prostatectomy, active therapy selected from radiation therapy, chemotherapy, and combinations thereof, and observation only, i.e., without performing prostatectomy or active therapy (i.e., active observation).

The method ends at S116.

Each of the risk groups can be associated with the corresponding proposed therapy, which differs in its aggressiveness. Each proposed therapy can be based on the results of patients in the first group who were assigned to this risk group, and is a therapy that is predicted to provide the least aggressive therapy that does not exceed the threshold clinical risk for developing prostate cancer. In some cases, this allows the new patient to be placed at risk associated with a less aggressive proposed therapy than may be the case with other risk profiling methods such as using the Gleason score, NCCN risk categories, or preoperative CAPRA assessment.

In one embodiment, gene expression profiles in steps S104 and S110 are determined by detecting mRNA expression using one or more primers and/or probes and/or one or more sets thereof.

Detailed descriptions of PDE4D7, DHX9, and one or more reference genes, including their transcript identifier (NCBI RefSeq) and corresponding amino acid sequences for the primer and probe pair, are shown in TABLE 1. This table also shows the sense primer and antisense primer for each gene, and also the probe sequence that specifically binds to the amplicon.

TABLE 1: Exemplary primer and probe nucleic acid sequences

Gene name Reference. afterbirth. NCBI of exemplary protein Exemplary protein access number Sense primer Antisense primer Probe sequence PDE4D7 NM_001165899.1 (SEQ ID NO:19) NP_001159371.1 (SEQ ID NO:20) GAACATTCAACGACCAACCA (SEQ ID NO:21) TGCCATTGTCCACATCAAAAA (SEQ ID NO:22) CTGCCGCTGATTGCTATCACTTCTGCA (SEQ ID NO:23) CGCTGATTGCTATCACTTCTGC (SEQ ID NO:24) GTCGTTGACTGTGGACAAAATTTG (SEQ ID NO:25) TTCCCTTGGATCCCATGACCAGCCCATAAGGGAA (SEQ ID NO:26) DHX9 NM_00135
7.4 (SEQ ID NO:74) NP_00134 8.2 (SEQ ID NO:75) Must be custom designed Must be custom designed Must be custom designed

HPRT1 NM_00019
4.2 (SEQ ID NO:34) NP_000185.1 (SEQ ID NO:35) GAGGATTTGGAAAGGGTGTTTATT
(SEQ ID NO:36) ACAGAGGGCTACAATGTGATG
(SEQ ID NO:37) ACGTCTTGCTCGAGATGTGATGAAGG
(SEQ ID NO:38) TUBA1B NM_00608
2.2 (SEQ ID NO:39) NP_006073.2 (SEQ ID NO:40) TGACTCCTTCAACACCTTCTC (SEQ ID NO:41) TGCCAGTGCGAACTTCAT (SEQ ID NO:42) CCGGGCTGTGTTTGTAGACTTGGA (SEQ ID NO:43) PUM1 NM_001020658.1 (SEQ ID NO:44); NM_01467
6.2 (SEQ ID NO:45) NP_001018494.1 (SEQ ID NO:46); NP_05549
1.1 (SEQ ID NO:47) GCCAGCTTGTCTTCAATGAAAT (SEQ ID NO:48) CAAAGCCAGCTTCTGTTCAAG (SEQ ID NO:49) ATCCACCATGAGTTGGTAGGCAGC (SEQ ID NO:50) TBP NM_00319
4.4 (SEQ ID NO:51) NP_00318
5.1 (SEQ ID NO:52) GCCAAGAAGAAAGTGAACATCAT
(SEQ ID NO:53) ATAGGGATTCCGGGAGTCAT (SEQ ID NO:54) TCAGAACAACAGCCTGCCACCTTA (SEQ ID NO:55) ACTB NM_00110
1.3 SEQ ID NO:56) NP_00109
2.1 (SEQ ID NO:57) CCAACCGCGAGAAGATGA (SEQ ID NO:58) CCAGAGGCGTACAGGGATAG (SEQ ID NO:59) CCATGTACGTTGCTATCCAGGCT (SEQ ID NO:60) RPLP0 NM_00100
2.3 (SEQ ID NO:61) NP_44450
5.1/NP_000993.1 (SEQ ID NO:62/63) TAAACCCTGCGTGGCAAT (SEQ ID NO:64) ACATTTCGGATAATCATCCAATAGTTG (SEQ ID NO:65) AAGTAGTTGGACTTCCAGGTCGCC (SEQ ID NO:66) ALAS-1 NM_00068
8.5/NM_199166.2 (SEQ ID NO:67/68) NP_00067
9.1/NP_954635.1 (SEQ ID NO:69/70) AGCCACATCATCCCTGT SEQ ID NO:71) CGTAGATGTTATGTCTGCTCAT (SEQ ID NO:72) TTTAGCAGCATCTGCAACCCGC (SEQ ID NO:73)

Instead of using RT-qPCR, gene expression profiles for PDE4D7 and/or DHX9 in steps S104 and S110 can be determined in other embodiments using other methods, for example, by performing next generation RNA sequencing (NGS RNAseq) according to standard methods (Illumina , Inc.). In this case, the described transformations to a predetermined range of values can also be performed on the expression profile(s) of the gene(s) provided(s) by NGS RNAseq.

To investigate the predictive power of PDE4D7 and DHX9 in preoperative patient risk assessment, the correlation with disease recurrence was studied.

A combined model based on PDE4D7 and DHX9 gene expression profiles was developed in the surgical cohort and this model was tested for various prospective clinical outcomes. The results indicate that the combination of gene expression profiles for both PDE4D7 and DHX9 in improved preoperative predictive risk assessment may provide improved patient stratification to optimize primary treatment decisions.

EXAMPLES

Patient cohort and samples

A cohort of radical prostatectomy (RP) patients whose demographic characteristics are shown in TABLE 2 was used. who were operated on consecutively between 2000 and 2004 in one large clinical center in Germany.

TABLE 2: Demographic Characteristics of the Radical Prostatectomy (RP) Patient Cohort

Operation: 2000-2004 Parameter RP cohort (#575) Demographic and clinical characteristics
Range (median) Age (at RP) 41.3-79.2 (62.7) Preoperative PSA 0.18-120.0 (7.1) Percentage of tumor on biopsy 0.2-80.0 (10.3) Prostate volume 9-244 (42) PSA density 0.01-24.0 (0.18) CAPRA-S risk category
Number of patients
(percentage) Low risk (CAPRA-S 0-2) 275 (47.8%) Intermediate risk (CAPRA-S 3-5) 220 (38.3%) High risk (CAPRA-S>5) 80 (13.9%) Postoperative pathology
Number of patients
(percentage) Gleason Pathology 3+3 (GG1) 190 (33%) Gleason Pathology 3+4 (GG2) 288 (50.1%) Gleason Pathology 4+3 (GG3) 73 (12.7%) Gleason pathology >= 4 + 4 (≥GG4) 24 (4.2%) pT2 331 (57.6%) pT3 244 (42.4%)

pT4 0 (0%) Positive Surgical Margins 211 (36.7%) Penetration beyond the capsule (= T3a) 151 (26.3%) Positive invasion of the seminal vesicles 95 (16.5%) Positive lymph node invasion 20 (3.5%) Observation
Months Average 104.3 Median IQR 120 Exodus
Number of events/
total number of patients
(percentage) <5 years BCR 184/512 (35.9%) <10 years BCR 228/428 (53.3%) <5 years CR 49/503 (9.7%) <10 years CR 64/356 (18.0%) Salvation Therapy
Number of events/
total number of patients
(percentage) <5 years SRT 141/506 (27.9%) <10 years SRT 178/405 (44.0%) <5 years SADT 79/498 (15.9%) <10 years SADT 118/370 (31.9%) Mortality
Number of events/
total number of patients
(percentage) <5 years PCSS 14/483 (2.9%) <10 years PCSS 26/321 (8.1%) <5 years OS 27/496 (5.4%) <10 years OS 54/349 (15.5%)

For patient age, preoperative PSA, percentage of tumor at biopsy, prostate volume, and PSA density, the minimum and maximum values in the cohort are shown, while the median values are shown in brackets. For CAPRA-S risk categories, the number of patients and the percentage in the risk group are shown. Postoperative pathology is represented by Gleason pathology scores and groups classified by Gleason scale, stages of pathology, status of surgical margin after prostatectomy, status of tumor invasion of seminal vesicles and pelvic lymph nodes (by number and percentage of patients). In this regard, it should be noted that penetration beyond the capsule was not presented as a main parameter, but was obtained based on the pathology stage of pT3a. Follow-up shows mean and median follow-up in months after surgery for all patients. The outcome category illustrates pooled data on biochemical recurrence (BCR) at 5 and 10 years and clinical recurrence before metastasis (CR) after primary surgical treatment. The treatment category lists cumulative data for initiation of salvage radiotherapy (SRT) or salvage therapy by androgen depletion (SADT) during 5 and 10 years after surgery. Mortality is shown as prostate cancer specific survival (PCSS) as well as overall survival (OS). For all outcomes, the number of men who survived the outcome is shown per total number of men with corresponding 5- or 10-year follow-up periods, with the percentage of events shown in parentheses.

Laboratory methods

All laboratory methods used, including oligonucleotide primers and probes for RT-qPCR (quantitative real-time PCR), RNA extraction and quality control, as well as inclusion / exclusion of samples from statistical analysis, were similar to those described previously in Böttcher R. et al. or, in the case of DHX9, were based on next generation RNA sequencing (NGS RNAseq) according to standard methods (Illumina, Inc.). The primers and probes used for RT-qPCR to measure genes of interest as well as reference genes are also shown in TABLE 1. In the case of NGS RNAseq, the resulting FastQ files were matched to the human genome and processed according to standard methods. For each gene, the expression value of the gene was obtained as TPM (transcript per million kb; see http://www.rna-seqblog.com/rpkm-fpkm-and-tpm-clearly-explained (accessed 7 October 2018).

RESULTS

Modeling PDE4D7 and DHX9 Expression Using Logistic Regression

Expression values for PDE4D7 and DHX9 were used in logistic regression modeling to create a combined model. As shown in TABLE 1 below, biochemical recurrence (BCR) within 5 years of prostate cancer surgery was taken as the dependent variable in a subcohort of 481 patients (169 events; 35.14%) of the RP patient cohort with complete 5- summer outcome stories. The regression function logit(p) was converted to p=1/(1+e^(-logit(p)) to calculate the probability p that an individual patient will experience a biochemical relapse within 5 years of surgery. TABLES 2-6 show results of modeling using logistic regression to combine PDE4D7 and DHX9 expression values TABLE 1 describes the input data for modeling using logistic regression in terms of cohort size (#481) and the number of positive cases with biochemical relapse (BCR) over 5 years and negative cases without BCR over 5 years.TABLE 2 provides information on the model fit and TABLE 3 presents the coefficients (or weights) of the regression model with associated statistics.TABLE 4 provides an overview of the odds ratios for PDE4D7 and DHX9, and TABLE 5 and 6 shows the classification table and ROC curve analysis data for the "PDE4D7 and DHX9" regression model.

TABLE 1: Simulation input data using logistic regression.

Dependent variable Y Biochemical relapse (BCR) within 5 years Method Input Sample size 481 positive cases ^a 169 (35.14%) Negative cases ^b 312 (64.86%)

^a BCR within 5 years = 1

^b BCR within 5 years = 0

TABLE 2: General approximation of the model.

Zero Model - 2 Log Plausibility 623.645 Full Model - 2 Log Plausibility 557.901 Chi-square 65.744 D.F. 2 Significance level P<0.0001 R ² according to Cox and Snell 0.1278 R²By Nagelkerke 0.1758

TABLE 3: Coefficients and standard errors.

Variable Coefficient standard error Wald (Wald) P PDE4D7 -0.98799 0.17158 33.1574 < 0.0001 DHX9 -0.75891 0.17095 19.7082 < 0.0001 Constant 7.96032 1.38277 33.1409 < 0.0001

TABLE 4: Odds ratios and 95% confidence intervals.

Variable odds ratio 95% CI PDE4D7 0.3723 0.2660-0.5212 DHX9 0.4682 0.3349-0.6545

TABLE 5: Classification table (cut-off p = 0.5).

Actual group Predicted group Correct percentage 0 1 Y=0 277 35 88.78% Y=1 116 53 31.36% Percentage of correctly classified cases 68.61%

TABLE 6: ROC curve analysis.

Area under the ROC curve (AUC) 0.713 standard error 0.0241 95% confidence interval 0.670-0.753

Validation of the Combined Model for Different Prospective Clinical Outcomes

The combined model was tested to predict various clinically relevant endpoints after surgery such as biochemical recurrence (BCR), clinical recurrence (progression to local and/or distant metastases) (CR) and specific death from prostate cancer. The predictive power of the combined model was compared with the predictive power of PDE4D7 or DHX9 alone for the same different clinically relevant endpoints.

On FIG. 2 shows the results of ROC-curve analysis of biochemical recurrence (BCR) for 5 years after prostate cancer surgery for a subcohort (#481) of patients from the RP cohort with a full 5-year follow-up (see also TABLE 7). As detailed in TABLE 8, the calculated 5-year AUC (area under the curve) was 0.659 for PDE4D7 alone, 0.624 for DHX9 alone, and 0.713 for the combined model (labeled "PDE4D7 and DHX9" in Figure 2 and TABLE 8 ). The standard error was 0.0254 (PDE4D7), 0.0280 (DHX9), and 0.0242 (PDE4D7 and DHX9), respectively, the 95% confidence interval was 0.615 to 0.702 (PDE4D7), 0.579 to 0.667 (DHX9), and 0.670 to 0.753 (PDE4D7 and DHX9), respectively. TABLE 9 shows a pairwise comparison of the ROC curves. As can be seen, PDE4D7 alone and combined model AUC tests of PDE4D7 and DHX9 are significantly different (p = 0.0022). The same is true even to a slightly greater extent for the AUC of DHX9 alone and the combined model of PDE4D7 and DHX9 (p = 0.0008).

TABLE 7: Sub-cohort.

Variable 1 PDE4D7 Variable 2 DHX9 Variable 3 PDE4D7 and DHX9 classification variable BCR for 5 years Sample size 481 positive cases ^a 169 (35.14%) Negative cases ^b 312 (64.86%)

^a BCR within 5 years = 1

^b BCR within 5 years = 0

TABLE 8: ROC curve analysis.

Variable AUC standard error ^with 95% ^CI PDE4D7 0.659 0.0254 0.615-0.702 DHX9 0.624 0.0280 0.579-0.667 PDE4D7 and DHX9 0.713 0.0242 0.670-0.753

^c DeLong ER et al., "Comparing the areas under two or more correlated receiver operating characteristic curves: a nonparametric approach", Biometrics, Vol. 44, no. 3, pages 837-845, 1988.

^b Binom

ial accurate DI

TABLE 9: Pairwise comparison of ROC curves.

PDE4D7~DHX9 The difference between areas 0.0352 standard error ^with 0.0392 95% confidence interval -0.0416-0.112 Z-score 0.899 Significance level p=0.3687 PDE4D7 ~ PDE4D7 and DHX9 The difference between areas 0.0538 standard error ^with 0.0176 95% confidence interval 0.0193-0.0883 Z-score 3.058 Significance level p = 0.0022 DHX9 ~ PDE4D7 and DHX9 The difference between areas 0.0890 standard error ^with 0.0265 95% confidence interval 0.0372-0.141 Z-score 3.365 Significance level p = 0.0008

^c DeLong ER et al.

On FIG. 3 shows the results of the analysis of the ROC curve of clinical recurrence (CR) for 10 years after surgery for prostate cancer for a subcohort (#335) of patients from a cohort of patients with RP with a full 10-year follow-up (see also TABLE 10). As detailed in TABLE 11, the calculated 10-year AUC (area under the curve) was 0.666 for PDE4D7 alone, 0.691 for DHX9 alone, and 0.748 for the combined model (labeled "PDE4D7 and DHX9" in Figure 3 and TABLE 11 ). The standard error was 0.0364 (PDE4D7), 0.0384 (DHX9), and 0.0314 (PDE4D7 and DHX9), respectively, the 95% confidence interval was 0.613 to 0.717 (PDE4D7), 0.639 to 0.740 (DHX9), 0.698 to 0.794 (PDE4D7 and DHX9), respectively. TABLE 12 shows a pairwise comparison of the ROC curves. As can be seen, PDE4D7 alone and combined model AUC tests of PDE4D7 and DHX9 are significantly different (p = 0.0039).

TABLE 10: Sub-cohort.

Variable 1 PDE4D7 Variable 2 DHX9 Variable 3 PDE4D7 and DHX9 classification variable Clinical relapse (CR) within 10 years Sample size 335 positive cases ^a 61 (18.21%) Negative cases ^b 274 (81.79%)

^a CR over 10 years = 1

^b CR over 10 years = 0

TABLE 11: ROC curve analysis.

Variable AUC standard error ^with 95% ^CI PDE4D7 0.666 0.0364 0.613-0.717 DHX9 0.691 0.0384 0.639-0.740 PDE4D7 and DHX9 0.748 0.0314 0.698-0.794

^c DeLong ER et al.

^d Binomial exact CI

TABLE 12: Pairwise comparison of ROC curves.

PDE4D7~DHX9 The difference between areas 0.0249 standard error ^with 0.0573 95% confidence interval -0.0873-0.137 Z-score 0.435 Significance level p=0.6635 PDE4D7 ~ PDE4D7 and DHX9 The difference between areas 0.0816 standard error ^with 0.0283 95% confidence interval 0.0262-0.137 Z-score 2.888 Significance level p = 0.0039 DHX9 ~ PDE4D7 and DHX9 The difference between areas 0.0567 standard error ^with 0.0361 95% confidence interval -0.0141-0.128 Z-score 1.569 Significance level p = 0.1166

^c DeLong ER et al.

On FIG. 4 shows the results of the analysis of the ROC curve of specific death from prostate cancer within 10 years after surgery for prostate cancer for a subcohort (#302) of patients from a cohort of patients with RP with a full 10-year follow-up (see also TABLE 13). As detailed in TABLE 14, the calculated 10-year AUC (area under the curve) was 0.729 for PDE4D7 alone, 0.622 for DHX9 alone, and 0.754 for the combined model (labeled "PDE4D7 and DHX9" in Figure 4 and TABLE 14 ). The standard error was 0.0588 (PDE4D7), 0.0593 (DHX9) and 0.0481 (PDE4D7 and DHX9), respectively, the 95% confidence interval was 0.676 to 0.779 (PDE4D7), 0.564 to 0.676 (DHX9) 0.702 to 0.802 (PDE4D7 and DHX9), respectively. TABLE 15 shows a pairwise comparison of the ROC curves. As can be seen, PDE4D7 and DHX9 AUCs of DHX9 alone and combined model are significantly different (p = 0.0403).

TABLE 13: Sub-cohort.

Variable 1 PDE4D7 Variable 2 DHX9 Variable 3 PDE4D7 and DHX9 classification variable Specific death from prostate cancer within 10 years Sample size 302 positive cases ^a 25 (8.28%) Negative cases ^b 277 (91.72%)

^a PCSS over 10 years = 1

^b PCSS over 10 years = 0

TABLE 14: ROC curve analysis.

Variable AUC standard error ^with 95% ^CI PDE4D7 0.729 0.0588 0.676-0.779 DHX9 0.622 0.0593 0.564-0.676 PDE4D7 and DHX9 0.754 0.0481 0.702-0.802

^c DeLong ER et al.

^d Binomial exact CI

TABLE 15: Pairwise comparison of ROC curves.

PDE4D7~DHX9 The difference between areas 0.108 standard error ^with 0.0909 95% confidence interval -0.0702-0.286 Z-score 1.188 Significance level p = 0.2349 PDE4D7 ~ PDE4D7 and DHX9 The difference between areas 0.0249 standard error ^with 0.0356 95% confidence interval -0.0448-0.0946 Z-score 0.700 Significance level p = 0.4836 DHX9 ~ PDE4D7 and DHX9 The difference between areas 0.133 standard error ^with 0.0648 95% confidence interval 0.00585-0.260 Z-score 2,050 Significance level p = 0.0403

^c DeLong ER et al.

The presented results demonstrate that the use of a combined PDE4D7 logistic regression model with DHX9 gene expression values to predict biochemical recurrence within 5 years after surgery improves the area under the curve (AUC) in the ROC analysis by 2.5% (for specific cancer death prostate as endpoint) up to 8% (for clinical recurrence as endpoint) compared with PDE4D7 alone as a prognostic marker.

Independent testing of a combined model for predicting metastases after prostate cancer surgery on an independent dataset

On FIG. 5 shows independent testing of a combined model (denoted as "PDE4D7 and DHX9" in TABLE 16) for predicting metastases after prostate cancer surgery in an independent data set (data from Taylor B.S. et al., "Integrative genomic profiling of human prostate cancer" , Cancer Cell, Vol 18, No 1, pages 11-22, 2010). As detailed in TABLE 16, the calculated AUC (area under the curve) of the metastatic class was 0.736 for PDE4D7 and DHX9. The standard error was 0.0777, the 95% confidence interval was from 0.651 to 0.809.

TABLE 16: Independent data set.

Variable PDE4D7 and DHX9 classification variable Metastasis class Sample size 129 positive cases ^a 8 (6.20%) Negative cases ^b 121 (93.8%) Disease prevalence (%) unknown

^a Metastasis class = 1

^b Metastasis class = 0

TABLE 17: ROC curve analysis.

AUC 0.736 standard error ^with 0.0777 95% confidence interval ^d 0.651-0.809 Z-score 3.029 Significance level p = 0.024

^c DeLong ER et al.

^d Binomial exact CI

TABLE 18: Youden index.

Youden index J 0.4194 Associated criterion > -1.16 Sensitivity 75.00 Specificity 66.94

Logistic regression modeling of the combined PDE4D7 and DHX9 model with postoperative clinical risk score CAPRA-S to predict postoperative metastases

The PDE4D7 and DHX9 combined model and CAPRA-S postoperative clinical risk assessment were used in additional modeling using logistic regression to generate an additional combined model (labeled "PDE4D7 and DHX9 and CAPRA-S" below). As shown in TABLE 19 below, the class of metastases after prostate cancer surgery was taken as the dependent variable in the independent data from Tayler B.S. et al. (see above). The regression function logit(p) was converted to p=1/(1+e^(-logit(p)) to calculate the probability p that an individual patient will develop metastases after surgery. TABLES 20-26 show the simulation results using logistic regression for the combination of PDE4D7 and DHX9 and the CAPRA-S score.TABLE 19 describes the input data for modeling using logistic regression in terms of sample size (#129) and the number of positive cases with metastases and negative cases without metastases.TABLE 20 provides information about the model fit, and TABLE 21 describes the coefficients (or weights) of the regression model with associated statistics.TABLE 22 provides an overview of the odds ratios for PDE4D7 and DHX9 and the CAPRA-S score, while TABLEs 23 and regression model testing (according to Hosmer and Lemeshow's test) Finally, TABLES 25 and 26 present classification table and ROC curve analysis data for the regression model "PDE4D7 and DHX9 and CAPRA-S".

TABLE 19: Simulation input data using logistic regression.

Dependent variable Y Class of metastases Method Input Sample size 129 positive cases ^a 8 (6.20%) Negative cases ^b 121 (93.8%)

^a Metastasis class = 1

^b Metastasis class = 0

TABLE 20: General approximation of the model.

Zero Model - 2 Log Plausibility 59.979 Full Model - 2 Log Plausibility 49.989 Chi-square 9.990 D.F. 2 Significance level p = 0.0068 R ² according to Cox and Snell 0.0745 R²By Nagelkerke 0.2004

TABLE 21: Coefficients and standard errors.

Variable Coefficient standard error Wald P PDE4D7 and DHX9 0.77252 0.42392 3.3208 0.0684 CAPRA-S score 0.25375 0.12488 4.1290 0.0422 Constant -2.80880 0.87165 10.3837 0.0013

TABLE 22: Odds ratios and 95% confidence intervals.

Variable odds ratio 95% CI PDE4D7 and DHX9 2.1652 0.9433-4.9699 CAPRA-S score 1.2888 1.0090-1.6463

TABLE 23: Hosmer and Lemeshow test.

Chi-square 2.9654 D.F. 8 Significance level p=0.9365

TABLE 24: Contingency table for the Hosmer and Lemeshow test.

Group Y=0 Y=1 Total Observable expected Observable expected 1 13 12.928 0 0.072 13 2 13 12.863 0 0.137 13 3 13 12.776 0 0.224 13 4 13 12.696 0 0.304 13 5 12 12.615 1 0.385 13 6 13 12.499 0 0.501 13 7 12 12.292 1 0.708 13 8 12 12.040 1 0.960 13 9 12 11.433 1 1.567 13 10 8 8.857 4 3.143 12

TABLE 25: Classification table (cut-off p = 0.5).

Actual group Predicted group Correct percentage 0 1 Y=0 120 1 99.17% Y=1 8 0 0.00% Percentage of correctly classified cases 93.02%

TABLE 26: ROC curve analysis.

AUC 0.713 standard error ^with 0.0241 95% confidence interval ^d 0.670-0.753

On FIG. Figure 6 shows ROC analysis of combined models of PDE4D7 and DHX9, as well as PDE4D7 and DHX9 and CAPRA-S for predicting metastases after prostate cancer surgery (data from Taylor B.S. et al., “Integrative genomic profiling of human prostate cancer”, Cancer Cell, Vol. 18, No. 1, pages 11-22, 2010). As detailed in TABLE 28, the calculated AUC (area under the curve) of the metastatic class was 0.736 for PDE4D7 and DHX9 alone and 0.840 for PDE4D7 and DHX9 and CAPRA-S. The standard error was 0.0777 (PDE4D7 and DHX9) and 0.0636 (PDE4D7 and DHX9 and CAPRA-S), respectively; and CAPRA-S), respectively. TABLE 29 shows a pairwise comparison of the ROC curves. As can be seen, the AUCs of PDE4D7 and DHX9 alone and in the combined model of PDE4D7 and DHX9 and CAPRA-S were significantly different (p = 0.0323), with a difference in AUC of 0.104 (or 10.4%).

TABLE 27: Independent data set.

Variable 1 PDE4D7 and DHX9 Variable 2 PDE4D7 and DHX9 and CAPRA-S classification variable Class of metastases Sample size 129 positive cases ^a 8 (6.20%) Negative cases ^b 121 (93.8%) Disease prevalence (%) unknown

^a Metastasis class = 1

^b Metastasis class = 0

TABLE 28: ROC curve analysis.

Variable AUC standard error ^with 95% CI ^d PDE4D7 and DHX9 0.736 0.0777 0.651-0.809 PDE4D7 and DHX9 and CAPRA-S 0.840 0.0636 0.765-0.899

^c DeLong ER et al.

^d Binomial exact CI

TABLE 29: Pairwise comparison of ROC curves.

PDE4D7 & DHX9 ~ PDE4D7 & DHX9 & CAPRA-S The difference between areas 0.104 standard error ^with 0.0487 95% confidence interval 0.00881-0.200 Z-score 2.141 Significance level p = 0.0323

^c DeLong ER et al.

Discussion

Treatment decisions for primary localized prostate cancer are highly dependent on a combination of risk of future disease progression and life expectancy. These data illustrate that the use of a combination of PDE4D7 and DHX9 in preoperative risk assessment improves performance compared to using PDE4D7 alone as a prognostic marker. Thus, DHX9 may enhance the predictive value of PDE4D7 in clinical predictive models for disease-specific outcomes such as postoperative progression to biochemical recurrence or clinical recurrence to metastasis, as well as to predict specific cancer survival to support treatment decisions.

Other variations of the disclosed embodiments may be understood and practiced by those skilled in the art in the practice of the claimed invention based on examination of the drawings, the disclosure, and the appended claims.

In the claims, the word "comprising" does not exclude other elements or steps, and the indefinite article ("a" or "an" in the original English text) does not exclude plurality.

One or more steps of the method illustrated in FIG. 1 may be implemented in a computer program product that may be executed on a computer. The computer program product may include a non-volatile computer-readable recording medium on which a control program is recorded (stored), such as a disk drive, a hard disk drive, or the like. Common forms of non-volatile computer-readable media include, for example, floppy disks, floppy disks, hard disks, magnetic tape or any other magnetic data storage medium, CD-ROM, DVD or any other optical media, RAM, PROM, EPROM, FLASH-EPROM or other a memory chip or cartridge or any other non-volatile medium that a computer can read and use.

Alternatively, one or more method steps may be implemented in volatile media such as a transmitted carrier wave in which the control program is implemented as a data signal using a transmission medium such as acoustic or light waves such as those generated in time of radio wave and infrared data transmissions, etc.

An exemplary method may be implemented on one or more general purpose computers, special purpose computer(s), a programmed microprocessor or microcontroller, and peripheral elements of an integrated circuit, ASIC or other integrated circuit, digital signal processor, wired electronics or logic circuit, such as a circuit with discrete elements, a programmable logic device such as a PLD, PLA, FPGA, graphics card CPU (GPU) or PAL, or the like. In general, any device capable of implementing a state machine, which in turn is capable of implementing the block diagram shown in FIG. 1 may be used to implement one or more steps of a risk stratification method for selecting therapy in a patient suffering from prostate cancer, as illustrated. It will be appreciated that while all steps of the method may be performed on a computer, in some embodiments, one or more of the steps may be performed, at least in part, manually.

Computer program instructions may also be loaded into a computer, other programmable processing device, or other devices to cause a series of operational steps to be performed on the computer, other programmable device, or other devices to provide a computer-implemented method such that the instructions that are executed on the computer or other programmable device, provide methods for implementing the functions/actions specified in this application.

Although the present invention has been described above based on the gene expression profile for PDE4D7, which may include the expression level (e.g., value) for PDE4D7, which can be normalized using the value(s) for each of the set of reference genes, the expression profile the gene may additionally include information about the expression of other PDE4D variants. For example, other PDE4D variant(s) may include one or more of PDE4D1, PDE4D2, PDE4D3, PDE4D4, PDE4D5, PDE4D6, PDE4D8, and PDE4D9. Thus, the diagnostic kit may further comprise at least one primer and/or probe for profiling gene expression for each of the other PDE4D variants in a biological sample obtained from a subject with prostate cancer. Preferably, however, only the gene expression profile for PDE4D7 is used, in particular the expression level (eg value) for PDE4D7, which can be normalized using the value(s) for each of the set of reference genes.

The term "phosphodiesterase 4D1" or "PDE4D1" refers to splicing variant 1 of the human phosphodiesterase PDE4D, i.e. the human phosphodiesterase PDE4D1 gene, for example, the sequence defined in the NCBI reference sequence: NM_001197222.1, in particular the nucleotide sequence specified in SEQ ID NO:1, which corresponds to the sequence of the above NCBI reference sequence of the PDE4D1 transcript, and also refers to the corresponding amino acid sequence, for example, specified in SEQ ID NO:2, which corresponds to the protein sequence defined in the protein reference sequence under NCBI accession number NP_001184151 .1 encoding the PDE4D1 polypeptide. The term "phosphodiesterase 4D1" or "PDE4D1" also refers to an amplicon that can be generated using the primer pair PDE1D1D2_forward (SEQ ID NO:3) and PDE1D1D2_reverse (SEQ ID NO:4) and can be detected using the probe SEQ ID NO: 5.

The term “phosphodiesterase 4D2” or “PDE4D2” refers to splicing variant 2 of the human phosphodiesterase PDE4D, i.e. the human phosphodiesterase PDE4D2 gene, for example, the sequence defined in the NCBI reference sequence: NM_001197221.1, in particular the nucleotide sequence specified in SEQ ID NO:6, which corresponds to the sequence of the above NCBI reference sequence of the PDE4D2 transcript, and also refers to the corresponding amino acid sequence, for example, specified in SEQ ID NO:7, which corresponds to the protein sequence defined in the protein reference sequence under NCBI accession number NP_001184150 .1 encoding the PDE4D2 polypeptide.

The term "phosphodiesterase 4D3" or "PDE4D3" refers to splicing variant 3 of the human phosphodiesterase PDE4D, i.e. the human phosphodiesterase PDE4D3 gene, e.g. the sequence defined in the NCBI reference sequence: NM_006203.4, in particular the nucleotide sequence specified in SEQ ID NO:8, which corresponds to the sequence of the above NCBI reference sequence of the PDE4D3 transcript, and also refers to the corresponding amino acid sequence, for example, specified in SEQ ID NO:9, which corresponds to the protein sequence defined in the protein reference sequence under NCBI accession number NP_006194 .2 encoding the PDE4D3 polypeptide.

The term "phosphodiesterase 4D4" or "PDE4D4" refers to splicing variant 4 of the human phosphodiesterase PDE4D, i.e. the human phosphodiesterase PDE4D4 gene, for example, the sequence defined in the NCBI reference sequence: NM_001104631.1, in particular the nucleotide sequence specified in SEQ ID NO:10, which corresponds to the sequence of the above NCBI reference sequence of the PDE4D4 transcript, and also refers to the corresponding amino acid sequence, for example, specified in SEQ ID NO:11, which corresponds to the protein sequence defined in the reference protein sequence under NCBI accession number NP_001098101 .1 encoding the PDE4D4 polypeptide.

The term "phosphodiesterase 4D5" or "PDE4D5" refers to splicing variant 5 of the human phosphodiesterase PDE4D, i.e. the human phosphodiesterase PDE4D5 gene, for example, the sequence defined in the NCBI reference sequence: NM_001197218.1, in particular the nucleotide sequence specified in SEQ ID NO:12, which corresponds to the sequence of the above NCBI reference sequence of the PDE4D5 transcript, and also refers to the corresponding amino acid sequence, for example, specified in SEQ ID NO:13, which corresponds to the protein sequence defined in the reference protein sequence under NCBI accession number NP_001184147 .1 encoding the PDE4D5 polypeptide. The term "phosphodiesterase 4D5" or "PDE4D5" also refers to an amplicon that can be generated using the primer pair PDE4D5_forward (SEQ ID NO:14) and PDE4D5_reverse (SEQ ID NO:15) and can be detected using the probe SEQ ID NO: 16.

The term "phosphodiesterase 4D6" or "PDE4D6" refers to splicing variant 6 of the human phosphodiesterase PDE4D, i.e. the human phosphodiesterase PDE4D6 gene, for example, the sequence defined in the NCBI reference sequence: NM_001197223.1, in particular the nucleotide sequence specified in SEQ ID NO:17, which corresponds to the sequence of the above NCBI reference sequence of the PDE4D6 transcript, and also refers to the corresponding amino acid sequence, for example, specified in SEQ ID NO:18, which corresponds to the protein sequence defined in the protein reference sequence under NCBI accession number NP_001184152 .1 encoding the PDE4D6 polypeptide.

The term "phosphodiesterase 4D8" or "PDE4D8" refers to splicing variant 8 of the human phosphodiesterase PDE4D, i.e. the human phosphodiesterase PDE4D8 gene, for example, the sequence defined in the NCBI reference sequence: NM_001197219.1, in particular the nucleotide sequence specified in SEQ ID NO:27, which corresponds to the sequence of the above NCBI reference sequence of the PDE4D8 transcript, and also refers to the corresponding amino acid sequence, for example, specified in SEQ ID NO:28, which corresponds to the protein sequence defined in the protein reference sequence under NCBI accession number NP_001184148 .1 encoding the PDE4D8 polypeptide.

The term "phosphodiesterase 4D9" or "PDE4D9" refers to splicing variant 9 of the human phosphodiesterase PDE4D, i.e. the human phosphodiesterase PDE4D9 gene, for example, the sequence defined in the NCBI reference sequence: NM_001197220.1, in particular the nucleotide sequence specified in SEQ ID NO:29, which corresponds to the sequence of the above NCBI reference sequence of the PDE4D9 transcript, and also refers to the corresponding amino acid sequence, for example, specified in SEQ ID NO:30, which corresponds to the protein sequence defined in the reference protein sequence under NCBI accession number NP_001184149 .1 encoding the PDE4D9 polypeptide. The term "phosphodiesterase 4D9" or "PDE4D9" also refers to an amplicon that can be generated using the primer pair PDE4D9_forward (SEQ ID NO:31) and PDE4D9_reverse (SEQ ID NO:32) and can be detected using the probe SEQ ID NO: 33.

The terms "PDE4D1", "PDE4D2", "PDE4D3", "PDE4D4", "PDE4D5", "PDE4D6", "PDE4D8", and "PDE4D9" also include nucleotide sequences showing a high degree of homology to PDE4D1, PDE4D2, PDE4D3, PDE4D4, PDE4D5, PDE4D6, PDE4D8, and PDE4D9, respectively, e.g., nucleic acid sequences that are at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96% , 97%, 98% or 99% identical to the sequence specified in SEQ ID NO:1, 6, 8, 10, 12, 17, 27 or 29, respectively, or amino acid sequences that are at least 75%, 80% , 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the sequence indicated in SEQ ID NO:2, 7, 9, 11, 13, 18, 28, or 30, respectively, or nucleic acid sequences encoding amino acid sequences that are at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95% , 96%, 97%, 98% or 99% identical to the sequence indicated in SEQ ID NO: 2, 7, 9, 11, 13, 18, 28 or 30, or amino acid sequences encoded by nucleic acid sequences that are at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence shown in SEQ ID NO:1, 6, 8, 10, 12, 17, 27 or 29.

Any reference positions in the claims are not to be construed as limiting the scope.

The present invention relates to a method for preoperative risk stratification in a subject suffering from prostate cancer, comprising determining the gene expression profile for phosphodiesterase 4D variant 7 (PDE4D7) in a biological sample obtained from the subject, determining the gene expression profile for DExH-box helicase 9 (DHX9 ) in the same or different biological sample obtained from the subject, and determining a preoperative predictive risk score for the subject based on the PDE4D7 gene expression profile and the DHX9 gene expression profile. This may provide improved stratification of the subject in the preoperative setting, which may lead to better decisions about primary treatment. For example, preoperative predictive risk assessment may provide better guidance on the choice of active surveillance versus active intervention, such as radical prostatectomy, for certain patient subpopulations with prostate cancer.

The attached sequence listing, entitled 2018PF00399_Sequence Listing_ST25, is incorporated herein by reference in its entirety.

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SEQUENCE LIST

<110> Koninklijke Philips N.V.

<120> PREOPERATIVE RISK STRATIFICATION BASED ON PDE4D7 AND DHX9 EXPRESSION

<130> 2018PF00399

<160> 75

<170>PatentIn version 3.5

<210> 1

<211> 7801

<212> DNA

<213> Homo sapiens

<400> 1

gtggtggccg cgcacccggc cgcggctgat tcattcactt caagtgccgt gcagaaggct 60

cggcaggcgg ggcgggcgtg gggccgcggc tccgggttgg ggaccgagga gatccggctg 120

tggaccagac gctcctctgc ggggcgggca cccaagcgcg ctcgccaccc cctcgccatc 180

cgctagagcc gggctcctgg actgggactc gggcccgccg cacagttgaa aagtcgcata 240

gtggtttttc cgctcgcgtc gctgtgtgaa agttggctcg ccgctctttg cacgccctcc 300

ctggaggccg acccgagacg ccaagctgga gagaccgtgc ctccccgagg ccggccgccc 360

cgcgagcaca gcctccgccc ccgttgcact gccgggctgg gcaatatgaa ggagcagccc 420

tcatgtgccg gcaccggggca tccgagcatg gcggggtatg gcaggatggc cccctttgaa 480

ctcgctagcg gacccgtgaa gcgcttgaga actgagtccc cctttccctg tctcttcgca 540

gaggaggcct accagaaact ggccagcgag accctggagg agctggactg gtgtctggac 600

cagctagaga ccctacagac caggcactcc gtcagtgaga tggcctccaa caagtttaaa 660

aggatgctta atcgggagct cacccatctc tctgaaatga gtcggtctgg aaatcaagtg 720

tcagagttta tatcaaacac attcttagat aagcaacatg aagtggaaat tccttctcca 780

actcagaagg aaaaggagaa aaagaaaaga ccaatgtctc agatcagtgg agtcaagaaa 840

ttgatgcaca gctctagtct gactaattca agtatcccaa ggtttggagt taaaactgaa 900

caagaagatg tccttgccaa ggaactagaa gatgtgaaca aatggggtct tcatgttttc 960

agaatagcag agttgtctgg taaccggccc ttgactgtta tcatgcacac catttttcag 1020

gaacgggatt tattaaaaac atttaaaatt ccagtagata ctttaattac atatcttatg 1080

actctcgaag accattacca tgctgatgtg gcctatcaca acaatatcca tgctgcagat 1140

1200

ttggagattc ttgcagcaat ttttgccagt gcaatacatg atgtagatca tcctggtgtg 1260

tccaatcaat ttctgatcaa tacaaactct gaacttgcct tgatgtacaa tgattcctca 1320

gtcttagaga accatcattt ggctgtgggc tttaaattgc ttcaggaaga aaactgtgac 1380

attttccaga atttgaccaa aaaacaaaga caatctttaa ggaaaatggt cattgacatc 1440

gtacttgcaa cagatatgtc aaaacacatg aatctactgg ctgatttgaa gactatggtt 1500

gaaactaaga aagtgacaag ctctggagtt cttcttcttg ataattattc cgataggatt 1560

caggttcttc agaatatggt gcactgtgca gatctgagca acccaacaaa gcctctccag 1620

ctgtaccgcc agtggacgga ccggataatg gaggagttct tccgccaagg agaccgagag 1680

agggaacgtg gcatggagat aagccccatg tgtgacaagc acaatgcttc cgtggaaaaa 1740

tcacaggtgg gcttcataga ctatattgtt catcccctct gggagacatg ggcagacctc 1800

gtccaccctg acgccgga tattttggac actttggagg acaatcgtga atggtaccag 1860

agcacaatcc ctcagagccc ctctcctgca cctgatgacc cagaggaggg ccggcagggt 1920

caaactgaga aattccagtt tgaactaact ttagaggaag atggtgagtc agacacggaa 1980

aaggacagtg gcagtcaagt ggaagaagac actagctgca gtgactccaa gactctttgt 2040

actcaagact cagagtctac tgaaattccc cttgatgaac aggttgaaga ggaggcagta 2100

ggggaagaag aggaaagcca gcctgaagcc tgtgtcatag atgatcgttc tcctgacacg 2160

2220

gcatgtcaca tgccacaacc acggtcacac ctcactgtca tctgccagga cgtttgttga 2280

acaaaactga ccttgactac tcagtccagc gctcaggaat atcgtaacca gttttttcac 2340

ctccatgtca tccgagcaag gtggacatct tcacgaacag cgtttttaac aagatttcag 2400

cttggtagag ctgacaaagc agataaaatc tactccaaat tattttcaag agagtgtgac 2460

2520

aatattttca gaagaaaggc attgcacaga gtgaacttaa tggacgaagc aacaaatatg 2580

tcaagaacag gacatagcac gaatctgtta ccagtaggag gaggatgagc cacagaaatt 2640

gcataatttt ctaatttcaa gtcttcctga tacatgactg aatagtgtgg ttcagtgagc 2700

2760

attattaaat gtattgaggt attatattta aaaaaaacta tgttcagaac ttcatctgcc 2820

actggttatt tttttctaag gagtaacttg caagttttca gtacaaatct gtgctacact 2880

ggataaaaat ctaatttatg aattttactt gcaccttata gttcatagca attaactgat 2940

ttgtagtgat tcattgtttg ttttatatac caatgacttc catattttaa aagagaaaaa 3000

3060

tcactctttc cagataagca gagttgctct tcaccagtgt ttttcttcat gtgcaaagtg 3120

actatttgtt ctataatact tttatgtgtg ttatatcaaa tgtgtcttaa gcttcatgca 3180

aactcagtca tcagttcgtg ttgtctgaag caagtgggag atatataaat acccagtagc 3240

taaaatggtc agtctttttt agatgttttc ctacttagta tctcctaata acgttttgct 3300

gtgtcactag atgttcattt cacaagtgca tgtctttcta ataatccaca catttcatgc 3360

3420

3480

ttgctagatg aggtcttgtc aaatatatca ctaccattgt caccggtgaa aagaaacagg 3540

3600

actcttggtt tacagagaaa agttaaacag ccaactaggc agtttttaag aatattaaca 3660

atatattaac aaacaccaat acaactaatc ctatttggtt ttaatgattt caccatggga 3720

ttaagaacta tatcaggaac atccctgaga aacggtttta agtgtagcaa ctactcttcc 3780

ttaatggaca gccacataac gtgtaggaag tcctttatca cttatcctcg atccataagc 3840

atatcttgca gaggggaact acttctttaa acacatggag ggaaagaaga tgatgccact 3900

ggcaccagag ggttagtact gtgatgcatc ctaaaatatt tattatattg gtaaaaattc 3960

tggttaaata aaaaattaga gatcactctt ggctgatttc agcaccagga actgtattac 4020

agttttagag attaattcct agtgtttacc tgattatagc agttggcatc atggggcatt 4080

taattctgac tttatcccca cgtcagcctt aataaagtct tctttacctt ctctatgaag 4140

actttaaagc ccaaataatc atttttcaca ttgatattca agaattgaga tagatagaag 4200

ccaaagtggg tatctgacaa gtggaaaatc aaacgtttaa gaagaattac aactctgaaa 4260

agcatttata tgtggaactt ctcaaggagc ctcctgggga ctggaaagta agtcatcagc 4320

caggcaaatg actcatgctg aagagagtcc ccatttcagt cccctgagat ctagctgatg 4380

cttagatcct ttgaaataaa aattatgtct ttataactct gatcttttac ataaagcaga 4440

agaggaatca actagttaat tgcaaggttt ctactctgtt tcctctgtaa agatcagatg 4500

gtaatctttc aaataagaaa aaaataaaga cgtatgtttg accaagtagt ttcacaagaa 4560

tatttgggaa cttgtttctt ttaattttat ttgtccctga gtgaagtcta gaaagaaagg 4620

taaagagtct agagtttatt cctctttcca aaacattctc attcctctcc tccctacact 4680

tagtatttcc ccccagagt gcctagaatc ttaataatga ataaaataaa aagcagcaat 4740

atgtcattaa caaatccaga cctgaaaggg taaagggttt ataactgcac taataaagag 4800

4860

cactaatgga caatcaaatt gcagaaaagg ctcaatatcc aagagacagg gactaatgca 4920

ctgtacaatc tgcttatcct tgcccttctc tcttgccaaa gtgtgcttca gaaatatata 4980

ctgctttaaa aaagaataaa agaatatcct tttacaagtg gctttacatt tcctaaaatg 5040

ccataagaaa atgcaatatc tgggtactgt atggggaaaa aaatgtccaa gtttgtgtaa 5100

5160

atatcagtta aaattcacaa taatgtagat agaacaaatt acagacaagg aaagaaaaaa 5220

5280

tttgtgccat gcattttttt tctcaccaaa tgaccttacc tgtaatacag tcttgtttgt 5340

ctgtttacaa ccatgtattt attgcaatgt acatactgta atgttaattg taaattatct 5400

gttcttatta aaacatcatc ccatgatggg atggtgttga tatatttggga aactcttggt 5460

gagagaatga atggtgtgta tacatactct gtacattttt cttttctcct gtaatatagt 5520

cttgtcacct tagagcttgt ttatggaaga ttcaagaaaa ctataaaata cttaaagata 5580

tataaattta aaaaaacata gctgcaggtc tttggtccca gggctgtgcc ttaactttaa 5640

ccaatatttt cttctgtttt gctgcatttg aaaggtaaca gtggagctag ggctgggcat 5700

tttacatcca ggcttttaat tgattagaat tctgccaata ggtggatttt acaaaaccac 5760

agacaacctc tgaaagattc tgagaccctt ttgagacaga agctcttaag tacttcttgc 5820

cagggagcag cactgcatgt gtgatggttg tttgccatct gttgatcagg aactacttca 5880

gctacttgca tttgattatt tccttttttt tttttttttaa ctcggaaaca caactgggga 5940

6000

tctagagctc ataggaaaat ggacttgatt tgaaattgga gccagagttt actcgtgttg 6060

gttatctatt catcagcttc ctgacatgtt aagagaatac attaaagaga aaatactgtt 6120

ttttaatcct aaaatttttc ttccactaag ataaaccaaa tgtccttaca tatatgtaaa 6180

catagcagaa agcattcact 6240

atcctctaag atttgtttct gcaaaacttt cattgcttta gaattttaaa atttcacctt 6300

gtacaatggc cagcccctaa agcaggaaac atttataatg gattatatgg aaacatcctc 6360

ccagtacttg cccagccctt gaatcatgtg gcttttcagt gaaaggaaag attctttttc 6420

6480

taaacagtac aaaggggaca 6540

ctataattac aaaaacatcc ttaactgatt tgagttgttt ttatttcttt ggatatattt 6600

tcagagtggt aaattgtgtg tgagaattac aaatgattat tcttttagtg gtttcttagc 6660

ctctcttaca gcccacgggg atagtactgt acatcaatac cttcatatga aatttttata 6720

tgcaatgaaa ataaaagcat gggttgattc tgcctattta tgactcaatc ttttacaaat 6780

aaaagattat tcattttaaa ttatagttca atcagcatgt ctcttaggat actgaacgtg 6840

gttgaaatga aaggatagtg acatcataag ttagtactga tattcataac caaataaagc 6900

cattaaaaat taccaaaatt acttagatgg cctataagat 6960

taagcatggt gttttctaag caagctttga aaggggcctt ccatacttac ttaattgaat 7020

attctgggat attgaaaatt attcagatac ttgacaatta ttttttggtta cctactccgc 7080

aaactacaaa gttttaagga ctcaacaata agttaatgag acacagtgtt tgctttcatg 7140

gagcttacag tctggagggg acaaaggctt aaacaatact catataatta tatatgtgat 7200

cagtacaatg aaggagctca gtggggtaaa taagcaggaa cctgaacttg atctgttccg 7260

gagggccaca gaaggcttcc ttgaggcctt gagaaagtga tttgcatctg agttctgaag 7320

7380

acatttgcaa agatcttgag gcataaatga gcttgagaca tctggagaaa ctgaggaaaa 7440

gtgagagagt aggcagggcc tggagccgca gagccattgc taaccatcct gtgtgagata 7500

tcccccattc tgtagcttta ttctcataac cctgctcaat tttctttata acacttctca 7560

cagatttata tacgtgtttg tttttgttat ctgtctctcc caccagacca cagctccatg 7620

agagcaaggt ctttgcttac caatatatca ctagcactta aaactatgcc tggtacacag 7680

taggttctta atatgtgttg aatatagcca tcaaattgat attggatata attcaatctg 7740

ataagatatt ttgagatatt aaagagtttt taacttgata ccataaaaaa aaaaaaaaaa 7800

a 7801

<210> 2

<211> 585

<212> PROTEIN

<213> Homo sapiens

<400> 2

Met Lys Glu Gln Pro Ser Cys Ala Gly Thr Gly His Pro Ser Met Ala

1 5 10 15

Gly Tyr Gly Arg Met Ala Pro Phe Glu Leu Ala Ser Gly Pro Val Lys

20 25 30

Arg Leu Arg Thr Glu Ser Pro Phe Pro Cys Leu Phe Ala Glu Glu Ala

35 40 45

Tyr Gln Lys Leu Ala Ser Glu Thr Leu Glu Glu Leu Asp Trp Cys Leu

50 55 60

Asp Gln Leu Glu Thr Leu Gln Thr Arg His Ser Val Ser Glu Met Ala

65 70 75 80

Ser Asn Lys Phe Lys Arg Met Leu Asn Arg Glu Leu Thr His Leu Ser

85 90 95

Glu Met Ser Arg Ser Gly Asn Gln Val Ser Glu Phe Ile Ser Asn Thr

100 105 110

Phe Leu Asp Lys Gln His Glu Val Glu Ile Pro Ser Pro Thr Gln Lys

115 120 125

Glu Lys Glu Lys Lys Lys Arg Pro Met Ser Gln Ile Ser Gly Val Lys

130 135 140

Lys Leu Met His Ser Ser Ser Leu Thr Asn Ser Ser Ile Pro Arg Phe

145 150 155 160

Gly Val Lys Thr Glu Gln Glu Asp Val Leu Ala Lys Glu Leu Glu Asp

165 170 175

Val Asn Lys Trp Gly Leu His Val Phe Arg Ile Ala Glu Leu Ser Gly

180 185 190

Asn Arg Pro Leu Thr Val Ile Met His Thr Ile Phe Gln Glu Arg Asp

195 200 205

Leu Leu Lys Thr Phe Lys Ile Pro Val Asp Thr Leu Ile Thr Tyr Leu

210 215 220

Met Thr Leu Glu Asp His Tyr His Ala Asp Val Ala Tyr His Asn Asn

225 230 235 240

Ile His Ala Ala Asp Val Val Gln Ser Thr His Val Leu Leu Ser Thr

245 250 255

Pro Ala Leu Glu Ala Val Phe Thr Asp Leu Glu Ile Leu Ala Ala Ile

260 265 270

Phe Ala Ser Ala Ile His Asp Val Asp His Pro Gly Val Ser Asn Gln

275 280 285

Phe Leu Ile Asn Thr Asn Ser Glu Leu Ala Leu Met Tyr Asn Asp Ser

290 295 300

Ser Val Leu Glu Asn His His Leu Ala Val Gly Phe Lys Leu Leu Gln

305 310 315 320

Glu Glu Asn Cys Asp Ile Phe Gln Asn Leu Thr Lys Lys Gln Arg Gln

325 330 335

Ser Leu Arg Lys Met Val Ile Asp Ile Val Leu Ala Thr Asp Met Ser

340 345 350

Lys His Met Asn Leu Leu Ala Asp Leu Lys Thr Met Val Glu Thr Lys

355 360 365

Lys Val Thr Ser Ser Gly Val Leu Leu Leu Asp Asn Tyr Ser Asp Arg

370 375 380

Ile Gln Val Leu Gln Asn Met Val His Cys Ala Asp Leu Ser Asn Pro

385 390 395 400

Thr Lys Pro Leu Gln Leu Tyr Arg Gln Trp Thr Asp Arg Ile Met Glu

405 410 415

Glu Phe Phe Arg Gln Gly Asp Arg Glu Arg Glu Arg Gly Met Glu Ile

420 425 430

Ser Pro Met Cys Asp Lys His Asn Ala Ser Val Glu Lys Ser Gln Val

435 440 445

Gly Phe Ile Asp Tyr Ile Val His Pro Leu Trp Glu Thr Trp Ala Asp

450 455 460

Leu Val His Pro Asp Ala Gln Asp Ile Leu Asp Thr Leu Glu Asp Asn

465 470 475 480

Arg Glu Trp Tyr Gln Ser Thr Ile Pro Gln Ser Pro Ser Pro Ala Pro

485 490 495

Asp Asp Pro Glu Glu Gly Arg Gln Gly Gln Thr Glu Lys Phe Gln Phe

500 505 510

Glu Leu Thr Leu Glu Glu Asp Gly Glu Ser Asp Thr Glu Lys Asp Ser

515 520 525

Gly Ser Gln Val Glu Glu Asp Thr Ser Cys Ser Asp Ser Lys Thr Leu

530 535 540

Cys Thr Gln Asp Ser Glu Ser Thr Glu Ile Pro Leu Asp Glu Gln Val

545 550 555 560

Glu Glu Glu Ala Val Gly Glu Glu Glu Glu Ser Gln Pro Glu Ala Cys

565 570 575

Val Ile Asp Asp Arg Ser Pro Asp Thr

580 585

<210> 3

<211> 21

<212> DNA

<213> Artificial sequence

<220>

<223> PDE4D1D2_forward primer

<400> 3

aatatgaagg agcagccctc a 21

<210> 4

<211> 18

<212> DNA

<213> Artificial sequence

<220>

<223> PDE4D1D2_reverse primer

<400> 4

gtctcgctgg ccagtttc 18

<210> 5

<211> 18

<212> DNA

<213> Artificial sequence

<220>

<223> PDE4D1D2 probe

<400> 5

catccgagca tggcggga 18

<210> 6

<211> 7715

<212> DNA

<213> Homo sapiens

<400> 6

gtggtggccg cgcacccggc cgcggctgat tcattcactt caagtgccgt gcagaaggct 60

cggcaggcgg ggcgggcgtg gggccgcggc tccgggttgg ggaccgagga gatccggctg 120

tggaccagac gctcctctgc ggggcgggca cccaagcgcg ctcgccaccc cctcgccatc 180

cgctagagcc gggctcctgg actgggactc gggcccgccg cacagttgaa aagtcgcata 240

gtggtttttc cgctcgcgtc gctgtgtgaa agttggctcg ccgctctttg cacgccctcc 300

ctggaggccg acccgagacg ccaagctgga gagaccgtgc ctccccgagg ccggccgccc 360

cgcgagcaca gcctccgccc ccgttgcact gccgggctgg gcaatatgaa ggagcagccc 420

tcatgtgccg gcaccggggca tccgagcatg gcgggaggag gcctaccaga aactggccag 480

cgagaccctg gaggagctgg actggtgtct ggaccagcta gagaccctac agaccaggca 540

ctccgtcagt gagatggcct ccaacaagtt taaaaggatg cttaatcggg agctcaccca 600

tctctctgaa atgagtcggt ctggaaatca agtgtcagag tttatatcaa acacattctt 660

agataagcaa catgaagtgg aaattccttc tccaactcag aaggaaaagg agaaaaagaa 720

aagaccaatg tctcagatca gtggagtcaa gaaattgatg cacagctcta gtctgactaa 780

ttcaagtatc ccaaggtttg gagttaaaac tgaacaagaa gatgtccttg ccaaggaact 840

agaagatgtg aacaaatggg gtcttcatgt tttcagaata gcagagttgt ctggtaaccg 900

gcccttgact gttatcatgc acaccatttt tcaggaacgg gatttattaa aaacatttaa 960

aattccagta gatactttaa ttacatatct tatgactctc gaagaccatt accatgctga 1020

tgtggcctat cacaacaata tccatgctgc agatgttgtc cagtctactc atgtgctatt 1080

1140

cagtgcaata catgatgtag atcatcctgg tgtgtccaat caatttctga tcaatacaaa 1200

ctctgaactt gccttgatgt acaatgattc ctcagtctta gagaaccatc atttggctgt 1260

gggctttaaa ttgcttcagg aagaaaactg tgacattttc cagaatttga ccaaaaaaca 1320

1380

catgaatcta ctggctgatt tgaagactat ggttgaaact aagaaagtga caagctctgg 1440

agttcttctt cttgataatt attccgatag gattcaggtt cttcagaata tggtgcactg 1500

tgcagatctg agcaacccaa caaagcctct ccagctgtac cgccagtgga cggaccggat 1560

aatggaggag ttcttccgcc aaggagaccg agagagggaa cgtggcatgg agataagccc 1620

catgtgtgac aagcacaatg cttccgtgga aaaatcacag gtgggcttca tagactatat 1680

tgttcatccc ctctgggaga catgggcaga cctcgtccac cctgacgccc aggatatttt 1740

ggacactttg gaggacaatc gtgaatggta ccagagcaca atccctcaga gcccctctcc 1800

tgcacctgat gacccagagg agggccggca gggtcaaact gagaaattcc agtttgaact 1860

aactttagag gaagatggtg agtcagacac ggaaaaggac agtggcagtc aagtggaaga 1920

agacactagc tgcagtgact ccaagactct ttgtactcaa gactcagagt ctactgaaat 1980

2040

agcctgtgtc atagatgatc gttctcctga cacgtaacag tgcaaaaact ttcatgcctt 2100

tttttttttt aagtagaaaa attgtttcca aagtgcatgt cacatgccac aaccacggtc 2160

acacctcact gtcatctgcc aggacgtttg ttgaacaaaa ctgaccttga ctactcagtc 2220

cagcgctcag gaatatcgta accagttttt tcacctccat gtcatccgag caaggtggac 2280

2340

aatctactcc aaattatttt caagagagtg tgactcatca ggcagcccaa aagtttattg 2400

gacttggggt ttctattcct ttttatttgt ttgcaatatt ttcagaagaa aggcattgca 2460

cagagtgaac ttaatggacg aagcaacaaa tatgtcaaga acaggacata gcacgaatct 2520

gttaccagta ggaggaggat gagccacaga aattgcataa ttttctaatt tcaagtcttc 2580

ctgatacatg actgaatagt gtggttcagt gagctgcact gacctctaca ttttgtatga 2640

tatgtaaaac agattttttg tagagcttac ttttattatt aaatgtattg aggtattata 2700

tttaaaaaaa actatgttca gaacttcatc tgccactggt tatttttttc taaggagtaa 2760

cttgcaagtt ttcagtacaa atctgtgcta cactggataa aaatctaatt tatgaatttt 2820

acttgcacct tatagttcat agcaattaac tgatttgtag tgattcattg tttgttttat 2880

ataccaatga cttccatatt ttaaaagaga aaaacaactt tatgttgcag gaaacccttt 2940

ttgtaagtct ttattattta ctttgcattt tgtttcactc tttccagata agcagagttg 3000

3060

3120

aaatacccag tagctaaaat ggtcagtctt ttttagatgt 3180

tttcctactt agtatctcct aataacgttt tgctgtgtca ctagatgttc atttcacaag 3240

tgcatgtctt tctaataatc cacacatttc atgctctaat aatccacaca tttcatgctc 3300

atttttattg tttttacagc cagttatagt aagaaaaagg tttttcccct tgtgctgctt 3360

tataatttag cgtgtgtctg aaccttatcc atgttgcta gatgaggtct tgtcaaatat 3420

atcactacca ttgtcaccgg tgaaaagaaa caggtagtta agttagggtt aacattcatt 3480

tcaaccacga ggttgtatat catgactagc ttttactctt ggtttacaga gaaaagttaa 3540

acagccaact aggcagtttt taagaatatt aacaatatat taacaaacac caatacaact 3600

3660

gagaaacggt tttaagtgta gcaactactc ttccttaatg gacagccaca taacgtgtag 3720

gaagtccttt atcacttatc ctcgatccat aagcatatct tgcagagggg aactacttct 3780

ttaaacacat ggagggaaag aagatgatgc cactggcacc aggggttag tactgtgatg 3840

catcctaaaa tatttattat attggtaaaa attctggtta aataaaaaat tagagatcac 3900

3960

tacctgatta tagcagttgg catcatgggg catttaattc tgactttatc cccacgtcag 4020

ccttaataaa gtcttcttta ccttctctat gaagacttta aagcccaaat aatcattttt 4080

cacattgata ttcaagaatt gagatagata gaagccaaag tgggtatctg acaagtggaa 4140

aatcaaacgt ttaagaagaa ttacaactct gaaaagcatt tatatgtgga acttctcaag 4200

gagcctcctg gggactggaa agtaagtcat cagccaggca aatgactcat gctgaagaga 4260

gtccccattt cagtcccctg agatctagct gatgcttaga tcctttgaaa taaaaattat 4320

gtctttataa ctctgatctt ttacataaag cagagagga atcaactagt taattgcaag 4380

gtttctactc tgtttcctct gtaaagatca gatggtaatc tttcaaataa gaaaaaaata 4440

aagacgtatg tttgaccaag tagtttcaca agaatatttg ggaacttgtt tcttttaatt 4500

ttatttgtcc ctgagtgaag tctagaaaga aaggtaaaga gtctagagtt tattcctctt 4560

tccaaaacat tctcattcct ctcctcccta cacttagtat ttcccccaca gagtgcctag 4620

aatcttaata atgaataaaa taaaaagcag caatatgtca ttaacaaatc cagacctgaa 4680

agggtaaagg gtttataact gcactaataa agagaggctc ttttttttttc ttccagtttg 4740

ttggttttta atggtaccgt gttgtaaaga tacccactaa tggacaatca aattgcagaa 4800

aaggctcaat atccaagaga cagggactaa tgcactgtac aatctgctta tccttgccct 4860

tctctcttgc caaagtgtgc ttcagaaata tatactgctt taaaaaagaa taaaagaata 4920

tccttttaca agtggcttta catttcctaa aatgccataa gaaaatgcaa tatctgggta 4980

ctgtatgggg aaaaaaatgt ccaagtttgt gtaaaaccag tgcatttcag cttgcaagtt 5040

5100

agatagaaca aattacagac aaggaaagaa aaaacttgaa tgaaatggat tttacagaaa 5160

gctttatgat aatttttgaa tgcattattt attttttgtg ccatgcattt tttttctcac 5220

caaatgacct tacctgtaat acagtcttgt ttgtctgttt acaaccatgt atttattgca 5280

atgtacatac tgtaatgtta attgtaaatt atctgttctt attaaaacat catcccatga 5340

5400

ctctgtacat ttttcttttc tcctgtaata tagtcttgtc accttagagc ttgtttatgg 5460

aagattcaag aaaactataa aatacttaaa gatatataaa tttaaaaaaa catagctgca 5520

ggtctttggt cccagggctg tgccttaact ttaaccaata ttttcttctg ttttgctgca 5580

tttgaaaggt aacagtggag ctagggctgg gcattttaca tccaggcttt taattgatta 5640

gaattctgcc aataggtgga ttttacaaaa ccacagacaa cctctgaaag attctgagac 5700

ccttttgaga cagaagctct taagtacttc ttgccaggga gcagcactgc atgtgtgatg 5760

gttgtttgcc atctgttgat caggaactac ttcagctact tgcatttgat tatttccttt 5820

tttttttttt ttaactcgga aacacaactg gggaaatata ttctttccca gtgattataa 5880

acaatctttt tctttttttt aagtcctttt ggcttctaga gctcatagga aaatggactt 5940

gatttgaaat tggagccaga gtttactcgt gttggttatc tattcatcag cttcctgaca 6000

tgttaagaga atacattaaa gagaaaatac

6120

6180

ctttcattgc tttgaattt taaaatttca ccttgtacaa tggccagccc ctaaagcagg 6240

aaacatttat aatggattat atggaaacat cctcccagta cttgcccagc ccttgaatca 6300

6360

attttatttt attttttgac acaaactgta gattttagca gccctggccc aaaggaattt 6420

gattactttt gttttaaaca gtacaaaggg gacactataa ttacaaaaac atccttaact 6480

6540

ttacaaatga ttattctttt agtggtttct tagcctctct tacagcccac ggggatagta 6600

ctgtacatca ataccttcat atgaaatttt tatatgcaat gaaaataaaa gcatgggttg 6660

6720

atgtctctta ggatactgaa cgtggttgaa atgaaaggat agtgacatca 6780

taagttagta ctgatattca taaccaaata aagccaactt gagtaatttt gctacattaa 6840

aaattaccaa aattacttag atggcctata agattaagca tggtgttttc taagcaagct 6900

ttgaaagggg ccttccatac ttacttaatt gaatattctg ggatattgaa aattattcag 6960

atacttgaca attatttttg gttacctact ccgcaaacta caaagtttta aggactcaac 7020

aataagttaa tgagacacag tgtttgcttt catggagctt acagtctgga ggggacaaag 7080

gcttaaacaa tactcatata attatatatg tgatcagtac aatgaaggag ctcagtgggg 7140

taaataagca ggaacctgaa cttgatctgt tccggagggc cacagaaggc ttccttgagg 7200

ccttgagaaa gtgatttgca tctgagttct gaaggattgt aagaggtaac tagggaaaaa 7260

gttgacagga agaggaaggg gatccagaca agaaacattt gcaaagatct tgaggcataa 7320

atgagcttga gacatctgga gaaactgagg aaaagtgaga gagtaggcag ggcctggagc 7380

cgcagagcca ttgctaacca tcctgtgtga gatatccccc attctgtagc tttattctca 7440

taaccctgct caattttctt tataacactt ctcacagatt tatatacgtg tttgtttttg 7500

ttatctgtct ctcccaccag accacagctc catgagagca aggtctttgc ttaccaatat 7560

atcactagca cttaaaacta tgcctggtac acagtaggtt cttaatatgt gttgaatata 7620

7680

tttttaactt gataccataa aaaaaaaaaa aaaaa 7715

<210> 7

<211> 507

<212> PROTEIN

<213> Homo sapiens

<400> 7

Met Ala Ser Asn Lys Phe Lys Arg Met Leu Asn Arg Glu Leu Thr His

1 5 10 15

Leu Ser Glu Met Ser Arg Ser Gly Asn Gln Val Ser Glu Phe Ile Ser

20 25 30

Asn Thr Phe Leu Asp Lys Gln His Glu Val Glu Ile Pro Ser Pro Thr

35 40 45

Gln Lys Glu Lys Glu Lys Lys Lys Arg Pro Met Ser Gln Ile Ser Gly

50 55 60

Val Lys Lys Leu Met His Ser Ser Ser Leu Thr Asn Ser Ser Ile Pro

65 70 75 80

Arg Phe Gly Val Lys Thr Glu Gln Glu Asp Val Leu Ala Lys Glu Leu

85 90 95

Glu Asp Val Asn Lys Trp Gly Leu His Val Phe Arg Ile Ala Glu Leu

100 105 110

Ser Gly Asn Arg Pro Leu Thr Val Ile Met His Thr Ile Phe Gln Glu

115 120 125

Arg Asp Leu Leu Lys Thr Phe Lys Ile Pro Val Asp Thr Leu Ile Thr

130 135 140

Tyr Leu Met Thr Leu Glu Asp His Tyr His Ala Asp Val Ala Tyr His

145 150 155 160

Asn Asn Ile His Ala Ala Asp Val Val Gln Ser Thr His Val Leu Leu

165 170 175

Ser Thr Pro Ala Leu Glu Ala Val Phe Thr Asp Leu Glu Ile Leu Ala

180 185 190

Ala Ile Phe Ala Ser Ala Ile His Asp Val Asp His Pro Gly Val Ser

195 200 205

Asn Gln Phe Leu Ile Asn Thr Asn Ser Glu Leu Ala Leu Met Tyr Asn

210 215 220

Asp Ser Ser Val Leu Glu Asn His His Leu Ala Val Gly Phe Lys Leu

225 230 235 240

Leu Gln Glu Glu Asn Cys Asp Ile Phe Gln Asn Leu Thr Lys Lys Gln

245 250 255

Arg Gln Ser Leu Arg Lys Met Val Ile Asp Ile Val Leu Ala Thr Asp

260 265 270

Met Ser Lys His Met Asn Leu Leu Ala Asp Leu Lys Thr Met Val Glu

275 280 285

Thr Lys Lys Val Thr Ser Ser Gly Val Leu Leu Leu Asp Asn Tyr Ser

290 295 300

Asp Arg Ile Gln Val Leu Gln Asn Met Val His Cys Ala Asp Leu Ser

305 310 315 320

Asn Pro Thr Lys Pro Leu Gln Leu Tyr Arg Gln Trp Thr Asp Arg Ile

325 330 335

Met Glu Glu Phe Phe Arg Gln Gly Asp Arg Glu Arg Glu Arg Gly Met

340 345 350

Glu Ile Ser Pro Met Cys Asp Lys His Asn Ala Ser Val Glu Lys Ser

355 360 365

Gln Val Gly Phe Ile Asp Tyr Ile Val His Pro Leu Trp Glu Thr Trp

370 375 380

Ala Asp Leu Val His Pro Asp Ala Gln Asp Ile Leu Asp Thr Leu Glu

385 390 395 400

Asp Asn Arg Glu Trp Tyr Gln Ser Thr Ile Pro Gln Ser Pro Ser Pro

405 410 415

Ala Pro Asp Asp Pro Glu Glu Gly Arg Gln Gly Gln Thr Glu Lys Phe

420 425 430

Gln Phe Glu Leu Thr Leu Glu Glu Asp Gly Glu Ser Asp Thr Glu Lys

435 440 445

Asp Ser Gly Ser Gln Val Glu Glu Asp Thr Ser Cys Ser Asp Ser Lys

450 455 460

Thr Leu Cys Thr Gln Asp Ser Glu Ser Thr Glu Ile Pro Leu Asp Glu

465 470 475 480

Gln Val Glu Glu Glu Glu Ala Val Gly Glu Glu Glu Glu Ser Gln Pro Glu

485 490 495

Ala Cys Val Ile Asp Asp Arg Ser Pro Asp Thr

500 505

<210> 8

<211> 7783

<212> DNA

<213> Homo sapiens

<400> 8

aatacttgtt gcaataattg cccacgatag ctgctcaaac aagagagttg gaattcatct 60

gtaaaaatca ctacatgtaa cgtaggagac aagaaaaata ttaatgacag aagatctgcg 120

aacatgatgc acgtgaataa ttttcccttt agaaggcatt cctggatatg ttttgatgtg 180

gacaatggca catctgcggg acggagtccc ttggatccca tgaccagccc aggaccgggg 240

ctaattctcc aagcaaattt tgtccacagt caacgacggg agtccttcct gtatcgatcc 300

gacagcgatt atgacctctc tccaaagtct atgtccggga actcctccat tgccagtgat 360

atacacggag atgacttgat tgtgactcca tttgctcagg tcttggccag tctgcgaact 420

gtacgaaaca actttgctgc attaactaat ttgcaagatc gagcacctag caaaagatca 480

cccatgtgca accaaccatc catcaacaaa gccaccataa cagagggaggc ctaccagaaa 540

ctggccagcg agaccctgga ggagctggac tggtgtctgg accagctaga gaccctacag 600

accaggcact ccgtcagtga gatggcctcc aacaagttta aaaggatgct taatcgggag 660

ctcacccatc tctctgaaat gagtcggtct ggaaatcaag tgtcagagtt tatatcaaac 720

acattcttag ataagcaaca tgaagtggaa attccttctc caactcagaa ggaaaaggag 780

aaaaagaaaa gaccaatgtc tcagatcagt ggagtcaaga aattgatgca cagctctagt 840

ctgactaatt caagtatccc aaggtttgga gttaaaactg aacaagaaga tgtccttgcc 900

aaggaactag aagatgtgaa caaatggggt cttcatgttt tcagaatagc agagttgtct 960

ggtaaccggc ccttgactgt tatcatgcac accatttttc aggaacggga tttattaaaa 1020

acatttaaaa ttccagtaga tactttaatt acatatctta tgactctcga agaccattac 1080

catgctgatg tggcctatca caacaatatc catgctgcag atgttgtcca gtctactcat 1140

1200

atttttgcca gtgcaataca tgatgtagat catcctggtg tgtccaatca atttctgatc 1260

aatacaaact ctgaacttgc cttgatgtac aatgattcct cagtcttaga gaaccatcat 1320

ttggctgtgg gctttaaatt gcttcaggaa gaaaactgtg acattttcca gaatttgacc 1380

aaaaaacaaa gacaatcttt aaggaaaatg gtcattgaca tcgtacttgc aacagatatg 1440

tcaaaacaca tgaatctact ggctgatttg aagactatgg ttgaaactaa gaaagtgaca 1500

agctctggag ttcttcttct tgataattat tccgatagga ttcaggttct tcagaatatg 1560

gtgcactgtg cagatctgag caacccaaca aagcctctcc agctgtaccg ccagtggacg 1620

gaccggataa tggaggagtt cttccgccaa ggagaccgag agagggaacg tggcatggag 1680

ataagcccca tgtgtgacaa gcacaatgct tccgtggaaa aatcacaggt gggcttcata 1740

1800

gatattttgg acactttgga ggacaatcgt gaatggtacc agagcacaat ccctcagagc 1860

ccctctcctg cacctgatga cccagaggag ggccggcagg gtcaaactga gaaattccag 1920

tttgaactaa ctttagagga agatggtgag tcagacacgg aaaaggacag tggcagtcaa 1980

gtggaagaag acactagctg cagtgactcc aagactcttt gtactcaaga ctcagagtct 2040

actgaaattc cccttgatga acaggttgaa gaggaggcag taggggaaga agaggaaagc 2100

2160

catgcctttt ttttttttaa gtagaaaaat tgtttccaaa gtgcatgtca catgccacaa 2220

ccacggtcac acctcactgt catctgccag gacgtttgtt gaacaaaact gaccttgact 2280

actcagtcca gcgctcagga atatcgtaac cagttttttc acctccatgt catccgagca 2340

aggtggacat cttcacgaac agcgttttta acaagatttc agcttggtag agctgacaaa 2400

gcagataaaa tctactccaa attattttca agagagtgtg actcatcagg cagcccaaaa 2460

gtttattgga cttggggttt ctattccttt ttatttgttt gcaatatttt cagaagaaag 2520

gcattgcaca gagtgaactt aatggacgaa gcaacaaata tgtcaagaac aggacatagc 2580

acgaatctgt taccagtagg aggaggatga gccacagaaa ttgcataatt ttctaatttc 2640

aagtcttcct gatacatgac tgaatagtgt ggttcagtga gctgcactga cctctacatt 2700

ttgtatgata tgtaaaacag attttttgta gagcttactt ttatttattaa atgtattgag 2760

gtattatatt taaaaaaaac tatgttcaga acttcatctg ccactggtta tttttttcta 2820

aggagtaact tgcaagtttt cagtacaaat ctgtgctaca ctggataaaa atctaattta 2880

tgaattttac ttgcacctta tagttcatag caattaactg atttgtagtg attcattgtt 2940

tgttttatat accaatgact tccatatttt aaaagagaaa aacaacttta tgttgcagga 3000

3060

cagagttgct cttcaccagt gtttttcttc atgtgcaaag tgactatttg ttctataata 3120

3180

3240

ttagatgttt tcctacttag tatctcctaa taacgttttg ctgtgtcact agatgttcat 3300

ttcacaagtg catgtctttc taataatcca cacatttcat gctctaataa tccacacatt 3360

3420

3480

tcaaatatat cactaccatt gtcaccggtg aaaagaaaca ggtagttaag ttagggttaa 3540

cattcatttc aaccacgagg ttgtatatca tgactagctt ttactcttgg tttacagaga 3600

aaagttaaac agccaactag gcagttttta agaatattaa caatatatta acaaacacca 3660

atacaactaa tcctatttgg ttttaatgat ttcaccatgg

acatccctga gaaacggttt taagtgtagc aactactctt ccttaatgga cagccacata 3780

acgtgtagga agtcctttat cacttatcct cgatccataa gcatatcttg cagaggggaa 3840

ctacttcttt aaacacatgg agggaaagaa gatgatgcca ctggcaccag agggttagta 3900

3960

gagatcactc ttggctgatt tcagcaccag gaactgtatt acagttttag agattaattc 4020

ctagtgttta cctgattata gcagttggca tcatggggca tttaattctg actttatccc 4080

cacgtcagcc ttaataaagt cttctttacc ttctctatga agactttaaa gcccaaataa 4140

tcatttttca cattgatatt caagaattga gatagataga agccaaagtg ggtatctgac 4200

aagtggaaaa tcaaacgttt aagaagaatt acaactctga aaagcattta tatgtggaac 4260

ttctcaagga gcctcctggg gactggaaag taagtcatca gccaggcaaa tgactcatgc 4320

tgaagagagt ccccatttca gtcccctgag atctagctga tgcttagatc ctttgaaata 4380

aaaattatgt ctttataact ctgatctttt acataaagca gaagaggaat caactagtta 4440

attgcaaggt ttctactctg tttcctctgt aaagatcaga tggtaatctt tcaaataaga 4500

aaaaaataaa gacgtatgtt tgaccaagta gtttcacaag aatatttggg aacttgtttc 4560

ttttaatttt atttgtccct gagtgaagtc tagaaagaaa ggtaaagagt ctagagttta 4620

ttcctctttc caaaacattc tcattcctct cctccctaca cttagtattt cccccacaga 4680

gtgcctagaa tcttaataat gaataaaata aaaagcagca atatgtcatt aacaaatcca 4740

gacctgaaag ggtaaagggt ttataactgc actaataaag agaggctctt tttttttctt 4800

ccagtttgtt ggtttttaat ggtaccgtgt tgtaaagata cccactaatg gacaatcaaa 4860

ttgcagaaaa ggctcaatat ccaagagaca gggactaatg cactgtacaa tctgcttatc 4920

cttgcccttc tctcttgcca aagtgtgctt cagaaatata tactgcttta aaaaagaata 4980

5040

tctgggtact gtatggggaa aaaaatgtcc aagtttgtgt aaaaccagtg catttcagct 5100

5160

aataatgtag atagaacaaa ttacagacaa ggaaagaaaa aacttgaatg aaatggattt 5220

tacagaaagc tttatgataa tttttgaatg cattattat tttttgtgcc atgcattttt 5280

tttctcacca aatgacctta cctgtaatac agtcttgttt gtctgtttac aaccatgtat 5340

ttattgcaat gtacatactg taatgttaat tgtaaattat ctgttcttat taaaacatca 5400

tcccatgatg ggatggtgtt gatatatttg gaaactcttg gtgagagaat gaatggtgtg 5460

tatacatact ctgtacattt ttcttttctc ctgtaatata gtcttgtcac cttagagctt 5520

gtttatggaa gattcaagaa aactataaaa tacttaaaga tatataaatt taaaaaaaca 5580

tagctgcagg tctttggtcc cagggctgtg ccttaacttt aaccaatatt ttcttctgtt 5640

ttgctgcatt tgaaaggtaa cagtggagct agggctgggc attttacatc caggctttta 5700

attgattaga attctgccaa taggtggatt ttacaaaacc acagacaacc tctgaaagat 5760

tctgagaccc ttttgagaca gaagctctta agtacttctt gccagggagc agcactgcat 5820

gtgtgatggt tgtttgccat ctgttgatca ggaactactt cagctacttg catttgatta 5880

ttttccttttt tttttttttt aactcggaaa cacaactggg gaaatatatt ctttcccagt 5940

gattataaac aatctttttc tttttttttaa gtccttttgg cttctagagc tcataggaaa 6000

6060

6120

tcttcacta agataaacca 6180

aaggtgggtt gatgtcagtt tacatagcag aaagcattca ctatcctcta agatttgttt 6240

ctgcaaaact ttcattgctt tagaatttta aaatttcacc ttgtacaatg gccagcccct 6300

6360

ttgaatcatg tggcttttca gtgaaaggaa agattctttt tctaggaaaa atgagcctat 6420

tttattttat tttattttat tttttgacac aaactgtaga ttttagcagc cctggcccaa 6480

aggaatttga ttacttttgt tttaaacagt acaaagggga cactataatt acaaaaacat 6540

ccttaactga tttgagttgt ttttatttct ttggatatat tttcagagtg gtaaattgtg 6600

tgtgagaatt acaaatgatt attcttttag tggtttctta gcctctctta cagcccacgg 6660

ggatagtact gtacatcaat accttcatat gaaattttta tatgcaatga aaataaaagc 6720

6780

aattatagtt caatcagcat gtctcttagg atactgaacg tggttgaaat gaaaggatag 6840

tgacatcata agttagtact gatattcata accaaataaa gccaacttga gtaattttgc 6900

tacattaaaa attaccaaaa ttacttagat ggcctataag attaagcatg gtgttttcta 6960

agcaagcttt gaaaggggcc ttccatactt acttaattga atattctggg atattgaaaa 7020

tattcagat acttgacaat tatttttggt tacctactcc gcaaactaca aagttttaag 7080

7140

ggacaaaggc ttaaacaata ctcatataat tatatatgtg atcagtacaa tgaaggagct 7200

cagtggggta aataagcagg aacctgaact tgatctgttc cggagggcca cagaaggctt 7260

ccttgaggcc ttgagaaagt gatttgcatc tgagttctga aggattgtaa gaggtaacta 7320

gggaaaaagt tgacaggaag aggaagggga tccagacaag aaacatttgc aaagatcttg 7380

aggcataaat gagcttgaga catctggaga aactgaggaa aagtgagaga gtaggcaggg 7440

cctggagccg cagagccatt gctaaccatc ctgtgtgaga tatcccccat tctgtagctt 7500

tattctcata accctgctca attttcttta taacacttct cacagattta tatacgtgtt 7560

tgtttttgtt atctgtctct cccaccagac cacagctcca tgagagcaag gtctttgctt 7620

accaatatat cactagcact taaaactatg cctggtacac agtaggttct taatatgtgt 7680

tgaatatagc catcaaattg atattggata taattcaatc tgataagata ttttgagata 7740

ttaaagagtt tttaacttga taccataaaa aaaaaaaaaa aaa 7783

<210> 9

<211> 673

<212> PROTEIN

<213> Homo sapiens

<400> 9

Met Met His Val Asn Asn Phe Pro Phe Arg Arg His Ser Trp Ile Cys

1 5 10 15

Phe Asp Val Asp Asn Gly Thr Ser Ala Gly Arg Ser Pro Leu Asp Pro

20 25 30

Met Thr Ser Pro Gly Ser Gly Leu Ile Leu Gln Ala Asn Phe Val His

35 40 45

Ser Gln Arg Arg Glu Ser Phe Leu Tyr Arg Ser Asp Ser Asp Tyr Asp

50 55 60

Leu Ser Pro Lys Ser Met Ser Arg Asn Ser Ser Ile Ala Ser Asp Ile

65 70 75 80

His Gly Asp Asp Leu Ile Val Thr Pro Phe Ala Gln Val Leu Ala Ser

85 90 95

Leu Arg Thr Val Arg Asn Asn Phe Ala Ala Leu Thr Asn Leu Gln Asp

100 105 110

Arg Ala Pro Ser Lys Arg Ser Pro Met Cys Asn Gln Pro Ser Ile Asn

115 120 125

Lys Ala Thr Ile Thr Glu Glu Ala Tyr Gln Lys Leu Ala Ser Glu Thr

130 135 140

Leu Glu Glu Leu Asp Trp Cys Leu Asp Gln Leu Glu Thr Leu Gln Thr

145 150 155 160

Arg His Ser Val Ser Glu Met Ala Ser Asn Lys Phe Lys Arg Met Leu

165 170 175

Asn Arg Glu Leu Thr His Leu Ser Glu Met Ser Arg Ser Gly Asn Gln

180 185 190

Val Ser Glu Phe Ile Ser Asn Thr Phe Leu Asp Lys Gln His Glu Val

195 200 205

Glu Ile Pro Ser Pro Thr Gln Lys Glu Lys Glu Lys Lys Lys Arg Pro

210 215 220

Met Ser Gln Ile Ser Gly Val Lys Lys Leu Met His Ser Ser Ser Leu

225 230 235 240

Thr Asn Ser Ser Ile Pro Arg Phe Gly Val Lys Thr Glu Gln Glu Asp

245 250 255

Val Leu Ala Lys Glu Leu Glu Asp Val Asn Lys Trp Gly Leu His Val

260 265 270

Phe Arg Ile Ala Glu Leu Ser Gly Asn Arg Pro Leu Thr Val Ile Met

275 280 285

His Thr Ile Phe Gln Glu Arg Asp Leu Leu Lys Thr Phe Lys Ile Pro

290 295 300

Val Asp Thr Leu Ile Thr Tyr Leu Met Thr Leu Glu Asp His Tyr His

305 310 315 320

Ala Asp Val Ala Tyr His Asn Asn Ile His Ala Ala Asp Val Val Gln

325 330 335

Ser Thr His Val Leu Leu Ser Thr Pro Ala Leu Glu Ala Val Phe Thr

340 345 350

Asp Leu Glu Ile Leu Ala Ala Ile Phe Ala Ser Ala Ile His Asp Val

355 360 365

Asp His Pro Gly Val Ser Asn Gln Phe Leu Ile Asn Thr Asn Ser Glu

370 375 380

Leu Ala Leu Met Tyr Asn Asp Ser Ser Val Leu Glu Asn His His Leu

385 390 395 400

Ala Val Gly Phe Lys Leu Leu Gln Glu Glu Asn Cys Asp Ile Phe Gln

405 410 415

Asn Leu Thr Lys Lys Gln Arg Gln Ser Leu Arg Lys Met Val Ile Asp

420 425 430

Ile Val Leu Ala Thr Asp Met Ser Lys His Met Asn Leu Leu Ala Asp

435 440 445

Leu Lys Thr Met Val Glu Thr Lys Lys Val Thr Ser Ser Gly Val Leu

450 455 460

Leu Leu Asp Asn Tyr Ser Asp Arg Ile Gln Val Leu Gln Asn Met Val

465 470 475 480

His Cys Ala Asp Leu Ser Asn Pro Thr Lys Pro Leu Gln Leu Tyr Arg

485 490 495

Gln Trp Thr Asp Arg Ile Met Glu Glu Phe Phe Arg Gln Gly Asp Arg

500 505 510

Glu Arg Glu Arg Gly Met Glu Ile Ser Pro Met Cys Asp Lys His Asn

515 520 525

Ala Ser Val Glu Lys Ser Gln Val Gly Phe Ile Asp Tyr Ile Val His

530 535 540

Pro Leu Trp Glu Thr Trp Ala Asp Leu Val His Pro Asp Ala Gln Asp

545 550 555 560

Ile Leu Asp Thr Leu Glu Asp Asn Arg Glu Trp Tyr Gln Ser Thr Ile

565 570 575

Pro Gln Ser Pro Ser Pro Ala Pro Asp Asp Pro Glu Glu Gly Arg Gln

580 585 590

Gly Gln Thr Glu Lys Phe Gln Phe Glu Leu Thr Leu Glu Glu Asp Gly

595 600 605

Glu Ser Asp Thr Glu Lys Asp Ser Gly Ser Gln Val Glu Glu Asp Thr

610 615 620

Ser Cys Ser Asp Ser Lys Thr Leu Cys Thr Gln Asp Ser Glu Ser Thr

625 630 635 640

Glu Ile Pro Leu Asp Glu Gln Val Glu Glu Glu Ala Val Gly Glu Glu

645 650 655

Glu Glu Ser Gln Pro Glu Ala Cys Val Ile Asp Asp Arg Ser Pro Asp

660 665 670

Thr

<210> 10

<211> 8240

<212> DNA

<213> Homo sapiens

<400> 10

cccctctcgg tagccctgag gctctggcgc cttcaagtga gaagctaagc accagcctct 60

gctgggctgc agaagcggcg gcggcggcag cagcagcagc agcatcagga aggcgctcgg 120

gccagcgcgg tgaacccgggg ctgggcagca ggtcgcggag ccgcgagcca ggatggaggc 180

agagggcagc agcgcgccgg cccgggcgggg cagcggagag ggcagcgaca gcgccggcgg 240

ggccacgctc aaagccccca agcatctctg gaggcacgag cagcaccacc agtacccgct 300

ccggcagccc cagttccgcc tcctgcatcc ccatcaccac ctgcccccgc cgccgccacc 360

ctcgccccag ccccagcccc agtgtccgct acagccgccg ccgccgcccc ccctgccgcc 420

gcccccgccg ccgcccgggg ctgcccgcgg ccgctacgcc tcgagcgggg ccaccggccg 480

cgtccggcat cgcggctact cggacaccga gcgctacctg tactgtcgcg ccatggaccg 540

cacctcctac gcggtggaga ccggccaccg gcccggcctg aagaaatcca ggatgtcctg 600

gccctcctcg ttccagggac tcaggcgttt tgatgtggac aatggcacat ctgcgggacg 660

gagtcccttg gatcccatga ccagcccagg atccgggcta attctccaag caaattttgt 720

ccacagtcaa cgacgggagt ccttcctgta tcgatccgac agcgattatg acctctctcc 780

aaagtctatg tcccggaact cctccattgc cagtgatata cacggagatg acttgattgt 840

gactccattt gctcaggtct tggccagtct gcgaactgta cgaaacaact ttgctgcatt 900

aactaatttg caagatcgag cacctagcaa aagatcaccc atgtgcaacc aaccatccat 960

caacaaagcc accataacag aggaggccta ccagaaactg gccagcgaga ccctggagga 1020

gctggactgg tgtctggacc agctagagac cctacagacc aggcactccg tcagtgagat 1080

ggcctccaac aagtttaaaa ggatgcttaa tcgggagctc acccatctct ctgaaatgag 1140

tcggtctgga aatcaagtgt cagagtttat atcaaacaca ttcttagata agcaacatga 1200

agtggaaatt ccttctccaa ctcagaagga aaaggagaaa aagaaaagac caatgtctca 1260

gatcagtgga gtcaagaaat tgatgcacag ctctagtctg actaattcaa gtatcccaag 1320

gtttggagtt aaaactgaac aagaagatgt ccttgccaag gaactagaag atgtgaacaa 1380

atggggtctt catgttttca gaatagcaga gttgtctggt aaccggccct tgactgttat 1440

catgcacacc atttttcagg aacgggattt attaaaaaca tttaaaattc cagtagatac 1500

tttaattaca tatcttatga ctctcgaaga ccattaccat gctgatgtgg cctatcacaa 1560

caatatccat gctgcagatg ttgtccagtc tactcatgtg ctattatcta cacctgcttt 1620

1680

tgtagatcat cctggtgtgt ccaatcaatt tctgatcaat acaaactctg aacttgcctt 1740

gatgtacaat gattcctcag tcttagagaa ccatcatttg gctgtggggct ttaaattgct 1800

tcaggaagaa aactgtgaca ttttccagaa tttgaccaaa aaacaaagac aatctttaag 1860

gaaaatggtc attgacatcg tacttgcaac agatatgtca aaacacatga atctactggc 1920

tgatttgaag actatggttg aaactaagaa agtgacaagc tctggagttc ttcttcttga 1980

taattattcc gataggattc aggttcttca gaatatggtg cactgtgcag atctgagcaa 2040

cccaacaaag cctctccagc tgtaccgcca gtggacggac cggataatgg aggagttctt 2100

ccgccaagga gaccgagaga gggaacgtgg catggagata agccccatgt gtgacaagca 2160

caatgcttcc gtggaaaaat cacaggtggg cttcatagac tatattgttc atcccctctg 2220

ggagacatgg gcagacctcg tccaccctga cgcccaggat attttggaca ctttggagga 2280

caatcgtgaa tggtaccaga gcacaatccc tcagagcccc tctcctgcac ctgatgaccc 2340

agaggagggc cggcagggtc aaactgagaa attccagttt gaactaactt tagaggaaga 2400

tggtgagtca gacacggaaa aggacagtgg cagtcaagtg gaagaagaca ctagctgcag 2460

tgactccaag actctttgta ctcaagactc agagtctact gaaattcccc ttgatgaaca 2520

ggttgaagag gaggcagtag gggaagaaga ggaaagccag cctgaagcct gtgtcataga 2580

tgatcgttct cctgacacgt aacagtgcaa aaactttcat gcctttttttt tttttaagta 2640

gaaaaattgt ttccaaagtg catgtcacat gccacaacca cggtcacacc tcactgtcat 2700

ctgccaggac gtttgttgaa caaaactgac cttgactact cagtccagcg ctcaggaata 2760

tcgtaaccag ttttttcacc tccatgtcat ccgagcaagg tggacatctt cacgaacagc 2820

gtttttaaca agatttcagc ttggtagagc tgacaaagca gtaaaatct actccaaatt 2880

attttcaaga gagtgtgact catcaggcag cccaaaagtt tattggactt ggggtttcta 2940

ttccttttta tttgtttgca atattttcag aagaaaggca ttgcacagag tgaacttaat 3000

ggacgaagca acaaatatgt caagaacagg acatagcacg aatctgttac cagtaggagg 3060

aggatgagcc acagaaattg cataattttc taatttcaag tcttcctgat acatgactga 3120

atagtgtggt tcagtgagct gcactgacct ctacattttg tatgatatgt aaaacagatt 3180

ttttgtagag cttactttta ttattaaatg tattgaggta ttatatttaa aaaaaactat 3240

gttcagaact tcatctgcca ctggttattt ttttctaagg agtaacttgc aagttttcag 3300

3360

ttcatagcaa ttaactgatt tgtagtgatt cattgtttgt tttatatacc aatgacttcc 3420

3480

atttactttg cattttgttt cactctttcc agataagcag agttgctctt caccagtgtt 3540

tttcttcatg tgcaaagtga ctatttgttc tataatactt ttatgtgtgt tatatcaaat 3600

gtgtcttaag cttcatgcaa actcagtcat cagttcgtgt tgtctgaagc aagtgggaga 3660

tatataaata cccagtagct aaaatggtca gtctttttta gatgttttcc tacttagtat 3720

ctcctaataa cgttttgctg tgtcactaga tgttcatttc acaagtgcat gtctttctaa 3780

taatccacac atttcatgct ctaataatcc acacatttca tgctcatttt tattgttttt 3840

acagccagtt atagtaagaa aaaggttttt ccccttgtgc tgctttataa tttagcgtgt 3900

3960 gtctgaacct tatccatgtt tgctagatga ggtcttgtca

accggtgaaa agaaacaggt agttaagtta gggttaacat tcatttcaac cacgaggttg 4020

tatatcatga ctagctttta ctcttggttt acagagaaaa gttaaacagc caactaggca 4080

gtttttaaga atattaacaa tatattaaca aacaccaata caactaatcc tatttggttt 4140

taatgatttc accatgggat taagaactat atcaggaaca tccctgagaa acggttttaa 4200

gtgtagcaac tactcttcct taatggacag ccacataacg tgtaggaagt cctttatcac 4260

ttatcctcga tccataagca tatcttgcag aggggaacta cttctttaaa cacatggagg 4320

gaaagaagat gatgccactg gcaccagagg gttagtactg tgatgcatcc taaaatattt 4380

attatattgg taaaaattct ggttaaataa aaaattagag atcactcttg gctgatttca 4440

gcaccaggaa ctgtattaca gttttagaga ttaattccta gtgtttacct gattatagca 4500

gttggcatca tggggcattt aattctgact ttatccccac gtcagcctta ataaagtctt 4560

ctttaccttc tctatgaaga ctttaaagcc caaataatca tttttcacat tgatattcaa 4620

gaattgagat agatagaagc caaagtgggt atctgacaag tggaaaatca aacgtttaag 4680

aagaattaca actctgaaaa gcatttatat gtggaacttc tcaaggagcc tcctggggac 4740

tggaaagtaa gtcatcagcc aggcaaatga ctcatgctga agagagtccc catttcagtc 4800

ccctgagatc tagctgatgc ttagatcctt tgaaataaaa attatgtctt tataactctg 4860

atcttttaca taaagcagaa gaggaatcaa ctagttaatt gcaaggtttc tactctgttt 4920

cctctgtaaa gatcagatgg taatctttca aataagaaaa aaataaagac gtatgtttga 4980

ccaagtagtt tcacaagaat atttgggaac ttgtttcttt taattttatt tgtccctgag 5040

tgaagtctag aaagaaaggt aaagagtcta gagtttattc ctctttccaa aacattctca 5100

ttcctctcct ccctacactt agtatttccc ccacagagtg ccctagaatct taataatgaa 5160

taaaataaaa agcagcaata tgtcattaac aaatccagac ctgaaagggt aaagggttta 5220

taactgcact aataaagaga ggctcttttt ttttcttcca gtttgttggt ttttaatggt 5280

accgtgttgt aaagataccc actaatggac aatcaaattg cagaaaaggc tcaatatcca 5340

agagacaggg actaatgcac tgtacaatct gcttatcctt gcccttctct cttgccaaag 5400

tgtgcttcag aaatatatac tgctttaaaa aagaataaaa gaatatcctt ttacaagtgg 5460

ctttacattt cctaaaatgc cataagaaaa tgcaatatct gggtactgta tggggaaaaa 5520

aatgtccaag tttgtgtaaa accagtgcat ttcagcttgc aagttactga acacaataat 5580

gctgttttaa ttttgtttta tatcagttaa aattcacaat aatgtagata gaacaaatta 5640

cagacaagga aagaaaaaac ttgaatgaaa tggattttac agaaagcttt atgataattt 5700

ttgaatgcat tatttatttt ttgtgccatg catttttttt ctcaccaaat gaccttacct 5760

gtaatacagt cttgtttgtc tgtttacaac catgtattta ttgcaatgta catactgtaa 5820

tgttaattgt aaattatctg ttcttattaa aacatcatcc catgatggga tggtgttgat 5880

atatttggaa actcttggtg agagaatgaa tggtgtgtat acatactctg tacatttttc 5940

6000

6060

ggctgtgcct taactttaac caatattttc ttctgttttg ctgcatttga aaggtaacag 6120

tggagctagg gctgggcatt ttacatccag gcttttaatt gattagaatt ctgccaaatag 6180

6240

gctcttaagt acttcttgcc agggagcagc actgcatgtg tgatggttgt ttgccatctg 6300

6360

tcggaaacac aactggggaa atatattctt tcccagtgat tataaacaat ctttttcttt 6420

tttttaagtc cttttggctt ctagagctca taggaaaatg gacttgattt gaaattggag 6480

ccagagttta ctcgtgttgg ttatctattc atcagcttcc tgacatgtta agagaataca 6540

ttaaagagaa aatactgttt tttaatccta aaatttttct tcactaaga taaaccaaat 6600

gtccttacat atatgtaaac ccatctattt aaacgcaaag gtgggttgat gtcagtttac 6660

atagcagaaa gcattcacta tcctctaaga tttgtttctg caaaactttc attgctttag 6720

6780

attatatgga aacatcctcc cagtacttgc ccagcccttg aatcatgtgg cttttcagtg 6840

6900

ttgacacaaa ctgtagattt tagcagccct ggcccaaagg aatttgatta cttttgtttt 6960

aaacagtaca aaggggacac tataattaca aaaacatcct taactgattt gagttgtttt 7020

tatttctttg gatatatttt cagagtggta aattgtgtgt gagaattaca aatgattatt 7080

cttttagtgg tttcttagcc tctcttacag cccacgggga tagtactgta catcaatacc 7140

ttcatatgaa atttttatat gcaatgaaaa taaaagcatg ggttgattct gcctatttat 7200

gactcaatct tttacaaata aaagattatt cattttaaat tatagttcaa tcagcatgtc 7260

tcttaggata ctgaacgtgg ttgaaatgaa aggatagtga catcataagt tagtactgat 7320

attcataacc aaataaagcc aacttgagta attttgctac attaaaaatt accaaaatta 7380

cttagatggc ctataagatt aagcatggtg ttttctaagc aagctttgaa aggggccttc 7440

catacttact taattgaata ttctgggata ttgaaaatta ttcagatact tgacaattat 7500

ttttggttac ctactccgca aactacaaag ttttaaggac tcaacaataa gttaatgaga 7560

cacagtgttt gctttcatgg agcttacagt ctggagggga caaaggctta aacaatactc 7620

atataattat atatgtgatc agtacaatga aggagctcag tggggtaaat aagcaggaac 7680

ctgaacttga tctgttccgg agggccacag aaggcttcct tgaggccttg agaaagtgat 7740

ttgcatctga gttctgaagg attgtaagag gtaactaggg aaaaagttga caggaagagg 7800

aaggggatcc agacaagaaa catttgcaaa gatcttgagg cataaatgag cttgagacat 7860

ctggagaaac tgaggaaaag tgagagagta ggcagggcct ggagccgcag agccattgct 7920

aaccatcctg tgtgagatat cccccattct gtagctttat tctcataacc ctgctcaatt 7980

ttctttataa cacttctcac agatttatat acgtgtttgt ttttgttatc tgtctctccc 8040

accagaccac agctccatga gagcaaggtc tttgcttacc aatatatcac tagcacttaa 8100

aactatgcct ggtacacagt aggttcttaa tatgtgttga atatagccat caaattgata 8160

ttggatataa ttcaatctga taagatattt tgagatatta aagagttttt aacttgatac 8220

cataaaaaaaaaaaaaaaaa 8240

<210> 11

<211> 809

<212> PROTEIN

<213> Homo sapiens

<400> 11

Met Glu Ala Glu Gly Ser Ser Ala Pro Ala Arg Ala Gly Ser Gly Glu

1 5 10 15

Gly Ser Asp Ser Ala Gly Gly Ala Thr Leu Lys Ala Pro Lys His Leu

20 25 30

Trp Arg His Glu Gln His His Gln Tyr Pro Leu Arg Gln Pro Gln Phe

35 40 45

Arg Leu Leu His Pro His His His Leu Pro Pro Pro Pro Pro Pro Ser

50 55 60

Pro Gln Pro Gln Pro Gln Cys Pro Leu Gln Pro Pro Pro Pro Pro Pro

65 70 75 80

Leu Pro Pro Pro Pro Pro Pro Pro Gly Ala Ala Arg Gly Arg Tyr Ala

85 90 95

Ser Ser Gly Ala Thr Gly Arg Val Arg His Arg Gly Tyr Ser Asp Thr

100 105 110

Glu Arg Tyr Leu Tyr Cys Arg Ala Met Asp Arg Thr Ser Tyr Ala Val

115 120 125

Glu Thr Gly His Arg Pro Gly Leu Lys Lys Ser Arg Met Ser Trp Pro

130 135 140

Ser Ser Phe Gln Gly Leu Arg Arg Phe Asp Val Asp Asn Gly Thr Ser

145 150 155 160

Ala Gly Arg Ser Pro Leu Asp Pro Met Thr Ser Pro Gly Ser Gly Leu

165 170 175

Ile Leu Gln Ala Asn Phe Val His Ser Gln Arg Arg Glu Ser Phe Leu

180 185 190

Tyr Arg Ser Asp Ser Asp Tyr Asp Leu Ser Pro Lys Ser Met Ser Arg

195 200 205

Asn Ser Ser Ile Ala Ser Asp Ile His Gly Asp Asp Leu Ile Val Thr

210 215 220

Pro Phe Ala Gln Val Leu Ala Ser Leu Arg Thr Val Arg Asn Asn Phe

225 230 235 240

Ala Ala Leu Thr Asn Leu Gln Asp Arg Ala Pro Ser Lys Arg Ser Pro

245 250 255

Met Cys Asn Gln Pro Ser Ile Asn Lys Ala Thr Ile Thr Glu Glu Ala

260 265 270

Tyr Gln Lys Leu Ala Ser Glu Thr Leu Glu Glu Leu Asp Trp Cys Leu

275 280 285

Asp Gln Leu Glu Thr Leu Gln Thr Arg His Ser Val Ser Glu Met Ala

290 295 300

Ser Asn Lys Phe Lys Arg Met Leu Asn Arg Glu Leu Thr His Leu Ser

305 310 315 320

Glu Met Ser Arg Ser Gly Asn Gln Val Ser Glu Phe Ile Ser Asn Thr

325 330 335

Phe Leu Asp Lys Gln His Glu Val Glu Ile Pro Ser Pro Thr Gln Lys

340 345 350

Glu Lys Glu Lys Lys Lys Arg Pro Met Ser Gln Ile Ser Gly Val Lys

355 360 365

Lys Leu Met His Ser Ser Ser Leu Thr Asn Ser Ser Ile Pro Arg Phe

370 375 380

Gly Val Lys Thr Glu Gln Glu Asp Val Leu Ala Lys Glu Leu Glu Asp

385 390 395 400

Val Asn Lys Trp Gly Leu His Val Phe Arg Ile Ala Glu Leu Ser Gly

405 410 415

Asn Arg Pro Leu Thr Val Ile Met His Thr Ile Phe Gln Glu Arg Asp

420 425 430

Leu Leu Lys Thr Phe Lys Ile Pro Val Asp Thr Leu Ile Thr Tyr Leu

435 440 445

Met Thr Leu Glu Asp His Tyr His Ala Asp Val Ala Tyr His Asn Asn

450 455 460

Ile His Ala Ala Asp Val Val Gln Ser Thr His Val Leu Leu Ser Thr

465 470 475 480

Pro Ala Leu Glu Ala Val Phe Thr Asp Leu Glu Ile Leu Ala Ala Ile

485 490 495

Phe Ala Ser Ala Ile His Asp Val Asp His Pro Gly Val Ser Asn Gln

500 505 510

Phe Leu Ile Asn Thr Asn Ser Glu Leu Ala Leu Met Tyr Asn Asp Ser

515 520 525

Ser Val Leu Glu Asn His His Leu Ala Val Gly Phe Lys Leu Leu Gln

530 535 540

Glu Glu Asn Cys Asp Ile Phe Gln Asn Leu Thr Lys Lys Gln Arg Gln

545 550 555 560

Ser Leu Arg Lys Met Val Ile Asp Ile Val Leu Ala Thr Asp Met Ser

565 570 575

Lys His Met Asn Leu Leu Ala Asp Leu Lys Thr Met Val Glu Thr Lys

580 585 590

Lys Val Thr Ser Ser Gly Val Leu Leu Leu Asp Asn Tyr Ser Asp Arg

595 600 605

Ile Gln Val Leu Gln Asn Met Val His Cys Ala Asp Leu Ser Asn Pro

610 615 620

Thr Lys Pro Leu Gln Leu Tyr Arg Gln Trp Thr Asp Arg Ile Met Glu

625 630 635 640

Glu Phe Phe Arg Gln Gly Asp Arg Glu Arg Glu Arg Gly Met Glu Ile

645 650 655

Ser Pro Met Cys Asp Lys His Asn Ala Ser Val Glu Lys Ser Gln Val

660 665 670

Gly Phe Ile Asp Tyr Ile Val His Pro Leu Trp Glu Thr Trp Ala Asp

675 680 685

Leu Val His Pro Asp Ala Gln Asp Ile Leu Asp Thr Leu Glu Asp Asn

690 695 700

Arg Glu Trp Tyr Gln Ser Thr Ile Pro Gln Ser Pro Ser Pro Ala Pro

705 710 715 720

Asp Asp Pro Glu Glu Gly Arg Gln Gly Gln Thr Glu Lys Phe Gln Phe

725 730 735

Glu Leu Thr Leu Glu Glu Asp Gly Glu Ser Asp Thr Glu Lys Asp Ser

740 745 750

Gly Ser Gln Val Glu Glu Asp Thr Ser Cys Ser Asp Ser Lys Thr Leu

755 760 765

Cys Thr Gln Asp Ser Glu Ser Thr Glu Ile Pro Leu Asp Glu Gln Val

770 775 780

Glu Glu Glu Ala Val Gly Glu Glu Glu Glu Ser Gln Pro Glu Ala Cys

785 790 795 800

Val Ile Asp Asp Arg Ser Pro Asp Thr

805

<210> 12

<211> 7979

<212> DNA

<213> Homo sapiens

<400> 12

cagcagcagg ctcagacctg cttccctggga catttccggg accgtgagcg agggaaccac 60

gttgccctgg attcttgcca gctgtacaaa gttgaccagg aaaatggctc agcagacaag 120

cccggacact ttaacagtac ctgaagtgga taatccgcat tgtccaaacc cgtggctgaa 180

cgaagacctt gtgaaatcct tgcgagaaaa cctgttgcag catgagaagt ccaagacagc 240

gaggaaatcg gtttctccca agctctctcc agtgatctct ccgagaaatt cccccaggct 300

tctgcgcaga atgcttctca gcagcaacat ccccaaacag cggcgtttca cggtggcaca 360

tacatgtttt gatgtggaca atggcacatc tgcgggacgg agtcccttgg atcccatgac 420

cagccgga tccgggctaa ttctccaagc aaattttgtc cacagtcaac gacgggagtc 480

cttcctgtat cgatccgaca gcgattatga cctctctcca aagtctatgt ccgggaactc 540

ctccattgcc agtgatatac acggagatga cttgattgtg actccatttg ctcaggtctt 600

ggccagtctg cgaactgtac gaaacaactt tgctgcatta actaatttgc aagatcgagc 660

acctagcaaa agatcaccca tgtgcaacca accatccatc aacaaagcca ccataacaga 720

ggaggcctac cagaaactgg ccagcgagac cctggaggag ctggactggt gtctggacca 780

gctagagacc ctacagacca ggcactccgt cagtgagatg gcctccaaca agtttaaaag 840

gatgcttaat cgggagctca cccatctctc tgaaatgagt cggtctggaa atcaagtgtc 900

agagtttata tcaaacacat tcttagataa gcaacatgaa gtggaaattc cttctccaac 960

tcagaaggaa aaggagaaaa agaaaagacc aatgtctcag atcagtggag tcaagaaatt 1020

gatgcacagc tctagtctga ctaattcaag tatcccaagg tttggagtta aaactgaaca 1080

agaagatgtc cttgccaagg aactagaaga tgtgaacaaa tggggtcttc atgttttcag 1140

aatagcagag ttgtctggta accggccctt gactgttatc atgcacacca tttttcagga 1200

acgggattta ttaaaaacat ttaaaattcc agtagatact ttaattacat atcttatgac 1260

tctcgaagac cattaccatg ctgatgtggc ctatcacaac aatatccatg ctgcagatgt 1320

tgtccagtct actcatgtgc tattatctac acctgctttg gaggctgtgt ttacagattt 1380

ggagattctt gcagcaattt ttgccagtgc aatacatgat gtagatcatc ctggtgtgtc 1440

caatcaattt ctgatcaata caaactctga acttgccttg atgtacaatg attcctcagt 1500

cttagagaac catcatttgg ctgtgggctt taaattgctt caggaagaaa actgtgacat 1560

tttccagaat ttgaccaaaa aacaaagaca atctttaagg aaaatggtca ttgacatcgt 1620

acttgcaaca gatatgtcaa aacacatgaa tctactggct gatttgaaga ctatggttga 1680

aactaagaaa gtgacaagct ctggagttct tcttcttgat aattattccg ataggattca 1740

ggttcttcag aatatggtgc actgtgcaga tctgagcaac ccaacaaagc ctctccagct 1800

gtaccgccag tggacggacc ggataatgga ggagttcttc cgccaaggag accgagagag 1860

ggaacgtggc atggagataa gccccatgtg tgacaagcac aatgcttccg tggaaaaatc 1920

acaggtgggc ttcatagact atattgttca tcccctctgg gagacatggg cagacctcgt 1980

ccaccctgac gcccaggata ttttggacac tttggaggac aatcgtgaat ggtaccagag 2040

cacaatccct cagagcccct ctcctgcacc tgatgaccca gaggagggcc ggcagggtca 2100

2160

ggacagtggc agtcaagtgg aagaagacac tagctgcagt gactccaaga ctctttgtac 2220

tcaagactca gagtctactg aaattcccct tgatgaacag gttgaagagg aggcagtagg 2280

ggaagaagag gaaagccagc ctgaagcctg tgtcatagat gatcgttctc ctgacacgta 2340

acagtgcaaa aactttcatg cctttttttt ttttaagtag aaaaattgtt tccaaagtgc 2400

atgtcacatg ccacaaccac ggtcacacct cactgtcatc tgccaggacg tttgttgaac 2460

aaaactgacc ttgactactc agtccagcgc tcaggaatat cgtaaccagt tttttcacct 2520

ccatgtcatc cgagcaaggt ggacatcttc acgaacagcg tttttaacaa gatttcagct 2580

tggtagagct gacaaagcag ataaaatcta ctccaaatta ttttcaagag agtgtgactc 2640

atcaggcagc ccaaaagttt attggacttg gggtttctat tcctttttat ttgtttgcaa 2700

tattttcaga agaaaggcat tgcacagagt gaacttaatg gacgaagcaa caaatatgtc 2760

aagaacagga catagcacga atctgttacc agtaggagga ggatgagcca cagaaattgc 2820

ataattttct aatttcaagt cttcctgata catgactgaa tagtgtggtt cagtgagctg 2880

2940

tattaaatgt attgaggtat tatatttaaa aaaaactatg ttcagaactt catctgccac 3000

3060

ataaaaatct aatttatgaa ttttacttgc accttatagt tcatagcaat taactgattt 3120

3180

actttatgtt gcaggaaacc ctttttgtaa gtctttatta tttactttgc attttgtttc 3240

actctttcca gataagcaga gttgctcttc accagtgttt ttcttcatgt gcaaagtgac 3300

3360

ctcagtcatc agttcgtgtt gtctgaagca agtgggagat atataaatac ccagtagcta 3420

aaatggtcag tcttttttag atgttttcct acttagtatc tcctaataac gttttgctgt 3480

gtcactagat gttcatttca caagtgcatg tctttctaat aatccacaca tttcatgctc 3540

taataatcca cacatttcat gctcattttt attgttttta cagccagtta tagtaagaaa 3600

aaggtttttc cccttgtgct gctttataat ttagcgtgtg tctgaacctt atccatgttt 3660

gctagatgag gtcttgtcaa atatatcact accattgtca ccggtgaaaa gaaacaggta 3720

gttaagttag ggttaacatt catttcaacc acgaggttgt atatcatgac tagcttttac 3780

3840

atattaacaa acaccaatac aactaatcct atttggtttt aatgatttca ccatgggatt 3900

aagaactata tcaggaacat ccctgagaaa cggttttaag tgtagcaact actcttcctt 3960

4020

atcttgcaga ggggaactac ttctttaaac acatggaggg aaagaagatg atgccactgg 4080

caccagaggg ttagtactgt gatgcatcct aaaatattta ttatattggt aaaaattctg 4140

gttaaataaa aaattagaga tcactcttgg ctgatttcag caccaggaac tgtattacag 4200

ttttagagat taattcctag tgtttacctg attatagcag ttggcatcat ggggcattta 4260

attctgactt tatccccacg tcagccttaa taaagtcttc tttaccttct ctatgaagac 4320

aattgagata gatagaagcc 4380

aaagtgggta tctgacaagt ggaaaatcaa acgtttaaga agaattacaa ctctgaaaag 4440

catttatatg tggaacttct caaggagcct cctggggact ggaaagtaag tcatcagcca 4500

ggcaaatgac tcatgctgaa gagagtcccc atttcagtcc cctgagatct agctgatgct 4560

tagatccttt gaaataaaaa ttatgtcttt ataactctga tcttttacat aaagcagaag 4620

aggaatcaac tagttaattg caaggtttct actctgtttc ctctgtaaag atcagatggt 4680

4740

tttgggaact tgtttctttt aattttattt gtccctgagt gaagtctaga aagaaaggta 4800

aagagtctag agtttattcc tctttccaaa acattctcat tcctctcctc cctacactta 4860

gtatttcccc cacagagtgc ctagaatctt aataatgaat aaaataaaaa gcagcaatat 4920

gtcattaaca aatccagacc tgaaagggta aagggtttat aactgcacta ataaagagag 4980

gctctttttt ttttcttccag tttgttggtt tttaatggta ccgtgttgta aagataccca 5040

ctaatggaca atcaaattgc agaaaaggct caatatccaa gagacaggga ctaatgcact 5100

gtacaatctg cttatccttg cccttctctc ttgccaaagt gtgcttcaga aatatatact 5160

gctttaaaaa agaataaaag aatatccttt tacaagtggc tttacatttc ctaaaatgcc 5220

ataagaaaat gcaatatctg ggtactgtat ggggaaaaaa atgtccaagt ttgtgtaaaa 5280

ccagtgcatt tcagcttgca agttactgaa cacaataatg ctgttttaat tttgttttat 5340

atcagttaaa attcacaata atgtagatag aacaaattac agacaaggaa agaaaaaact 5400

5460

tgtgccatgc attttttttc tcaccaaatg accttacctg taatacagtc ttgtttgtct 5520

5580

tcttattaaa acatcatccc atgatgggat ggtgttgata tatttggaaa ctcttggtga 5640

gagaatgaat ggtgtgtata catactctgt acatttttct tttctcctgt aatatagtct 5700

5760

taaatttaaa aaaacatagc tgcaggtctt tggtcccagg gctgtgcctt aactttaacc 5820

aatattttct tctgttttgc tgcatttgaa aggtaacagt ggagctaggg ctgggcattt 5880

tacatccagg cttttaattg attagaattc tgccaatagg tggattttac aaaaccacag 5940

acaacctctg aaagattctg agaccctttt gagacagaag ctcttaagta cttcttgcca 6000

gggagcagca ctgcatgtgt gatggttgtt tgccatctgt tgatcaggaa ctacttcagc 6060

6120

6180

tagagctcat aggaaaatgg acttgatttg aaattggagc cagagtttac tcgtgttggt 6240

tatctattca tcagcttcct gacatgttaa gagaatacat taaagagaaa atactgtttt 6300

ttaatcctaa aatttttctt ccactaagat aaaccaaatg tccttacata tatgtaaacc 6360

catctattta aacgcaaagg tgggttgatg tcagtttaca tagcagaaag cattcactat 6420

6480

acaatggcca gcccctaaag caggaaacat ttataatgga ttatatggaa acatcctccc 6540

agtacttgcc cagcccttga atcatgtggc ttttcagtga aaggaaagat tctttttcta 6600

6660

agcagccctg gcccaaagga atttgattac ttttgtttta aacagtacaa aggggacact 6720

ataattacaa aaacatcctt aactgatttg agttgttttt atttctttgg atatattttc 6780

agagtggtaa attgtgtgtg agaattacaa atgattattc ttttagtggt ttcttagcct 6840

ctcttacagc ccacggggat agtactgtac atcaatacct tcatatgaaa tttttatatg 6900

caatgaaaat aaaagcatgg gttgattctg cctatttatg actcaatctt ttacaaataa 6960

aagattattc attttaaatt atagttcaat cagcatgtct cttaggatac tgaacgtggt 7020

tgaaatgaaa ggatagtgac atcataagtt agtactgata ttcataacca aataaagcca 7080

acttgagtaa ttttgctaca ttaaaaatta ccaaaattac ttagatggcc tataagatta 7140

agcatggtgt tttctaagca agctttgaaa ggggccttcc atacttactt aattgaatat 7200

tctgggatat tgaaaattat tcagatactt gacaattatt tttggttacc tactccgcaa 7260

actacaaagt tttaaggact caacaataag ttaatgagac acagtgtttg ctttcatgga 7320

gcttacagtc tggaggggac aaaggcttaa acaatactca tataattata tatgtgatca 7380

gtacaatgaa ggagctcagt ggggtaaata agcaggaacc tgaacttgat ctgttccgga 7440

gggccacaga aggcttcctt gaggccttga gaaagtgatt tgcatctgag ttctgaagga 7500

7560

atttgcaaag atcttgaggc ataaatgagc ttgagacatc tggagaaact gaggaaaagt 7620

gagagagtag gcagggcctg gagccgcaga gccattgcta accatcctgt gtgagatatc 7680

ccccattctg tagctttatt ctcataaccc tgctcaattt tctttataac acttctcaca 7740

gatttatata cgtgtttgtt tttgttatct gtctctccca ccagaccaca gctccatgag 7800

agcaaggtct ttgcttacca atatatcact agcacttaaa actatgcctg gtacacagta 7860

ggttcttaat atgtgttgaa tatagccatc aaattgatat tggatataat tcaatctgat 7920

aagatatttt gagatattaa agagttttta acttgatacc ataaaaaaaa aaaaaaaaa 7979

<210> 13

<211> 745

<212> PROTEIN

<213> Homo sapiens

<400> 13

Met Ala Gln Gln Thr Ser Pro Asp Thr Leu Thr Val Pro Glu Val Asp

1 5 10 15

Asn Pro His Cys Pro Asn Pro Trp Leu Asn Glu Asp Leu Val Lys Ser

20 25 30

Leu Arg Glu Asn Leu Leu Gln His Glu Lys Ser Lys Thr Ala Arg Lys

35 40 45

Ser Val Ser Pro Lys Leu Ser Pro Val Ile Ser Pro Arg Asn Ser Pro

50 55 60

Arg Leu Leu Arg Arg Met Leu Leu Ser Ser Asn Ile Pro Lys Gln Arg

65 70 75 80

Arg Phe Thr Val Ala His Thr Cys Phe Asp Val Asp Asn Gly Thr Ser

85 90 95

Ala Gly Arg Ser Pro Leu Asp Pro Met Thr Ser Pro Gly Ser Gly Leu

100 105 110

Ile Leu Gln Ala Asn Phe Val His Ser Gln Arg Arg Glu Ser Phe Leu

115 120 125

Tyr Arg Ser Asp Ser Asp Tyr Asp Leu Ser Pro Lys Ser Met Ser Arg

130 135 140

Asn Ser Ser Ile Ala Ser Asp Ile His Gly Asp Asp Leu Ile Val Thr

145 150 155 160

Pro Phe Ala Gln Val Leu Ala Ser Leu Arg Thr Val Arg Asn Asn Phe

165 170 175

Ala Ala Leu Thr Asn Leu Gln Asp Arg Ala Pro Ser Lys Arg Ser Pro

180 185 190

Met Cys Asn Gln Pro Ser Ile Asn Lys Ala Thr Ile Thr Glu Glu Ala

195 200 205

Tyr Gln Lys Leu Ala Ser Glu Thr Leu Glu Glu Leu Asp Trp Cys Leu

210 215 220

Asp Gln Leu Glu Thr Leu Gln Thr Arg His Ser Val Ser Glu Met Ala

225 230 235 240

Ser Asn Lys Phe Lys Arg Met Leu Asn Arg Glu Leu Thr His Leu Ser

245 250 255

Glu Met Ser Arg Ser Gly Asn Gln Val Ser Glu Phe Ile Ser Asn Thr

260 265 270

Phe Leu Asp Lys Gln His Glu Val Glu Ile Pro Ser Pro Thr Gln Lys

275 280 285

Glu Lys Glu Lys Lys Lys Arg Pro Met Ser Gln Ile Ser Gly Val Lys

290 295 300

Lys Leu Met His Ser Ser Ser Leu Thr Asn Ser Ser Ile Pro Arg Phe

305 310 315 320

Gly Val Lys Thr Glu Gln Glu Asp Val Leu Ala Lys Glu Leu Glu Asp

325 330 335

Val Asn Lys Trp Gly Leu His Val Phe Arg Ile Ala Glu Leu Ser Gly

340 345 350

Asn Arg Pro Leu Thr Val Ile Met His Thr Ile Phe Gln Glu Arg Asp

355 360 365

Leu Leu Lys Thr Phe Lys Ile Pro Val Asp Thr Leu Ile Thr Tyr Leu

370 375 380

Met Thr Leu Glu Asp His Tyr His Ala Asp Val Ala Tyr His Asn Asn

385 390 395 400

Ile His Ala Ala Asp Val Val Gln Ser Thr His Val Leu Leu Ser Thr

405 410 415

Pro Ala Leu Glu Ala Val Phe Thr Asp Leu Glu Ile Leu Ala Ala Ile

420 425 430

Phe Ala Ser Ala Ile His Asp Val Asp His Pro Gly Val Ser Asn Gln

435 440 445

Phe Leu Ile Asn Thr Asn Ser Glu Leu Ala Leu Met Tyr Asn Asp Ser

450 455 460

Ser Val Leu Glu Asn His His Leu Ala Val Gly Phe Lys Leu Leu Gln

465 470 475 480

Glu Glu Asn Cys Asp Ile Phe Gln Asn Leu Thr Lys Lys Gln Arg Gln

485 490 495

Ser Leu Arg Lys Met Val Ile Asp Ile Val Leu Ala Thr Asp Met Ser

500 505 510

Lys His Met Asn Leu Leu Ala Asp Leu Lys Thr Met Val Glu Thr Lys

515 520 525

Lys Val Thr Ser Ser Gly Val Leu Leu Leu Asp Asn Tyr Ser Asp Arg

530 535 540

Ile Gln Val Leu Gln Asn Met Val His Cys Ala Asp Leu Ser Asn Pro

545 550 555 560

Thr Lys Pro Leu Gln Leu Tyr Arg Gln Trp Thr Asp Arg Ile Met Glu

565 570 575

Glu Phe Phe Arg Gln Gly Asp Arg Glu Arg Glu Arg Gly Met Glu Ile

580 585 590

Ser Pro Met Cys Asp Lys His Asn Ala Ser Val Glu Lys Ser Gln Val

595 600 605

Gly Phe Ile Asp Tyr Ile Val His Pro Leu Trp Glu Thr Trp Ala Asp

610 615 620

Leu Val His Pro Asp Ala Gln Asp Ile Leu Asp Thr Leu Glu Asp Asn

625 630 635 640

Arg Glu Trp Tyr Gln Ser Thr Ile Pro Gln Ser Pro Ser Pro Ala Pro

645 650 655

Asp Asp Pro Glu Glu Gly Arg Gln Gly Gln Thr Glu Lys Phe Gln Phe

660 665 670

Glu Leu Thr Leu Glu Glu Asp Gly Glu Ser Asp Thr Glu Lys Asp Ser

675 680 685

Gly Ser Gln Val Glu Glu Asp Thr Ser Cys Ser Asp Ser Lys Thr Leu

690 695 700

Cys Thr Gln Asp Ser Glu Ser Thr Glu Ile Pro Leu Asp Glu Gln Val

705 710 715 720

Glu Glu Glu Ala Val Gly Glu Glu Glu Glu Ser Gln Pro Glu Ala Cys

725 730 735

Val Ile Asp Asp Arg Ser Pro Asp Thr

740 745

<210> 14

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> PDE4D5_forward primer

<400> 14

gcttctcagc agcaacatc 19

<210> 15

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> PDE4D5_reverse primer

<400> 15

tgccattgtc cacatcaaaa 20

<210> 16

<211> 24

<212> DNA

<213> Artificial sequence

<220>

<223> PDE4D5 probe

<400> 16

acagcggcgt ttcacggtgg caca 24

<210> 17

<211> 7591

<212> DNA

<213> Homo sapiens

<400> 17

agttccttat ttggtagctt ttgacaggac tagcctttct tgcaactaag catcttgaca 60

tacattattc attaagccct ggagctcggg agagaaagat gcagaccctt agatctttag 120

atattccttt atcacgtgga ttttctttat tcagaatagt tgctgaattt tgtgccattc 180

tggagtctta caaatggcat gtattcgatg ggaagacggc tggatgggat ttaatgcgag 240

gctttcttat gtatacttaa ttaccaaaaa tctttaaaaa ctcatactct gcgtggcttg 300

360

atacatatat atatatatac atataatcaa tcaaaaatgc ctgaagcaaa ctatttactg 420

tcagtgtctt ggggctacat aaagtttaaa aggatgctta atcgggagct cacccatctc 480

tctgaaatga gtcggtctgg aaatcaagtg tcagagttta tatcaaacac attcttagat 540

aagcaacatg aagtggaaat tccttctcca actcagaagg aaaaggagaa aaagaaaaga 600

ccaatgtctc agatcagtgg agtcaagaaa ttgatgcaca gctctagtct gactaattca 660

agtatcccaa ggtttggagt taaaactgaa caagaagatg tccttgccaa ggaactagaa 720

gatgtgaaca aatggggtct tcatgttttc agaatagcag agttgtctgg taaccggccc 780

ttgactgtta tcatgcacac catttttcag gaacgggatt tattaaaaac atttaaaatt 840

ccagtagata ctttaattac atatcttatg actctcgaag accattacca tgctgatgtg 900

gcctatcaca acaatatcca tgctgcagat gttgtccagt ctactcatgt gctattatct 960

1020

gcaatacatg atgtagatca tcctggtgtg tccaatcaat ttctgatcaa tacaaactct 1080

gaacttgcct tgatgtacaa tgattcctca gtcttagaga accatcattt ggctgtgggc 1140

tttaaattgc ttcaggaaga 1200

caatctttaa ggaaaatggt cattgacatc gtacttgcaa cagatatgtc aaaacacatg 1260

aatctactgg ctgatttgaa gactatggtt gaaactaaga aagtgacaag ctctggagtt 1320

cttcttcttg ataattattc cgataggatt caggttcttc agaatatggt gcactgtgca 1380

gatctgagca acccaacaaa gcctctccag ctgtaccgcc agtggacgga ccggataatg 1440

gaggagttct tccgccaagg agaccgagag agggaacgtg gcatggagat aagccccatg 1500

tgtgacaagc acaatgcttc cgtggaaaaa tcacaggtgg gcttcataga ctatattgtt 1560

catcccctct gggagacatg ggcagacctc gtccaccctg acgccccagga tattttggac 1620

actttggagg acaatcgtga atggtaccag agcacaatcc ctcagagccc ctctcctgca 1680

cctgatgacc cagaggaggg ccggcagggt caaactgaga aattccagtt tgaactaact 1740

1800

actagctgca gtgactccaa gactctttgt actcaagact cagagtctac tgaaattccc 1860

cttgatgaac aggttgaaga ggaggcagta ggggaagaag aggaaagcca gcctgaagcc 1920

tgtgtcatag atgatcgttc tcctgacacg taacagtgca aaaactttca tgcctttttt 1980

ttttttaagt agaaaaattg tttccaaagt gcatgtcaca tgccacaacc acggtcacac 2040

ctcactgtca tctgccagga cgtttgttga acaaaactga ccttgactac tcagtccagc 2100

gctcaggaat atcgtaacca gttttttcac ctccatgtca tccgagcaag gtggacatct 2160

tcacgaacag cgtttttaac aagatttcag cttggtagag ctgacaaagc agataaaatc 2220

2280

2340

gtgaacttaa tggacgaagc aacaaatatg tcaagaacag gacatagcac gaatctgtta 2400

ccagtaggag gaggatgagc cacagaaatt gcataatttt ctaatttcaa gtcttcctga 2460

tacatgactg aatagtgtgg ttcagtgagc tgcactgacc tctacatttt gtatgatatg 2520

2580

aaaaaaacta tgttcagaac ttcatctgcc actggttatt tttttctaag gagtaacttg 2640

caagttttca gtacaaatct gtgctacact ggataaaaat ctaatttatg aattttactt 2700

2760

caatgacttc catattttaa aagagaaaaa caactttatg ttgcaggaaa ccctttttgt 2820

aagtctttat tatttacttt gcattttgtt tcactctttc cagataagca gagttgctct 2880

tcaccagtgt ttttcttcat gtgcaaagtg actatttgtt ctataatact tttatgtgtg 2940

ttatatcaaa tgtgtcttaa gcttcatgca aactcagtca tcagttcgtg ttgtctgaag 3000

3060

ctacttagta tctcctaata acgttttgct gtgtcactag atgttcattt cacaagtgca 3120

tgtctttcta ataatccaca catttcatgc tctaataatc cacacatttc atgctcattt 3180

ttattgtttt tacagccagt tatagtaaga aaaaggtttt tccccttgtg ctgcttttata 3240

atttagcgtg tgtctgaacc ttatccatgt ttgctagatg aggtcttgtc aaatatatca 3300

ctaccattgt caccggtgaa aagaaacagg tagttaagtt agggttaaca ttcatttcaa 3360

ccacgaggtt gtatatcatg actagctttt actcttggtt tacagagaaa agttaaacag 3420

ccaactaggc agtttttaag aatattaaca atatattaac aaacaccaat acaactaatc 3480

3540

aacggtttta agtgtagcaa ctactcttcc ttaatggaca gccacataac gtgtaggaag 3600

3660

acacatggag ggaaagaaga tgatgccact ggcaccagag ggttagtact gtgatgcatc 3720

ctaaaatatt gtaaaaattc tggttaaata aaaaattaga gatcactctt 3780

ggctgatttc agcaccagga actgtattac agttttagag attaattcct agtgtttacc 3840

tgattatagc agttggcatc atggggcatt taattctgac tttatcccca cgtcagcctt 3900

aataaagtct tctttacctt ctctatgaag actttaaagc ccaaataatc atttttcaca 3960

ttgatattca agaattgaga tagatagaag ccaaagtggg tatctgacaa gtggaaaatc 4020

aaacgtttaa gaagaattac aactctgaaa agcatttata tgtggaactt ctcaaggagc 4080

ctcctgggga ctggaaagta agtcatcagc caggcaaatg actcatgctg aagagagtcc 4140

ccatttcagt cccctgagat ctagctgatg cttagatcct ttgaaataaa aattatgtct 4200

ttataactct gatcttttac ataaagcaga agaggaatca actagttaat tgcaaggttt 4260

ctactctgtt tcctctgtaa agatcagatg gtaatctttc aaataagaaa aaaataaaga 4320

4380

ttgtccctga gtgaagtcta gaaagaaagg taaagagtct agagtttatt cctctttcca 4440

aaacattctc attcctctcc tccctacact tagtatttcc cccacagagt gcctagaatc 4500

ttaataatga ataaaataaa aagcagcaat atgtcattaa caaatccaga cctgaaaggg 4560

taaagggttt ataactgcac taataaagag aggctctttt ttttttcttcc agtttgttgg 4620

tttttaatgg taccgtgttg taaagatacc cactaatgga caatcaaatt gcagaaaagg 4680

ctcaatatcc aagagacagg gactaatgca ctgtacaatc tgcttatcct tgcccttctc 4740

tcttgccaaa gtgtgcttca gaaatatata ctgctttaaa aaagaataaa agaatatcct 4800

tttacaagtg gctttacatt tcctaaaatg ccataagaaa atgcaatatc tgggtactgt 4860

atggggaaaa aaatgtccaa gtttgtgtaa aaccagtgca tttcagcttg caagttactg 4920

aacacaataa tgctgtttta attttgtttt atatcagtta aaattcacaa taatgtagat 4980

agaacaaatt acagacaagg aaagaaaaaa cttgaatgaa atggatttta cagaaagctt 5040

tatgataatt tttgaatgca ttatttattt tttgtgccat gcattttttt tctcaccaaa 5100

tgaccttacc tgtaatacag tcttgtttgt ctgtttacaa ccatgtattt attgcaatgt 5160

acatactgta atgttaattg taaattatct gttcttatta aaacatcatc ccatgatggg 5220

atggtgttga tatatttgga aactcttggt gagagaatga atggtgtgta tacatactct 5280

gtacattttt cttttctcct gtaatatagt cttgtcacct tagagcttgt ttatggaaga 5340

ttcaagaaaa ctataaaata cttaaagata tataaattta aaaaaacata gctgcaggtc 5400

tttggtccca gggctgtgcc ttaactttaa ccaatatttt cttctgtttt gctgcatttg 5460

aaaggtaaca gtggagctag ggctgggcat tttacatcca ggcttttaat tgattagaat 5520

tctgccaata ggtggatttt acaaaaccac agacaacctc tgaaagattc tgagaccctt 5580

ttgagacaga agctcttaag tacttcttgc cagggagcag cactgcatgt gtgatggttg 5640

tttgccatct gttgatcagg aactacttca gctacttgca tttgattatt tccttttttt 5700

ttttttttaa ctcggaaaca caactgggga aatatattct ttcccagtga ttataaacaa 5760

tctttttctt tttttttaagt ccttttggct tctagagctc ataggaaaat ggacttgatt 5820

tgaaattgga gccagagttt actcgtgttg gttatctatt catcagcttc ctgacatgtt 5880

aagagaatac attaaagaga aaatactgtt ttttaatcct aaaatttttc ttcactaag 5940

ataaaccaaa tgtccttaca tatatgtaaa cccatctatt taaacgcaaa ggtggggttga 6000

tgtcagttta catagcagaa agcattcact atcctctaag atttgtttct gcaaaacttt 6060

cattgcttta gaattttaaa atttcacctt gtacaatggc cagcccctaa agcaggaaac 6120

atttataatg gattatatgg aaacatcctc ccagtacttg cccagccctt gaatcatgtg 6180

6240

tattttattt tttgacacaa actgtagatt ttagcagccc tggcccaaag gaatttgatt 6300

acttttgttt taaacagtac aaaggggaca ctataattac aaaaacatcc ttaactgatt 6360

tgagttgttt ttatttcttt ggatatattt tcagagtggt aaattgtgtg tgagaattac 6420

aaatgattat tcttttagtg gtttcttagc ctctcttaca gcccacgggg atagtactgt 6480

acatcaatac cttcatatga aatttttata tgcaatgaaa ataaaagcat gggttgattc 6540

tgcctattta tgactcaatc ttttacaaat aaaagattat tcattttaaa ttatagttca 6600

atcagcatgt ctcttaggat actgaacgtg gttgaaatga aaggatagtg acatcataag 6660

6720

taccaaaatt acttagatgg cctataagat taagcatggt gttttctaag caagctttga 6780

aaggggcctt ccatacttac ttaattgaat attctgggat attgaaaatt attcagatac 6840

6900

agttaatgag acacagtgtt tgctttcatg gagcttacag tctggagggg acaaaggctt 6960

aaacaatact catataatta tatatgtgat cagtacaatg aaggagctca gtggggtaaa 7020

taagcaggaa cctgaacttg atctgttccg gagggccaca gaaggcttcc ttgaggcctt 7080

7140

acaggaagag gaaggggatc cagacaagaa acatttgcaa agatcttgag gcataaatga 7200

7260

gagccattgc taaccatcct gtgtgagata tcccccattc tgtagcttta ttctcataac 7320

cctgctcaat tttctttata acacttctca cagatttata tacgtgtttg tttttgttat 7380

ctgtctctcc caccagacca cagctccatg agagcaaggt ctttgcttac caatatatca 7440

ctagcactta aaactatgcc tggtacacag taggttctta atatgtgttg aatatagcca 7500

7560

taacttgata ccataaaaaa aaaaaaaaaaa 7591

<210> 18

<211> 518

<212> PROTEIN

<213> Homo sapiens

<400> 18

Met Pro Glu Ala Asn Tyr Leu Leu Ser Val Ser Trp Gly Tyr Ile Lys

1 5 10 15

Phe Lys Arg Met Leu Asn Arg Glu Leu Thr His Leu Ser Glu Met Ser

20 25 30

Arg Ser Gly Asn Gln Val Ser Glu Phe Ile Ser Asn Thr Phe Leu Asp

35 40 45

Lys Gln His Glu Val Glu Ile Pro Ser Pro Thr Gln Lys Glu Lys Glu

50 55 60

Lys Lys Lys Arg Pro Met Ser Gln Ile Ser Gly Val Lys Lys Leu Met

65 70 75 80

His Ser Ser Ser Leu Thr Asn Ser Ser Ile Pro Arg Phe Gly Val Lys

85 90 95

Thr Glu Gln Glu Asp Val Leu Ala Lys Glu Leu Glu Asp Val Asn Lys

100 105 110

Trp Gly Leu His Val Phe Arg Ile Ala Glu Leu Ser Gly Asn Arg Pro

115 120 125

Leu Thr Val Ile Met His Thr Ile Phe Gln Glu Arg Asp Leu Leu Lys

130 135 140

Thr Phe Lys Ile Pro Val Asp Thr Leu Ile Thr Tyr Leu Met Thr Leu

145 150 155 160

Glu Asp His Tyr His Ala Asp Val Ala Tyr His Asn Asn Ile His Ala

165 170 175

Ala Asp Val Val Gln Ser Thr His Val Leu Leu Ser Thr Pro Ala Leu

180 185 190

Glu Ala Val Phe Thr Asp Leu Glu Ile Leu Ala Ala Ile Phe Ala Ser

195 200 205

Ala Ile His Asp Val Asp His Pro Gly Val Ser Asn Gln Phe Leu Ile

210 215 220

Asn Thr Asn Ser Glu Leu Ala Leu Met Tyr Asn Asp Ser Ser Val Leu

225 230 235 240

Glu Asn His His Leu Ala Val Gly Phe Lys Leu Leu Gln Glu Glu Asn

245 250 255

Cys Asp Ile Phe Gln Asn Leu Thr Lys Lys Gln Arg Gln Ser Leu Arg

260 265 270

Lys Met Val Ile Asp Ile Val Leu Ala Thr Asp Met Ser Lys His Met

275 280 285

Asn Leu Leu Ala Asp Leu Lys Thr Met Val Glu Thr Lys Lys Val Thr

290 295 300

Ser Ser Gly Val Leu Leu Leu Asp Asn Tyr Ser Asp Arg Ile Gln Val

305 310 315 320

Leu Gln Asn Met Val His Cys Ala Asp Leu Ser Asn Pro Thr Lys Pro

325 330 335

Leu Gln Leu Tyr Arg Gln Trp Thr Asp Arg Ile Met Glu Glu Phe Phe

340 345 350

Arg Gln Gly Asp Arg Glu Arg Glu Arg Gly Met Glu Ile Ser Pro Met

355 360 365

Cys Asp Lys His Asn Ala Ser Val Glu Lys Ser Gln Val Gly Phe Ile

370 375 380

Asp Tyr Ile Val His Pro Leu Trp Glu Thr Trp Ala Asp Leu Val His

385 390 395 400

Pro Asp Ala Gln Asp Ile Leu Asp Thr Leu Glu Asp Asn Arg Glu Trp

405 410 415

Tyr Gln Ser Thr Ile Pro Gln Ser Pro Ser Pro Ala Pro Asp Asp Pro

420 425 430

Glu Glu Gly Arg Gln Gly Gln Thr Glu Lys Phe Gln Phe Glu Leu Thr

435 440 445

Leu Glu Glu Asp Gly Glu Ser Asp Thr Glu Lys Asp Ser Gly Ser Gln

450 455 460

Val Glu Glu Asp Thr Ser Cys Ser Asp Ser Lys Thr Leu Cys Thr Gln

465 470 475 480

Asp Ser Glu Ser Thr Glu Ile Pro Leu Asp Glu Gln Val Glu Glu Glu

485 490 495

Ala Val Gly Glu Glu Glu Glu Ser Gln Pro Glu Ala Cys Val Ile Asp

500 505 510

Asp Arg Ser Pro Asp Thr

515

<210> 19

<211> 8130

<212> DNA

<213> Homo sapiens

<400> 19

agattatagc ccagcgtacg agaagcacga gtcctatagt tggcgtaccc tgaggcctgc 60

cagttcctgc cttaatgcat atgtagtcgt aattgagttc tgacacggcc ttggatgttt 120

ctgtcctaaa tagctgacat tgcatcttca agactgtcat tccagttggc ttttgagtgg 180

atacgtgcag tgagatcatt gacactggaa acactagttc ccattttaat tacttaaaac 240

accacgatga aaagaaatac ctgtgatttg ctttctcgga gcaaaagtgc ctctgaggaa 300

acactacatt ccagtaatga agaggaagac ccttccgcg gaatggaacc ctatcttgtc 360

cggagacttt catgtcgcaa tattcagctt ccccctctcg ccttcagaca gttggaacaa 420

gctgacttga aaagtgaatc agagaacatt caacgaccaa ccagcctccc cctgaagatt 480

ctgccgctga ttgctatcac ttctgcagaa tccagtggtt ttgatgtgga caatggcaca 540

tctgcgggac ggagtccctt ggatcccatg accagcccag gatccggggct aattctccaa 600

gcaaattttg tccacagtca acgacgggag tccttcctgt atcgatccga cagcgattat 660

gacctctctc caaagtctat gtcccggaac tcctccattg ccagtgatat acacggagat 720

gacttgattg tgactccatt tgctcaggtc ttggccagtc tgcgaactgt acgaaacaac 780

tttgctgcat taactaattt gcaagatcga gcacctagca aaagatcacc catgtgcaac 840

caaccatcca tcaacaaagc caccataaca gaggaggcct accagaaact ggccagcgag 900

accctggagg agctggactg gtgtctggac cagctagaga ccctacagac caggcactcc 960

gtcagtgaga tggcctccaa caagtttaaa aggatgctta atcgggagct cacccatctc 1020

tctgaaatga gtcggtctgg aaatcaagtg tcagagttta tatcaaacac attcttagat 1080

aagcaacatg aagtggaaat tccttctcca actcagaagg aaaaggagaa aaagaaaaga 1140

ccaatgtctc agatcagtgg agtcaagaaa ttgatgcaca gctctagtct gactaattca 1200

agtatcccaa ggtttggagt taaaactgaa caagaagatg tccttgccaa ggaactagaa 1260

gatgtgaaca aatggggtct tcatgttttc agaatagcag agttgtctgg taaccggccc 1320

ttgactgtta tcatgcacac catttttcag gaacgggatt tattaaaaac atttaaaatt 1380

ccagtagata ctttaattac atatcttatg actctcgaag accattacca tgctgatgtg 1440

gcctatcaca acaatatcca tgctgcagat gttgtccagt ctactcatgt gctattatct 1500

1560

gcaatacatg atgtagatca tcctggtgtg tccaatcaat ttctgatcaa tacaaactct 1620

gaacttgcct tgatgtacaa tgattcctca gtcttagaga accatcattt ggctgtggggc 1680

tttaaattgc ttcaggaaga aaactgtgac attttccaga atttgaccaa aaaacaaaga 1740

caatctttaa ggaaaatggt cattgacatc gtacttgcaa cagatatgtc aaaacacatg 1800

aatctactgg ctgatttgaa gactatggtt gaaactaaga aagtgacaag ctctggagtt 1860

cttcttcttg ataattattc cgataggatt caggttcttc agaatatggt gcactgtgca 1920

gatctgagca acccaacaaa gcctctccag ctgtaccgcc agtggacgga ccggataatg 1980

gaggagttct tccgccaagg agaccgagag agggaacgtg gcatggagat aagccccatg 2040

tgtgacaagc acaatgcttc cgtggaaaaa tcacaggtgg gcttcataga ctatattgtt 2100

catcccctct gggagacatg ggcagacctc gtccaccctg acgccccagga tattttggac 2160

actttggagg acaatcgtga atggtaccag agcacaatcc ctcagagccc ctctcctgca 2220

cctgatgacc cagaggaggg ccggcagggt caaactgaga aattccagtt tgaactaact 2280

ttagaggaag atggtgagtc agacacggaa aaggacagtg gcagtcaagt ggaagaagac 2340

actagctgca gtgactccaa gactctttgt actcaagact cagagtctac tgaaattccc 2400

cttgatgaac aggttgaaga ggaggcagta ggggaagaag aggaaagcca gcctgaagcc 2460

2520

ttttttaagt agaaaaattg tttccaaagt gcatgtcaca tgccacaacc acggtcacac 2580

2640

gctcaggaat atcgtaacca gttttttcac ctccatgtca tccgagcaag gtggacatct 2700

tcacgaacag cgtttttaac aagatttcag cttggtagag ctgacaaagc agataaaatc 2760

2820

2880

gtgaacttaa tggacgaagc aacaaatatg tcaagaacag gacatagcac gaatctgtta 2940

ccagtaggag gaggatgagc cacagaaatt gcataatttt ctaatttcaa gtcttcctga 3000

tacatgactg aatagtgtgg ttcagtgagc tgcactgacc tctacatttt gtatgatatg 3060

3120

3180

caagttttca gtacaaatct gtgctacact ggataaaaat ctaatttatg aattttactt 3240

3300 gcaccttata gttcatagca attaactgat ttgtagtgat

3360

aagtctttat tatttacttt gcattttgtt tcactctttc cagataagca gagttgctct 3420

tcaccagtgt ttttcttcat gtgcaaagtg actatttgtt ctataatact tttatgtgtg 3480

ttatatcaaa tgtgtcttaa gcttcatgca aactcagtca tcagttcgtg ttgtctgaag 3540

caagtgggag atatataaat acccagtagc taaaatggtc agtctttttt agatgttttc 3600

ctacttagta tctcctaata acgttttgct gtgtcactag atgttcattt cacaagtgca 3660

tgtctttcta ataatccaca catttcatgc tctaataatc cacacatttc atgctcattt 3720

ttattgtttt tacagccagt tatagtaaga aaaaggtttt tccccttgtg ctgcttttata 3780

atttagcgtg tgtctgaacc ttatccatgt ttgctagatg aggtcttgtc aaatatatca 3840

ctaccattgt caccggtgaa aagaaacagg tagttaagtt agggttaaca ttcatttcaa 3900

ccacgaggtt gtatatcatg actagctttt actcttggtt tacagagaaa agttaaacag 3960

ccaactaggc agtttttaag aatattaaca atatattaac aaacaccaat acaactaatc 4020

ctatttggtt ttaatgattt caccatggga ttaagaacta tatcaggaac atccctgaga 4080

aacggtttta agtgtagcaa ctactcttcc ttaatggaca gccacataac gtgtaggaag 4140

tcctttatca cttatcctcg atccataagc atatcttgca gaggggaact acttctttaa 4200

acacatggag ggaaagaaga tgatgccact ggcaccagag ggttagtact gtgatgcatc 4260

ctaaaatatt tattatattg gtaaaaattc tggttaaata aaaaattaga gatcactctt 4320

ggctgatttc agcaccagga actgtattac agttttagag attaattcct agtgtttacc 4380

tgattatagc agttggcatc atggggcatt taattctgac tttatcccca cgtcagcctt 4440

aataaagtct tctttacctt ctctatgaag actttaaagc ccaaataatc atttttcaca 4500

ttgatattca agaattgaga tagatagaag ccaaagtggg tatctgacaa gtggaaaatc 4560

aaacgtttaa gaagaattac aactctgaaa agcatttata tgtggaactt ctcaaggagc 4620

ctcctgggga ctggaaagta agtcatcagc caggcaaatg actcatgctg aagagagtcc 4680

ccatttcagt cccctgagat ctagctgatg cttagatcct ttgaaataaa aattatgtct 4740

ttataactct gatcttttac ataaagcaga agaggaatca actagttaat tgcaaggttt 4800

ctactctgtt tcctctgtaa agatcagatg gtaatctttc aaataagaaa aaaataaaga 4860

4920

ttgtccctga gtgaagtcta gaaagaaagg taaagagtct agagtttatt cctctttcca 4980

aaacattctc attcctctcc tccctacact tagtatttcc cccacagagt gcctagaatc 5040

ttaataatga ataaaataaa aagcagcaat atgtcattaa caaatccaga cctgaaaggg 5100

taaagggttt ataactgcac taataaagag aggctcttttt ttttctcttcc agtttgttgg 5160

tttttaatgg taccgtgttg taaagatacc cactaatgga caatcaaatt gcagaaaagg 5220

ctcaatatcc aagagacagg gactaatgca ctgtacaatc tgcttatcct tgcccttctc 5280

tcttgccaaa gtgtgcttca gaaatatata ctgctttaaa aaagaataaa agaatatcct 5340

tttacaagtg gctttacatt tcctaaaatg ccataagaaa atgcaatatc tgggtactgt 5400

atggggaaaa aaatgtccaa gtttgtgtaa aaccagtgca tttcagcttg caagttactg 5460

aacacaataa tgctgtttta attttgtttt atatcagtta aaattcacaa taatgtagat 5520

agaacaaatt acagacaagg aaagaaaaaa cttgaatgaa atggatttta cagaaagctt 5580

tatgataatt tttgaatgca ttatttattt tttgtgccat gcattttttt tctcaccaaa 5640

tgaccttacc tgtaatacag tcttgtttgt ctgtttacaa ccatgtattt attgcaatgt 5700

acatactgta atgttaattg taaattatct gttcttatta aaacatcatc ccatgatggg 5760

atggtgttga tatatttgga aactcttggt gagagaatga atggtgtgta tacatactct 5820

gtacattttt cttttctcct gtaatatagt cttgtcacct tagagcttgt ttatggaaga 5880

ttcaagaaaa ctataaaata cttaaagata tataaattta aaaaaacata gctgcaggtc 5940

tttggtccca gggctgtgcc ttaactttaa ccaatatttt cttctgtttt gctgcatttg 6000

aaaggtaaca gtggagctag ggctgggcat tttacatcca ggcttttaat tgattagaat 6060

6120

6180

tttgccatct gttgatcagg aactacttca gctacttgca tttgattatt tccttttttt 6240

ttttttttaa ctcggaaaca caactgggga aatatattct ttcccagtga ttataaacaa 6300

tctttttctt tttttttaagt ccttttggct tctagagctc ataggaaaat ggacttgatt 6360

tgaaattgga gccagagttt actcgtgttg gttatctatt catcagcttc ctgacatgtt 6420

aagagaatac attaaagaga aaatactgtt ttttaatcct aaaatttttc ttcactaag 6480

ataaaccaaa tgtccttaca tatatgtaaa cccatctatt taaacgcaaa ggtggggttga 6540

tgtcagttta catagcagaa agcattcact atcctctaag atttgtttct gcaaaacttt 6600

cattgcttta gaattttaaa atttcacctt gtacaatggc cagcccctaa agcaggaaac 6660

atttataatg gattatatgg aaacatcctc ccagtacttg cccagccctt gaatcatgtg 6720

6780

tattttattt tttgacacaa actgtagatt ttagcagccc tggcccaaag gaatttgatt 6840

acttttgttt taaacagtac aaaggggaca ctataattac aaaaacatcc ttaactgatt 6900

tgagttgttt ttatttcttt ggatatattt tcagagtggt aaattgtgtg tgagaattac 6960

aaatgattat tcttttagtg gtttcttagc ctctcttaca gcccacgggg atagtactgt 7020

acatcaatac cttcatatga aatttttata tgcaatgaaa ataaaagcat gggttgattc 7080

tgcctattta tgactcaatc ttttacaaat aaaagattat tcattttaaa ttatagttca 7140

atcagcatgt ctcttaggat actgaacgtg gttgaaatga aaggatagtg acatcataag 7200

7260

taccaaaatt acttagatgg cctataagat taagcatggt gttttctaag caagctttga 7320

aaggggcctt ccatacttac ttaattgaat attctgggat attgaaaatt attcagatac 7380

ttgacaatta tttttggtta cctactccgc aaactacaaa gttttaagga ctcaacaata 7440

agttaatgag acacagtgtt tgctttcatg gagcttacag tctggagggg acaaaggctt 7500

aaacaatact catataatta tatatgtgat cagtacaatg aaggagctca gtggggtaaa 7560

taagcaggaa cctgaacttg atctgttccg gagggccaca gaaggcttcc ttgaggcctt 7620

7680

acaggaagag gaaggggatc cagacaagaa acatttgcaa agatcttgag gcataaatga 7740

gcttgagaca tctggagaaa ctgaggaaaa gtgagagagt aggcagggcc tggagccgca 7800

gagccattgc taaccatcct gtgtgagata tcccccattc tgtagcttta ttctcataac 7860

cctgctcaat tttctttata acacttctca cagatttata tacgtgtttg tttttgttat 7920

ctgtctctcc caccagacca cagctccatg agagcaaggt ctttgcttac caatatatca 7980

ctagcactta aaactatgcc tggtacacag taggttctta atatgtgttg aatatagcca 8040

tcaaattgat attggatata attcaatctg ataagatatt ttgagatatt aaagagtttt 8100

taacttgata ccataaaaaaaaaaaaaaaa 8130

<210> 20

<211> 748

<212> PROTEIN

<213> Homo sapiens

<400> 20

Met Lys Arg Asn Thr Cys Asp Leu Leu Ser Arg Ser Lys Ser Ala Ser

1 5 10 15

Glu Glu Thr Leu His Ser Ser Asn Glu Glu Glu Asp Pro Phe Arg Gly

20 25 30

Met Glu Pro Tyr Leu Val Arg Arg Leu Ser Cys Arg Asn Ile Gln Leu

35 40 45

Pro Pro Leu Ala Phe Arg Gln Leu Glu Gln Ala Asp Leu Lys Ser Glu

50 55 60

Ser Glu Asn Ile Gln Arg Pro Thr Ser Leu Pro Leu Lys Ile Leu Pro

65 70 75 80

Leu Ile Ala Ile Thr Ser Ala Glu Ser Ser Gly Phe Asp Val Asp Asn

85 90 95

Gly Thr Ser Ala Gly Arg Ser Pro Leu Asp Pro Met Thr Ser Pro Gly

100 105 110

Ser Gly Leu Ile Leu Gln Ala Asn Phe Val His Ser Gln Arg Arg Glu

115 120 125

Ser Phe Leu Tyr Arg Ser Asp Ser Asp Tyr Asp Leu Ser Pro Lys Ser

130 135 140

Met Ser Arg Asn Ser Ser Ile Ala Ser Asp Ile His Gly Asp Asp Leu

145 150 155 160

Ile Val Thr Pro Phe Ala Gln Val Leu Ala Ser Leu Arg Thr Val Arg

165 170 175

Asn Asn Phe Ala Ala Leu Thr Asn Leu Gln Asp Arg Ala Pro Ser Lys

180 185 190

Arg Ser Pro Met Cys Asn Gln Pro Ser Ile Asn Lys Ala Thr Ile Thr

195 200 205

Glu Glu Ala Tyr Gln Lys Leu Ala Ser Glu Thr Leu Glu Glu Glu Leu Asp

210 215 220

Trp Cys Leu Asp Gln Leu Glu Thr Leu Gln Thr Arg His Ser Val Ser

225 230 235 240

Glu Met Ala Ser Asn Lys Phe Lys Arg Met Leu Asn Arg Glu Leu Thr

245 250 255

His Leu Ser Glu Met Ser Arg Ser Gly Asn Gln Val Ser Glu Phe Ile

260 265 270

Ser Asn Thr Phe Leu Asp Lys Gln His Glu Val Glu Ile Pro Ser Pro

275 280 285

Thr Gln Lys Glu Lys Glu Lys Lys Lys Arg Pro Met Ser Gln Ile Ser

290 295 300

Gly Val Lys Lys Leu Met His Ser Ser Ser Leu Thr Asn Ser Ser Ile

305 310 315 320

Pro Arg Phe Gly Val Lys Thr Glu Gln Glu Asp Val Leu Ala Lys Glu

325 330 335

Leu Glu Asp Val Asn Lys Trp Gly Leu His Val Phe Arg Ile Ala Glu

340 345 350

Leu Ser Gly Asn Arg Pro Leu Thr Val Ile Met His Thr Ile Phe Gln

355 360 365

Glu Arg Asp Leu Leu Lys Thr Phe Lys Ile Pro Val Asp Thr Leu Ile

370 375 380

Thr Tyr Leu Met Thr Leu Glu Asp His Tyr His Ala Asp Val Ala Tyr

385 390 395 400

His Asn Asn Ile His Ala Ala Asp Val Val Gln Ser Thr His Val Leu

405 410 415

Leu Ser Thr Pro Ala Leu Glu Ala Val Phe Thr Asp Leu Glu Ile Leu

420 425 430

Ala Ala Ile Phe Ala Ser Ala Ile His Asp Val Asp His Pro Gly Val

435 440 445

Ser Asn Gln Phe Leu Ile Asn Thr Asn Ser Glu Leu Ala Leu Met Tyr

450 455 460

Asn Asp Ser Ser Val Leu Glu Asn His His Leu Ala Val Gly Phe Lys

465 470 475 480

Leu Leu Gln Glu Glu Asn Cys Asp Ile Phe Gln Asn Leu Thr Lys Lys

485 490 495

Gln Arg Gln Ser Leu Arg Lys Met Val Ile Asp Ile Val Leu Ala Thr

500 505 510

Asp Met Ser Lys His Met Asn Leu Leu Ala Asp Leu Lys Thr Met Val

515 520 525

Glu Thr Lys Lys Val Thr Ser Ser Gly Val Leu Leu Leu Asp Asn Tyr

530 535 540

Ser Asp Arg Ile Gln Val Leu Gln Asn Met Val His Cys Ala Asp Leu

545 550 555 560

Ser Asn Pro Thr Lys Pro Leu Gln Leu Tyr Arg Gln Trp Thr Asp Arg

565 570 575

Ile Met Glu Glu Phe Phe Arg Gln Gly Asp Arg Glu Arg Glu Arg Gly

580 585 590

Met Glu Ile Ser Pro Met Cys Asp Lys His Asn Ala Ser Val Glu Lys

595 600 605

Ser Gln Val Gly Phe Ile Asp Tyr Ile Val His Pro Leu Trp Glu Thr

610 615 620

Trp Ala Asp Leu Val His Pro Asp Ala Gln Asp Ile Leu Asp Thr Leu

625 630 635 640

Glu Asp Asn Arg Glu Trp Tyr Gln Ser Thr Ile Pro Gln Ser Pro Ser

645 650 655

Pro Ala Pro Asp Asp Pro Glu Glu Gly Arg Gln Gly Gln Thr Glu Lys

660 665 670

Phe Gln Phe Glu Leu Thr Leu Glu Glu Asp Gly Glu Ser Asp Thr Glu

675 680 685

Lys Asp Ser Gly Ser Gln Val Glu Glu Asp Thr Ser Cys Ser Asp Ser

690 695 700

Lys Thr Leu Cys Thr Gln Asp Ser Glu Ser Thr Glu Ile Pro Leu Asp

705 710 715 720

Glu Gln Val Glu Glu Glu Ala Val Gly Glu Glu Glu Glu Ser Gln Pro

725 730 735

Glu Ala Cys Val Ile Asp Asp Arg Ser Pro Asp Thr

740 745

<210> 21

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> PDE4D7_forward primer

<400> 21

gaacattcaa cgaccaacca 20

<210> 22

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> PDE4D7_reverse primer

<400> 22

tgccattgtc cacatcaaaa 20

<210> 23

<211> 27

<212> DNA

<213> Artificial sequence

<220>

<223> PDE4D7 probe

<400> 23

ctgccgctga ttgctatcac ttctgca 27

<210> 24

<211> 22

<212> DNA

<213> Artificial sequence

<220>

<223> PDE4D7 forward primer 2

<400> 24

cgctgattgc tatcacttct gc 22

<210> 25

<211> 24

<212> DNA

<213> Artificial sequence

<220>

<223> PDE4D7 reverse primer

<400> 25

gtcgttgact gtggacaaaa tttg 24

<210> 26

<211> 34

<212> DNA

<213> Artificial sequence

<220>

<223> PDE4D7 probe 2

<400> 26

ttcccttgga tcccatgacc agcccataag ggaa 34

<210> 27

<211> 8130

<212> DNA

<213> Homo sapiens

<400> 27

agattatagc ccagcgtacg agaagcacga gtcctatagt tggcgtaccc tgaggcctgc 60

cagttcctgc cttaatgcat atgtagtcgt aattgagttc tgacacggcc ttggatgttt 120

ctgtcctaaa tagctgacat tgcatcttca agactgtcat tccagttggc ttttgagtgg 180

atacgtgcag tgagatcatt gacactggaa acactagttc ccattttaat tacttaaaac 240

accacgatga aaagaaatac ctgtgatttg ctttctcgga gcaaaagtgc ctctgaggaa 300

acactacatt ccagtaatga agaggaagac ccttccgcg gaatggaacc ctatcttgtc 360

cggagacttt catgtcgcaa tattcagctt ccccctctcg ccttcagaca gttggaacaa 420

gctgacttga aaagtgaatc agagaacatt caacgaccaa ccagcctccc cctgaagatt 480

ctgccgctga ttgctatcac ttctgcagaa tccagtggtt ttgatgtgga caatggcaca 540

tctgcgggac ggagtccctt ggatcccatg accagcccag gatccggggct aattctccaa 600

gcaaattttg tccacagtca acgacgggag tccttcctgt atcgatccga cagcgattat 660

gacctctctc caaagtctat gtcccggaac tcctccattg ccagtgatat acacggagat 720

gacttgattg tgactccatt tgctcaggtc ttggccagtc tgcgaactgt acgaaacaac 780

tttgctgcat taactaattt gcaagatcga gcacctagca aaagatcacc catgtgcaac 840

caaccatcca tcaacaaagc caccataaca gaggaggcct accagaaact ggccagcgag 900

accctggagg agctggactg gtgtctggac cagctagaga ccctacagac caggcactcc 960

gtcagtgaga tggcctccaa caagtttaaa aggatgctta atcgggagct cacccatctc 1020

tctgaaatga gtcggtctgg aaatcaagtg tcagagttta tatcaaacac attcttagat 1080

aagcaacatg aagtggaaat tccttctcca actcagaagg aaaaggagaa aaagaaaaga 1140

ccaatgtctc agatcagtgg agtcaagaaa ttgatgcaca gctctagtct gactaattca 1200

agtatcccaa ggtttggagt taaaactgaa caagaagatg tccttgccaa ggaactagaa 1260

gatgtgaaca aatggggtct tcatgttttc agaatagcag agttgtctgg taaccggccc 1320

ttgactgtta tcatgcacac catttttcag gaacgggatt tattaaaaac atttaaaatt 1380

ccagtagata ctttaattac atatcttatg actctcgaag accattacca tgctgatgtg 1440

gcctatcaca acaatatcca tgctgcagat gttgtccagt ctactcatgt gctattatct 1500

1560

gcaatacatg atgtagatca tcctggtgtg tccaatcaat ttctgatcaa tacaaactct 1620

gaacttgcct tgatgtacaa tgattcctca gtcttagaga accatcattt ggctgtggggc 1680

tttaaattgc ttcaggaaga aaactgtgac attttccaga atttgaccaa aaaacaaaga 1740

caatctttaa ggaaaatggt cattgacatc gtacttgcaa cagatatgtc aaaacacatg 1800

aatctactgg ctgatttgaa gactatggtt gaaactaaga aagtgacaag ctctggagtt 1860

cttcttcttg ataattattc cgataggatt caggttcttc agaatatggt gcactgtgca 1920

gatctgagca acccaacaaa gcctctccag ctgtaccgcc agtggacgga ccggataatg 1980

gaggagttct tccgccaagg agaccgagag agggaacgtg gcatggagat aagccccatg 2040

tgtgacaagc acaatgcttc cgtggaaaaa tcacaggtgg gcttcataga ctatattgtt 2100

catcccctct gggagacatg ggcagacctc gtccaccctg acgccccagga tattttggac 2160

actttggagg acaatcgtga atggtaccag agcacaatcc ctcagagccc ctctcctgca 2220

cctgatgacc cagaggaggg ccggcagggt caaactgaga aattccagtt tgaactaact 2280

ttagaggaag atggtgagtc agacacggaa aaggacagtg gcagtcaagt ggaagaagac 2340

actagctgca gtgactccaa gactctttgt actcaagact cagagtctac tgaaattccc 2400

cttgatgaac aggttgaaga ggaggcagta ggggaagaag aggaaagcca gcctgaagcc 2460

2520

ttttttaagt agaaaaattg tttccaaagt gcatgtcaca tgccacaacc acggtcacac 2580

2640

gctcaggaat atcgtaacca gttttttcac ctccatgtca tccgagcaag gtggacatct 2700

tcacgaacag cgtttttaac aagatttcag cttggtagag ctgacaaagc agataaaatc 2760

2820

2880

gtgaacttaa tggacgaagc aacaaatatg tcaagaacag gacatagcac gaatctgtta 2940

ccagtaggag gaggatgagc cacagaaatt gcataatttt ctaatttcaa gtcttcctga 3000

tacatgactg aatagtgtgg ttcagtgagc tgcactgacc tctacatttt gtatgatatg 3060

3120

3180

caagttttca gtacaaatct gtgctacact ggataaaaat ctaatttatg aattttactt 3240

3300 gcaccttata gttcatagca attaactgat ttgtagtgat

3360

aagtctttat tatttacttt gcattttgtt tcactctttc cagataagca gagttgctct 3420

tcaccagtgt ttttcttcat gtgcaaagtg actatttgtt ctataatact tttatgtgtg 3480

ttatatcaaa tgtgtcttaa gcttcatgca aactcagtca tcagttcgtg ttgtctgaag 3540

caagtgggag atatataaat acccagtagc taaaatggtc agtctttttt agatgttttc 3600

ctacttagta tctcctaata acgttttgct gtgtcactag atgttcattt cacaagtgca 3660

tgtctttcta ataatccaca catttcatgc tctaataatc cacacatttc atgctcattt 3720

ttattgtttt tacagccagt tatagtaaga aaaaggtttt tccccttgtg ctgcttttata 3780

atttagcgtg tgtctgaacc ttatccatgt ttgctagatg aggtcttgtc aaatatatca 3840

ctaccattgt caccggtgaa aagaaacagg tagttaagtt agggttaaca ttcatttcaa 3900

ccacgaggtt gtatatcatg actagctttt actcttggtt tacagagaaa agttaaacag 3960

ccaactaggc agtttttaag aatattaaca atatattaac aaacaccaat acaactaatc 4020

ctatttggtt ttaatgattt caccatggga ttaagaacta tatcaggaac atccctgaga 4080

aacggtttta agtgtagcaa ctactcttcc ttaatggaca gccacataac gtgtaggaag 4140

tcctttatca cttatcctcg atccataagc atatcttgca gaggggaact acttctttaa 4200

acacatggag ggaaagaaga tgatgccact ggcaccagag ggttagtact gtgatgcatc 4260

ctaaaatatt tattatattg gtaaaaattc tggttaaata aaaaattaga gatcactctt 4320

ggctgatttc agcaccagga actgtattac agttttagag attaattcct agtgtttacc 4380

tgattatagc agttggcatc atggggcatt taattctgac tttatcccca cgtcagcctt 4440

aataaagtct tctttacctt ctctatgaag actttaaagc ccaaataatc atttttcaca 4500

ttgatattca agaattgaga tagatagaag ccaaagtggg tatctgacaa gtggaaaatc 4560

aaacgtttaa gaagaattac aactctgaaa agcatttata tgtggaactt ctcaaggagc 4620

ctcctgggga ctggaaagta agtcatcagc caggcaaatg actcatgctg aagagagtcc 4680

ccatttcagt cccctgagat ctagctgatg cttagatcct ttgaaataaa aattatgtct 4740

ttataactct gatcttttac ataaagcaga agaggaatca actagttaat tgcaaggttt 4800

ctactctgtt tcctctgtaa agatcagatg gtaatctttc aaataagaaa aaaataaaga 4860

4920

ttgtccctga gtgaagtcta gaaagaaagg taaagagtct agagtttatt cctctttcca 4980

aaacattctc attcctctcc tccctacact tagtatttcc cccacagagt gcctagaatc 5040

ttaataatga ataaaataaa aagcagcaat atgtcattaa caaatccaga cctgaaaggg 5100

taaagggttt ataactgcac taataaagag aggctcttttt ttttctcttcc agtttgttgg 5160

tttttaatgg taccgtgttg taaagatacc cactaatgga caatcaaatt gcagaaaagg 5220

ctcaatatcc aagagacagg gactaatgca ctgtacaatc tgcttatcct tgcccttctc 5280

tcttgccaaa gtgtgcttca gaaatatata ctgctttaaa aaagaataaa agaatatcct 5340

tttacaagtg gctttacatt tcctaaaatg ccataagaaa atgcaatatc tgggtactgt 5400

atggggaaaa aaatgtccaa gtttgtgtaa aaccagtgca tttcagcttg caagttactg 5460

aacacaataa tgctgtttta attttgtttt atatcagtta aaattcacaa taatgtagat 5520

agaacaaatt acagacaagg aaagaaaaaa cttgaatgaa atggatttta cagaaagctt 5580

tatgataatt tttgaatgca ttatttattt tttgtgccat gcattttttt tctcaccaaa 5640

tgaccttacc tgtaatacag tcttgtttgt ctgtttacaa ccatgtattt attgcaatgt 5700

acatactgta atgttaattg taaattatct gttcttatta aaacatcatc ccatgatggg 5760

atggtgttga tatatttgga aactcttggt gagagaatga atggtgtgta tacatactct 5820

gtacattttt cttttctcct gtaatatagt cttgtcacct tagagcttgt ttatggaaga 5880

ttcaagaaaa ctataaaata cttaaagata tataaattta aaaaaacata gctgcaggtc 5940

tttggtccca gggctgtgcc ttaactttaa ccaatatttt cttctgtttt gctgcatttg 6000

aaaggtaaca gtggagctag ggctgggcat tttacatcca ggcttttaat tgattagaat 6060

6120

6180

tttgccatct gttgatcagg aactacttca gctacttgca tttgattatt tccttttttt 6240

ttttttttaa ctcggaaaca caactgggga aatatattct ttcccagtga ttataaacaa 6300

tctttttctt tttttttaagt ccttttggct tctagagctc ataggaaaat ggacttgatt 6360

tgaaattgga gccagagttt actcgtgttg gttatctatt catcagcttc ctgacatgtt 6420

aagagaatac attaaagaga aaatactgtt ttttaatcct aaaatttttc ttcactaag 6480

ataaaccaaa tgtccttaca tatatgtaaa cccatctatt taaacgcaaa ggtggggttga 6540

tgtcagttta catagcagaa agcattcact atcctctaag atttgtttct gcaaaacttt 6600

cattgcttta gaattttaaa atttcacctt gtacaatggc cagcccctaa agcaggaaac 6660

atttataatg gattatatgg aaacatcctc ccagtacttg cccagccctt gaatcatgtg 6720

6780

tattttattt tttgacacaa actgtagatt ttagcagccc tggcccaaag gaatttgatt 6840

acttttgttt taaacagtac aaaggggaca ctataattac aaaaacatcc ttaactgatt 6900

tgagttgttt ttatttcttt ggatatattt tcagagtggt aaattgtgtg tgagaattac 6960

aaatgattat tcttttagtg gtttcttagc ctctcttaca gcccacgggg atagtactgt 7020

acatcaatac cttcatatga aatttttata tgcaatgaaa ataaaagcat gggttgattc 7080

tgcctattta tgactcaatc ttttacaaat aaaagattat tcattttaaa ttatagttca 7140

atcagcatgt ctcttaggat actgaacgtg gttgaaatga aaggatagtg acatcataag 7200

7260

taccaaaatt acttagatgg cctataagat taagcatggt gttttctaag caagctttga 7320

aaggggcctt ccatacttac ttaattgaat attctgggat attgaaaatt attcagatac 7380

ttgacaatta tttttggtta cctactccgc aaactacaaa gttttaagga ctcaacaata 7440

agttaatgag acacagtgtt tgctttcatg gagcttacag tctggagggg acaaaggctt 7500

aaacaatact catataatta tatatgtgat cagtacaatg aaggagctca gtggggtaaa 7560

taagcaggaa cctgaacttg atctgttccg gagggccaca gaaggcttcc ttgaggcctt 7620

7680

acaggaagag gaaggggatc cagacaagaa acatttgcaa agatcttgag gcataaatga 7740

gcttgagaca tctggagaaa ctgaggaaaa gtgagagagt aggcagggcc tggagccgca 7800

gagccattgc taaccatcct gtgtgagata tcccccattc tgtagcttta ttctcataac 7860

cctgctcaat tttctttata acacttctca cagatttata tacgtgtttg tttttgttat 7920

ctgtctctcc caccagacca cagctccatg agagcaaggt ctttgcttac caatatatca 7980

ctagcactta aaactatgcc tggtacacag taggttctta atatgtgttg aatatagcca 8040

tcaaattgat attggatata attcaatctg ataagatatt ttgagatatt aaagagtttt 8100

taacttgata ccataaaaaaaaaaaaaaaa 8130

<210> 28

<211> 687

<212> PROTEIN

<213> Homo sapiens

<400> 28

Met Ala Phe Val Trp Asp Pro Leu Gly Ala Thr Val Pro Gly Pro Ser

1 5 10 15

Thr Arg Ala Lys Ser Arg Leu Arg Phe Ser Lys Ser Tyr Ser Phe Asp

20 25 30

Val Asp Asn Gly Thr Ser Ala Gly Arg Ser Pro Leu Asp Pro Met Thr

35 40 45

Ser Pro Gly Ser Gly Leu Ile Leu Gln Ala Asn Phe Val His Ser Gln

50 55 60

Arg Arg Glu Ser Phe Leu Tyr Arg Ser Asp Ser Asp Tyr Asp Leu Ser

65 70 75 80

Pro Lys Ser Met Ser Arg Asn Ser Ser Ile Ala Ser Asp Ile His Gly

85 90 95

Asp Asp Leu Ile Val Thr Pro Phe Ala Gln Val Leu Ala Ser Leu Arg

100 105 110

Thr Val Arg Asn Asn Phe Ala Ala Leu Thr Asn Leu Gln Asp Arg Ala

115 120 125

Pro Ser Lys Arg Ser Pro Met Cys Asn Gln Pro Ser Ile Asn Lys Ala

130 135 140

Thr Ile Thr Glu Glu Ala Tyr Gln Lys Leu Ala Ser Glu Thr Leu Glu

145 150 155 160

Glu Leu Asp Trp Cys Leu Asp Gln Leu Glu Thr Leu Gln Thr Arg His

165 170 175

Ser Val Ser Glu Met Ala Ser Asn Lys Phe Lys Arg Met Leu Asn Arg

180 185 190

Glu Leu Thr His Leu Ser Glu Met Ser Arg Ser Gly Asn Gln Val Ser

195 200 205

Glu Phe Ile Ser Asn Thr Phe Leu Asp Lys Gln His Glu Val Glu Ile

210 215 220

Pro Ser Pro Thr Gln Lys Glu Lys Glu Lys Lys Lys Arg Pro Met Ser

225 230 235 240

Gln Ile Ser Gly Val Lys Lys Leu Met His Ser Ser Ser Leu Thr Asn

245 250 255

Ser Ser Ile Pro Arg Phe Gly Val Lys Thr Glu Gln Glu Asp Val Leu

260 265 270

Ala Lys Glu Leu Glu Asp Val Asn Lys Trp Gly Leu His Val Phe Arg

275 280 285

Ile Ala Glu Leu Ser Gly Asn Arg Pro Leu Thr Val Ile Met His Thr

290 295 300

Ile Phe Gln Glu Arg Asp Leu Leu Lys Thr Phe Lys Ile Pro Val Asp

305 310 315 320

Thr Leu Ile Thr Tyr Leu Met Thr Leu Glu Asp His Tyr His Ala Asp

325 330 335

Val Ala Tyr His Asn Asn Ile His Ala Ala Asp Val Val Gln Ser Thr

340 345 350

His Val Leu Leu Ser Thr Pro Ala Leu Glu Ala Val Phe Thr Asp Leu

355 360 365

Glu Ile Leu Ala Ala Ile Phe Ala Ser Ala Ile His Asp Val Asp His

370 375 380

Pro Gly Val Ser Asn Gln Phe Leu Ile Asn Thr Asn Ser Glu Leu Ala

385 390 395 400

Leu Met Tyr Asn Asp Ser Ser Val Leu Glu Asn His His Leu Ala Val

405 410 415

Gly Phe Lys Leu Leu Gln Glu Glu Asn Cys Asp Ile Phe Gln Asn Leu

420 425 430

Thr Lys Lys Gln Arg Gln Ser Leu Arg Lys Met Val Ile Asp Ile Val

435 440 445

Leu Ala Thr Asp Met Ser Lys His Met Asn Leu Leu Ala Asp Leu Lys

450 455 460

Thr Met Val Glu Thr Lys Lys Val Thr Ser Ser Gly Val Leu Leu Leu

465 470 475 480

Asp Asn Tyr Ser Asp Arg Ile Gln Val Leu Gln Asn Met Val His Cys

485 490 495

Ala Asp Leu Ser Asn Pro Thr Lys Pro Leu Gln Leu Tyr Arg Gln Trp

500 505 510

Thr Asp Arg Ile Met Glu Glu Phe Phe Arg Gln Gly Asp Arg Glu Arg

515 520 525

Glu Arg Gly Met Glu Ile Ser Pro Met Cys Asp Lys His Asn Ala Ser

530 535 540

Val Glu Lys Ser Gln Val Gly Phe Ile Asp Tyr Ile Val His Pro Leu

545 550 555 560

Trp Glu Thr Trp Ala Asp Leu Val His Pro Asp Ala Gln Asp Ile Leu

565 570 575

Asp Thr Leu Glu Asp Asn Arg Glu Trp Tyr Gln Ser Thr Ile Pro Gln

580 585 590

Ser Pro Ser Pro Ala Pro Asp Asp Pro Glu Glu Gly Arg Gln Gly Gln

595 600 605

Thr Glu Lys Phe Gln Phe Glu Leu Thr Leu Glu Glu Asp Gly Glu Ser

610 615 620

Asp Thr Glu Lys Asp Ser Gly Ser Gln Val Glu Glu Asp Thr Ser Cys

625 630 635 640

Ser Asp Ser Lys Thr Leu Cys Thr Gln Asp Ser Glu Ser Thr Glu Ile

645 650 655

Pro Leu Asp Glu Gln Val Glu Glu Glu Ala Val Gly Glu Glu Glu Glu

660 665 670

Ser Gln Pro Glu Ala Cys Val Ile Asp Asp Arg Ser Pro Asp Thr

675 680 685

<210> 29

<211> 8395

<212> DNA

<213> Homo sapiens

<400> 29

ttctcactgc cctgcggtgt tttgaactgc cttcttacag acgtcataca gcccttgagg 60

aatagtttct gcctggtgag attgaatgat agttctcatt cacaaaaccc tggattctaa 120

gcagggacac acagaaatta ctttcgcagg taaatcagcc cacccagcca aagtgtggag 180

agatttgttc cttggctgac ttctttgctc cacggagagg agtgttttcc tgtgcttgcc 240

ctgaaatgga acttccttga cagctctccc gtgttacagt acctcccggt cattttcttt 300

ttctctctct ctacctgcgc tcttcgagtg tcagaaacct ttaaagctgt tactatggaa 360

ttgcaaaaaa gagatcaagt gactctttca ctatgctggt ttcccttgtg acccagatga 420

agaatcaatt cagaattcag ttcctccctt ggcattgcaa gacacagaag aaactgtcac 480

ttcctaacag cctagtactg gagtaaattc agtatgaagg aagaaagcgc tcctgcgtgt 540

tagaaccttg cccatgagct ggaccgagga caggagatgg actccaggaa aattggattt 600

660

tagaatgtat taatcagaat agttgaagac ttttttcct ttttattttt tttcaaaatg 720

agcattatta tgaagccaag atcccgatct acaagttccc taaggactgc agaggcagtt 780

tgttttgatg tggacaatgg cacatctgcg ggacggagtc ccttggatcc catgaccagc 840

ccaggatccg ggctaattct ccaagcaaat tttgtccaca gtcaacgacg ggagtccttc 900

ctgtatcgat ccgacagcga ttatgacctc tctccaaagt ctatgtcccg gaactcctcc 960

attgccagtg atatacacgg agatgacttg attgtgactc catttgctca ggtcttggcc 1020

agtctgcgaa ctgtacgaaa caactttgct gcattaacta atttgcaaga tcgagcacct 1080

agcaaaagat cacccatgtg caaccaacca tccatcaaca aagccaccat aacagaggag 1140

gcctaccaga aactggccag cgagaccctg gaggagctgg actggtgtct ggaccagcta 1200

1260

cttaatcggg agctcaccca tctctctgaa atgagtcggt ctggaaatca agtgtcagag 1320

tttatatcaa acacattctt agataagcaa catgaagtgg aaattccttc tccaactcag 1380

aaggaaaagg agaaaaagaa aagaccaatg tctcagatca gtggagtcaa gaaattgatg 1440

cacagctcta gtctgactaa ttcaagtatc ccaaggtttg gagttaaaac tgaacaagaa 1500

1560

gcagagttgt ctggtaaccg gcccttgact gttatcatgc acaccatttt tcaggaacgg 1620

gatttattaa aaacatttaa aattccagta gatactttaa ttacatatct tatgactctc 1680

gaagaccatt accatgctga tgtggcctat cacaacaata tccatgctgc agatgttgtc 1740

cagtctactc atgtgctatt atctacacct gctttggagg ctgtgtttac agatttggag 1800

attcttgcag caatttttgc cagtgcaata catgatgtag atcatcctgg tgtgtccaat 1860

caatttctga tcaatacaaa ctctgaactt gccttgatgt acaatgattc ctcagtctta 1920

gagaaccatc atttggctgt gggctttaaa ttgcttcagg aagaaaactg tgacattttc 1980

cagaatttga ccaaaaaaca aagacaatct ttaaggaaaa tggtcattga catcgtactt 2040

gcaacagata tgtcaaaaca catgaatcta ctggctgatt tgaagactat ggttgaaact 2100

aagaaagtga caagctctgg agttcttctt cttgataatt attccgatag gattcaggtt 2160

cttcagaata tggtgcactg tgcagatctg agcaacccaa caaagcctct ccagctgtac 2220

2280

cgtggcatgg agataagccc catgtgtgac aagcacaatg cttccgtgga aaaatcacag 2340

gtgggcttca tagactatat tgttcatccc ctctgggaga catgggcaga cctcgtccac 2400

cctgacgccc aggatatttt ggacactttg gaggacaatc gtgaatggta ccagagcaca 2460

atccctcaga gcccctctcc tgcacctgat gacccagagg agggccggca gggtcaaact 2520

gagaaattcc agtttgaact aactttagag gaagatggtg agtcagacac ggaaaaggac 2580

2640

gactcagagt ctactgaaat tccccttgat gaacaggttg aagagggaggc agtaggggaa 2700

gaagaggaaa gccagcctga agcctgtgtc atagatgatc gttctcctga cacgtaacag 2760

tgcaaaaact ttcatgcctt tttttttttt aagtagaaaa attgtttcca aagtgcatgt 2820

cacatgccac aaccacggtc acacctcact gtcatctgcc aggacgtttg ttgaacaaaa 2880

ctgaccttga ctactcagtc cagcgctcag gaatatcgta accagttttt tcacctccat 2940

gtcatccgag caaggtggac atcttcacga acagcgtttt taacaagatt tcagcttggt 3000

agagctgaca aagcagataa aatctactcc aaattatttt caagagagtg tgactcatca 3060

ggcagcccaa aagtttattg gacttggggt ttctattcct ttttatttgt ttgcaatatt 3120

ttcagaaga aggcattgca cagagtgaac ttaatggacg aagcaacaaa tatgtcaaga 3180

acaggacata gcacgaatct gttaccagta ggaggaggat gagccacaga aattgcataa 3240

ttttctaatt tcaagtcttc ctgatacatg actgaatagt gtggttcagt gagctgcact 3300

3360

aaatgtattg aggtattata tttaaaaaaa actatgttca gaacttcatc tgccactggt 3420

tatttttttc taaggagtaa cttgcaagtt ttcagtacaa atctgtgcta cactggataa 3480

3540

3600

3660

3720

3780

gtcatcagtt cgtgttgtct gaagcaagtg ggagatatat aaatacccag tagctaaaat 3840

3900

ctagatgttc atttcacaag tgcatgtctt tctaataatc cacacatttc atgctctaat 3960

aatccacaca tttcatgctc atttttattg tttttacagc cagttatagt aagaaaaagg 4020

tttttcccct tgtgctgctt tataatttag cgtgtgtctg aaccttatcc atgtttgcta 4080

gatgaggtct tgtcaaatat atcactacca ttgtcaccgg tgaaaagaaa caggtagtta 4140

agttagggtt aacattcatt tcaaccacga ggttgtatat catgactagc ttttactctt 4200

ggtttacaga gaaaagttaa acagccaact aggcagtttt taagaatatt aacaatatat 4260

taacaaacac caatacaact aatcctattt ggttttaatg atttcaccat gggattaaga 4320

actatatcag gaacatccct gagaaacggt tttaagtgta gcaactactc ttccttaatg 4380

gacagccaca taacgtgtag gaagtccttt atcacttatc ctcgatccat aagcatatct 4440

tgcagagggg aactacttct ttaaacacat ggagggaaag aagatgatgc cactggcacc 4500

agagggttag tactgtgatg catcctaaaa tatttattat attggtaaaa attctggtta 4560

aataaaaaat tagagatcac tcttggctga tttcagcacc aggaactgta ttacagtttt 4620

agagattaat tcctagtgtt tacctgatta tagcagttgg catcatgggg catttaattc 4680

tgactttatc cccacgtcag ccttaataaa gtcttcttta ccttctctat gaagacttta 4740

aagcccaaat aatcattttt cacattgata ttcaagaatt gagatagata gaagccaaag 4800

tgggtatctg acaagtggaa aatcaaacgt ttaagaagaa ttacaactct gaaaagcatt 4860

tatatgtgga acttctcaag gagcctcctg gggactggaa agtaagtcat cagccaggca 4920

aatgactcat gctgaagaga gtccccattt cagtcccctg agatctagct gatgcttaga 4980

tcctttgaaa taaaaattat gtctttataa ctctgatctt ttacataaag cagaagagga 5040

atcaactagt taattgcaag gtttctactc tgtttcctct gtaaagatca gatggtaatc 5100

tttcaaataa gaaaaaaata aagacgtatg

ggaacttgtt tcttttaatt ttatttgtcc ctgagtgaag tctagaaaga aaggtaaaga 5220

gtctagagtt tattcctctt tccaaaacat tctcattcct ctcctcccta cacttagtat 5280

ttcccccaca gagtgcctag aatcttaata atgaataaaa taaaaagcag caatatgtca 5340

ttaacaaatc cagacctgaa agggtaaagg gtttataact gcactaataa agagaggctc 5400

tttttttttc ttccagtttg ttggttttta atggtaccgt gttgtaaaga taccactaa 5460

tggacaatca aattgcagaa aaggctcaat atccaagaga cagggactaa tgcactgtac 5520

aatctgctta tccttgccct tctctcttgc caaagtgtgc ttcagaaata tatactgctt 5580

taaaaaagaa taaaagaata tccttttaca agtggcttta catttcctaa aatgccataa 5640

gaaaatgcaa tatctgggta ctgtatgggg aaaaaaatgt ccaagtttgt gtaaaaccag 5700

tgcatttcag cttgcaagtt actgaacaca ataatgctgt tttaattttg ttttatatca 5760

gttaaaattc acaataatgt agatagaaca aattacagac aaggaaagaa aaaacttgaa 5820

5880

ccatgcattt tttttctcac caaatgacct tacctgtaat acagtcttgt ttgtctgttt 5940

acaaccatgt atttattgca atgtacatac tgtaatgtta attgtaaatt atctgttctt 6000

attaaaacat catcccatga tgggatggtg ttgatatatt tggaaactct tggtgagaga 6060

atgaatggtg tgtatacata ctctgtacat ttttcttttc tcctgtaata tagtcttgtc 6120

6180

tttaaaaaaa catagctgca ggtctttggt cccagggctg tgccttaact ttaaccaata 6240

ttttcttctg ttttgctgca tttgaaaggt aacagtggag ctagggctgg gcattttaca 6300

6360

cctctgaaag attctgagac ccttttgaga cagaagctct taagtacttc ttgccaggga 6420

gcagcactgc atgtgtgatg gttgtttgcc atctgttgat caggaactac ttcagctact 6480

6540

ttctttccca gtgattataa acaatctttt tcttttttttt aagtcctttt ggcttctaga 6600

gctcatagga aaatggactt gatttgaaat tggagccaga gtttactcgt gttggttatc 6660

6720

6780

tatttaaacg caaaggtggg ttgatgtcag tttacatagc agaaagcatt cactatcctc 6840

6900

tggccagccc ctaaagcagg aaacatttat aatggattat atggaaacat cctcccagta 6960

cttgcccagc ccttgaatca tgtggctttt cagtgaaagg aaagattctt tttctaggaa 7020

aaatgagcct attttatttt attttatttt attttttgac acaaactgta gattttagca 7080

gccctggccc aaaggaattt gattactttt gttttaaaca gtacaaaggg gacactataa 7140

ttacaaaaac atccttaact gatttgagtt gtttttattt ctttggatat attttcagag 7200

tggtaaattg tgtgtgagaa ttacaaatga ttattctttt agtggtttct tagcctctct 7260

tacagcccac ggggatagta ctgtacatca ataccttcat atgaaatttt tatatgcaat 7320

gaaaataaaa gcatgggttg attctgccta tttatgactc aatcttttac aaataaaaga 7380

ttattcattt taaattatag ttcaatcagc atgtctctta ggatactgaa cgtggttgaa 7440

atgaaaggat agtgacatca taagttagta ctgatattca taaccaaata aagccaactt 7500

gagtaatttt gctacattaa aaattaccaa aattacttag atggcctata agattaagca 7560

tggtgttttc taagcaagct ttgaaagggg ccttccatac ttacttaatt gaatattctg 7620

ggatattgaa aattattcag atacttgaca attatttttg gttacctact ccgcaaacta 7680

catggagctt 7740

acagtctgga ggggacaaag gcttaaacaa tactcatata attatatatg tgatcagtac 7800

aatgaaggag ctcagtgggg taaataagca ggaacctgaa cttgatctgt tccggagggc 7860

cacagaaggc ttccttgagg ccttgagaaa gtgatttgca tctgagttct gaaggattgt 7920

aagaggtaac tagggaaaaa gttgacagga agaggaaggg gatccagaca agaaacattt 7980

gcaaagatct tgaggcataa atgagcttga gacatctgga gaaactgagg aaaagtgaga 8040

gagtaggcag ggcctggagc cgcagagcca ttgctaacca tcctgtgtga gatatccccc 8100

tataacactt ctcacagatt 8160

tatatacgtg tttgtttttg ttatctgtct ctcccaccag accacagctc catgagagca 8220

aggtctttgc ttaccaatat atcactagca cttaaaacta tgcctggtac acagtaggtt 8280

cttaattgt gttgaatata gccatcaaat tgatattgga tataattcaa tctgataaga 8340

tattttgaga tattaaagag tttttaactt gataccataa aaaaaaaaaa aaaaa 8395

<210> 30

<211> 679

<212> PROTEIN

<213> Homo sapiens

<400> 30

Met Ser Ile Ile Met Lys Pro Arg Ser Arg Ser Thr Ser Ser Leu Arg

1 5 10 15

Thr Ala Glu Ala Val Cys Phe Asp Val Asp Asn Gly Thr Ser Ala Gly

20 25 30

Arg Ser Pro Leu Asp Pro Met Thr Ser Pro Gly Ser Gly Leu Ile Leu

35 40 45

Gln Ala Asn Phe Val His Ser Gln Arg Arg Glu Ser Phe Leu Tyr Arg

50 55 60

Ser Asp Ser Asp Tyr Asp Leu Ser Pro Lys Ser Met Ser Arg Asn Ser

65 70 75 80

Ser Ile Ala Ser Asp Ile His Gly Asp Asp Leu Ile Val Thr Pro Phe

85 90 95

Ala Gln Val Leu Ala Ser Leu Arg Thr Val Arg Asn Asn Phe Ala Ala

100 105 110

Leu Thr Asn Leu Gln Asp Arg Ala Pro Ser Lys Arg Ser Pro Met Cys

115 120 125

Asn Gln Pro Ser Ile Asn Lys Ala Thr Ile Thr Glu Glu Ala Tyr Gln

130 135 140

Lys Leu Ala Ser Glu Thr Leu Glu Glu Leu Asp Trp Cys Leu Asp Gln

145 150 155 160

Leu Glu Thr Leu Gln Thr Arg His Ser Val Ser Glu Met Ala Ser Asn

165 170 175

Lys Phe Lys Arg Met Leu Asn Arg Glu Leu Thr His Leu Ser Glu Met

180 185 190

Ser Arg Ser Gly Asn Gln Val Ser Glu Phe Ile Ser Asn Thr Phe Leu

195 200 205

Asp Lys Gln His Glu Val Glu Ile Pro Ser Pro Thr Gln Lys Glu Lys

210 215 220

Glu Lys Lys Lys Arg Pro Met Ser Gln Ile Ser Gly Val Lys Lys Leu

225 230 235 240

Met His Ser Ser Ser Leu Thr Asn Ser Ser Ile Pro Arg Phe Gly Val

245 250 255

Lys Thr Glu Gln Glu Asp Val Leu Ala Lys Glu Leu Glu Asp Val Asn

260 265 270

Lys Trp Gly Leu His Val Phe Arg Ile Ala Glu Leu Ser Gly Asn Arg

275 280 285

Pro Leu Thr Val Ile Met His Thr Ile Phe Gln Glu Arg Asp Leu Leu

290 295 300

Lys Thr Phe Lys Ile Pro Val Asp Thr Leu Ile Thr Tyr Leu Met Thr

305 310 315 320

Leu Glu Asp His Tyr His Ala Asp Val Ala Tyr His Asn Asn Ile His

325 330 335

Ala Ala Asp Val Val Gln Ser Thr His Val Leu Leu Ser Thr Pro Ala

340 345 350

Leu Glu Ala Val Phe Thr Asp Leu Glu Ile Leu Ala Ala Ile Phe Ala

355 360 365

Ser Ala Ile His Asp Val Asp His Pro Gly Val Ser Asn Gln Phe Leu

370 375 380

Ile Asn Thr Asn Ser Glu Leu Ala Leu Met Tyr Asn Asp Ser Ser Val

385 390 395 400

Leu Glu Asn His His Leu Ala Val Gly Phe Lys Leu Leu Gln Glu Glu

405 410 415

Asn Cys Asp Ile Phe Gln Asn Leu Thr Lys Lys Gln Arg Gln Ser Leu

420 425 430

Arg Lys Met Val Ile Asp Ile Val Leu Ala Thr Asp Met Ser Lys His

435 440 445

Met Asn Leu Leu Ala Asp Leu Lys Thr Met Val Glu Thr Lys Lys Val

450 455 460

Thr Ser Ser Gly Val Leu Leu Leu Asp Asn Tyr Ser Asp Arg Ile Gln

465 470 475 480

Val Leu Gln Asn Met Val His Cys Ala Asp Leu Ser Asn Pro Thr Lys

485 490 495

Pro Leu Gln Leu Tyr Arg Gln Trp Thr Asp Arg Ile Met Glu Glu Phe

500 505 510

Phe Arg Gln Gly Asp Arg Glu Arg Glu Arg Gly Met Glu Ile Ser Pro

515 520 525

Met Cys Asp Lys His Asn Ala Ser Val Glu Lys Ser Gln Val Gly Phe

530 535 540

Ile Asp Tyr Ile Val His Pro Leu Trp Glu Thr Trp Ala Asp Leu Val

545 550 555 560

His Pro Asp Ala Gln Asp Ile Leu Asp Thr Leu Glu Asp Asn Arg Glu

565 570 575

Trp Tyr Gln Ser Thr Ile Pro Gln Ser Pro Ser Pro Ala Pro Asp Asp

580 585 590

Pro Glu Glu Gly Arg Gln Gly Gln Thr Glu Lys Phe Gln Phe Glu Leu

595 600 605

Thr Leu Glu Glu Asp Gly Glu Ser Asp Thr Glu Lys Asp Ser Gly Ser

610 615 620

Gln Val Glu Glu Asp Thr Ser Cys Ser Asp Ser Lys Thr Leu Cys Thr

625 630 635 640

Gln Asp Ser Glu Ser Thr Glu Ile Pro Leu Asp Glu Gln Val Glu Glu

645 650 655

Glu Ala Val Gly Glu Glu Glu Glu Ser Gln Pro Glu Ala Cys Val Ile

660 665 670

Asp Asp Arg Ser Pro Asp Thr

675

<210> 31

<211> 26

<212> DNA

<213> Artificial sequence

<220>

<223> PDE4D9_forward primer

<400> 31

atgagcatta ttatgaagcc aagatc 26

<210> 32

<211> 22

<212> DNA

<213> Artificial sequence

<220>

<223> PDE4D9_reverse primer

<400> 32

gtgccattgt ccacatcaaa ac 22

<210> 33

<211> 27

<212> DNA

<213> Artificial sequence

<220>

<223> PDE4D9 probe

<400> 33

ctacaagttc cctaaggact gcagagg 27

<210> 34

<211> 1435

<212> DNA

<213> Homo sapiens

<400> 34

ggcggggcct gcttctcctc agcttcaggc ggctgcgacg agccctcagg cgaacctctc 60

ggctttcccg cgcggcgccg cctcttgctg cgcctccgcc tcctcctctg ctccgccacc 120

ggcttcctcc tcctgagcag tcagcccgcg cgccggccgg ctccgttatg gcgacccgca 180

gccctggcgt cgtgattagt gatgatgaac caggttatga ccttgattta ttttgcatac 240

ctaatcatta tgctgaggat ttggaaaggg tgtttattcc tcatggacta attatggaca 300

ggactgaacg tcttgctcga gatgtgatga aggagatggg aggccatcac attgtagccc 360

tctgtgtgct caaggggggc tataaattct ttgctgacct gctggattac atcaaagcac 420

tgaatagaaa tagtgataga tccattccta tgactgtaga ttttatcaga ctgaaagct 480

attgtaatga ccagtcaaca ggggacataa aagtaattgg tggagatgat ctctcaactt 540

taactggaaa gaatgtcttg attgtggaag atataattga cactggcaaa acaatgcaga 600

ctttgctttc cttggtcagg cagtataatc caaagatggt caaggtcgca agcttgctgg 660

tgaaaaggac cccacgaagt gttggatata agccagactt tgttggattt gaaattccag 720

acaagtttgt tgtaggatat gcccttgact ataatgaata cttcagggat ttgaatcatg 780

tttgtgtcat tagtgaaact ggaaaagcaa aatacaaagc ctaagatgag agttcaagtt 840

gagtttggaa acatctggag tcctattgac atcgccagta aaattatcaa tgttctagtt 900

ctgtggccat ctgcttagta gagctttttg catgtatctt ctaagaattt tatctgtttt 960

gtactttaga aatgtcagtt gctgcattcc taaactgttt atttgcacta tgagcctata 1020

gactatcagt tccctttggg cggattgttg tttaacttgt aaatgaaaaa attctcttaa 1080

accacagcac tattgagtga aacattgaac tcatatctgt aagaaataaa gagaagatat 1140

1200

atattgttaa ttataccacc gtgtgttaga aaagtaagaa gcagtcaatt ttcacatcaa 1260

agacagcatc taagaagttt tgttctgtcc tggaattatt ttagtagtgt ttcagtaatg 1320

ttgactgtat tttccaactt gttcaaatta ttaccagtga atctttgtca gcagttccct 1380

tttaaatgca aatcaataaa ttcccaaaaa tttaaaaaaa aaaaaaaaaa aaaaa 1435

<210> 35

<211> 218

<212> PROTEIN

<213> Homo sapiens

<400> 35

Met Ala Thr Arg Ser Pro Gly Val Val Ile Ser Asp Asp Glu Pro Gly

1 5 10 15

Tyr Asp Leu Asp Leu Phe Cys Ile Pro Asn His Tyr Ala Glu Asp Leu

20 25 30

Glu Arg Val Phe Ile Pro His Gly Leu Ile Met Asp Arg Thr Glu Arg

35 40 45

Leu Ala Arg Asp Val Met Lys Glu Met Gly Gly His His Ile Val Ala

50 55 60

Leu Cys Val Leu Lys Gly Gly Tyr Lys Phe Phe Ala Asp Leu Leu Asp

65 70 75 80

Tyr Ile Lys Ala Leu Asn Arg Asn Ser Asp Arg Ser Ile Pro Met Thr

85 90 95

Val Asp Phe Ile Arg Leu Lys Ser Tyr Cys Asn Asp Gln Ser Thr Gly

100 105 110

Asp Ile Lys Val Ile Gly Gly Asp Asp Leu Ser Thr Leu Thr Gly Lys

115 120 125

Asn Val Leu Ile Val Glu Asp Ile Ile Asp Thr Gly Lys Thr Met Gln

130 135 140

Thr Leu Leu Ser Leu Val Arg Gln Tyr Asn Pro Lys Met Val Lys Val

145 150 155 160

Ala Ser Leu Leu Val Lys Arg Thr Pro Arg Ser Val Gly Tyr Lys Pro

165 170 175

Asp Phe Val Gly Phe Glu Ile Pro Asp Lys Phe Val Val Gly Tyr Ala

180 185 190

Leu Asp Tyr Asn Glu Tyr Phe Arg Asp Leu Asn His Val Cys Val Ile

195 200 205

Ser Glu Thr Gly Lys Ala Lys Tyr Lys Ala

210 215

<210> 36

<211> 24

<212> DNA

<213> Artificial sequence

<220>

<223> HPRT1_forward primer

<400> 36

gaggatttgg aaagggtgtt tatt 24

<210> 37

<211> 21

<212> DNA

<213> Artificial sequence

<220>

<223> HPRT1_reverse primer

<400> 37

acagaggggct acaatgtgat g 21

<210> 38

<211> 26

<212> DNA

<213> Artificial sequence

<220>

<223> HPRT1 probe

<400> 38

acgtcttgct cgagatgtga tgaagg 26

<210> 39

<211> 1771

<212> DNA

<213> Homo sapiens

<400> 39

ggcggccagg cggggcgcgg agtgggcgcg cggggccggga ggaggggcca gcgaccgcgg 60

caccgcctgt gcccgcccgc ccctccgcag ccgctactta agaggctcca gcgccggccc 120

cgccctagtg cgttacttac ctcgactctt agcttgtcgg ggacggtaac cgggacccgg 180

tgtctgctcc tgtcgccttc gcctcctaat ccctagccac tatgcgtgag tgcatctcca 240

tccacgttgg ccaggctggt gtccagattg gcaatgcctg ctgggagctc tactgcctgg 300

360

actccttcaa caccttcttc agtgagacgg gcgctggcaa gcacgtgccc cgggctgtgt 420

ttgtagactt ggaacccaca gtcattgatg aagttcgcac tggcacctac cgccagctct 480

tccaccctga gcagctcatc acaggcaagg aagatgctgc caataactat gcccgagggc 540

actacaccat tggcaaggag atcattgacc ttgtgttgga ccgaattcgc aagctggctg 600

accagtgcac cggtcttcag ggcttcttgg ttttccacag ctttggtggg ggaactggtt 660

ctgggttcac ctccctgctc atggaacgtc tctcagttga ttatggcaag aagtccaagc 720

tggagttctc catttaccca gcaccccagg tttccacagc tgtagttgag ccctacaact 780

ccatcctcac cacccacacc accctggagc actctgattg tgccttcatg gtagacaatg 840

aggccatcta tgacatctgt cgtagaaacc tcgatatcga gcgcccaacc tacactaacc 900

ttaaccgcct tattagccag attgtgtcct ccatcactgc ttccctgaga tttgatggag 960

ccctgaatgt tgacctgaca gaattccaga ccaacctggt gccctacccc cgcatccact 1020

tccctctggc cacatatgcc cctgtcatct ctgctgagaa agcctaccat gaacagcttt 1080

ctgtagcaga gatcaccaat gcttgctttg agccagccaa ccagatggtg aaatgtgacc 1140

ctcgccatgg taaatacatg gcttgctgcc tgttgtaccg tggtgacgtg gttcccaaag 1200

atgtcaatgc tgccattgcc accatcaaaa ccaagcgcag catccagttt gtggattggt 1260

gccccactgg cttcaaggtt ggcatcaact accagcctcc cactgtggtg cctggtggag 1320

acctggccaa ggtacagaga gctgtgtgca tgctgagcaa caccacagcc attgctgagg 1380

cctgggctcg cctggaccac aagtttgacc tgatgtatgc caagcgtgcc tttgttcact 1440

ggtacgtggg tgaggggatg gaggaaggcg agttttcaga ggcccgtgaa gatatggctg 1500

1560

aggagagga atactaatta tccattcctt ttggccctgc agcatgtcat gctcccagaa 1620

tttcagcttc agcttaactg acagacgtta aagctttctg gttagattgt tttcacttgg 1680

tgatcatgtc ttttccatgt gtacctgtaa tatttttcca tcatatctca aagtaaagtc 1740

attaacatca aaaaaaaaaa aaaaaaaaaa a 1771

<210> 40

<211> 451

<212> PROTEIN

<213> Homo sapiens

<400> 40

Met Arg Glu Cys Ile Ser Ile His Val Gly Gln Ala Gly Val Gln Ile

1 5 10 15

Gly Asn Ala Cys Trp Glu Leu Tyr Cys Leu Glu His Gly Ile Gln Pro

20 25 30

Asp Gly Gln Met Pro Ser Asp Lys Thr Ile Gly Gly Gly Asp Asp Ser

35 40 45

Phe Asn Thr Phe Phe Ser Glu Thr Gly Ala Gly Lys His Val Pro Arg

50 55 60

Ala Val Phe Val Asp Leu Glu Pro Thr Val Ile Asp Glu Val Arg Thr

65 70 75 80

Gly Thr Tyr Arg Gln Leu Phe His Pro Glu Gln Leu Ile Thr Gly Lys

85 90 95

Glu Asp Ala Ala Asn Asn Tyr Ala Arg Gly His Tyr Thr Ile Gly Lys

100 105 110

Glu Ile Ile Asp Leu Val Leu Asp Arg Ile Arg Lys Leu Ala Asp Gln

115 120 125

Cys Thr Gly Leu Gln Gly Phe Leu Val Phe His Ser Phe Gly Gly Gly

130 135 140

Thr Gly Ser Gly Phe Thr Ser Leu Leu Met Glu Arg Leu Ser Val Asp

145 150 155 160

Tyr Gly Lys Lys Ser Lys Leu Glu Phe Ser Ile Tyr Pro Ala Pro Gln

165 170 175

Val Ser Thr Ala Val Val Glu Pro Tyr Asn Ser Ile Leu Thr His

180 185 190

Thr Thr Leu Glu His Ser Asp Cys Ala Phe Met Val Asp Asn Glu Ala

195 200 205

Ile Tyr Asp Ile Cys Arg Arg Asn Leu Asp Ile Glu Arg Pro Thr Tyr

210 215 220

Thr Asn Leu Asn Arg Leu Ile Ser Gln Ile Val Ser Ser Ile Thr Ala

225 230 235 240

Ser Leu Arg Phe Asp Gly Ala Leu Asn Val Asp Leu Thr Glu Phe Gln

245 250 255

Thr Asn Leu Val Pro Tyr Pro Arg Ile His Phe Pro Leu Ala Thr Tyr

260 265 270

Ala Pro Val Ile Ser Ala Glu Lys Ala Tyr His Glu Gln Leu Ser Val

275 280 285

Ala Glu Ile Thr Asn Ala Cys Phe Glu Pro Ala Asn Gln Met Val Lys

290 295 300

Cys Asp Pro Arg His Gly Lys Tyr Met Ala Cys Cys Leu Leu Tyr Arg

305 310 315 320

Gly Asp Val Val Pro Lys Asp Val Asn Ala Ala Ile Ala Thr Ile Lys

325 330 335

Thr Lys Arg Ser Ile Gln Phe Val Asp Trp Cys Pro Thr Gly Phe Lys

340 345 350

Val Gly Ile Asn Tyr Gln Pro Pro Thr Val Val Pro Gly Gly Asp Leu

355 360 365

Ala Lys Val Gln Arg Ala Val Cys Met Leu Ser Asn Thr Thr Ala Ile

370 375 380

Ala Glu Ala Trp Ala Arg Leu Asp His Lys Phe Asp Leu Met Tyr Ala

385 390 395 400

Lys Arg Ala Phe Val His Trp Tyr Val Gly Glu Gly Met Glu Glu Gly

405 410 415

Glu Phe Ser Glu Ala Arg Glu Asp Met Ala Ala Leu Glu Lys Asp Tyr

420 425 430

Glu Glu Val Gly Val Asp Ser Val Glu Gly Glu Gly Glu Glu Glu Glu Gly

435 440 445

Glu Glu Tyr

450

<210> 41

<211> 22

<212> DNA

<213> Artificial sequence

<220>

<223> TUBA1B_forward primer

<400> 41

tgactccttc aacaccttct tc 22

<210> 42

<211> 18

<212> DNA

<213> Artificial sequence

<220>

<223> TUBA1B_reverse primer

<400> 42

tgccagtgcg aacttcat 18

<210> 43

<211> 24

<212> DNA

<213> Artificial sequence

<220>

<223> TUBA1B probe

<400> 43

ccgggctgtg tttgtagact tgga 24

<210> 44

<211> 5416

<212> DNA

<213> Homo sapiens

<400> 44

agtgggccgc catgttgtcg gagtgaaagg taagggggag cgagagcgcc agagagagaa 60

gatcgggggg ctgaaatcca tcttcatcct accgctccgc ccgtgttggg ggaatgagcg 120

ttgcatgtgt cttgaagaga aaagcagtgc tttggcagga ctctttcagc ccccacctga 180

aacatcaccc tcaagaacca gctaatccca acatgcctgt tgttttgaca tctggaacag 240

ggtcgcaagc gcagccacaa ccagctgcaa atcaggctct tgcagctggg actcactcca 300

gccctgtccc aggatctata ggagttgcag gccgttccca ggacgacgct atggtggact 360

acttctttca gaggcagcat ggtgagcagc ttgggggagg aggaagtgga ggaggcggct 420

ataataatag caaacatcga tggcctactg gggataacat tcatgcagaa catcaggtgc 480

gttccatgga tgaactgaat catgattttc aagcacttgc tctggaggga agagcgatgg 540

gagagcagct cttgccaggt aaaaagtttt gggaaacaga tgaatccagc aaagatggac 600

caaaaggaat attcctgggt gatcaatggc gagacagtgc ctggggaaca tcagatcatt 660

cagtttccca gccaatcatg gtgcagagaa gacctggtca gagtttccat gtgaacagtg 720

aggtcaattc tgtactgtcc ccacgatcgg agagtggggg actaggcgtt agcatggtgg 780

agtatgtgtt gagctcatcc ccgggcgatt cctgtctaag aaaagggagga tttggcccaa 840

gggatgcaga cagtgatgaa aacgacaaag gtgaaaagaa gaacaagggt acgtttgatg 900

gagataagct aggagatttg aaggaggagg gtgatgtgat ggacaagacc aatggtttac 960

cagtgcagaa tgggattgat gcagacgtca aagattttag ccgtacccct ggtaattgcc 1020

agaactctgc taatgaagtg gatcttctgg gtccaaacca gaatggttct gagggcttag 1080

cccagctgac cagcaccaat ggtgccaagc ctgtggagga tttctccaac atggagtccc 1140

agagtgtccc cttggacccc atggaacatg tgggcatgga gcctcttcag tttgattatt 1200

caggcacgca ggtacctgtg gactcagcag cagcaactgt gggacttttt gactacaatt 1260

ctcaacaaca gctgttccaa agacctaatg cgcttgctgt ccagcagttg acagctgctc 1320

agcagcagca gtatgcactg gcagctgctc atcagccgca catcggttta gctcccgctg 1380

cgtttgtccc caatccatac atcatcagcg ctgctccccc agggacggac ccctacacag 1440

ctggattggc tgcagcagcg acactaggcc cagctgtggt ccctcaccag tattatggag 1500

ttactccctg gggagtctac cctgccagtc ttttccagca gcaagctgcc gctgccgctg 1560

cagcaactaa ttcagctaat caacagacca ccccacaggc tcagcaagga cagcagcagg 1620

ttctccgtgg aggagccagc caacgtcctt tgaccccaaa ccagaaccag cagggacagc 1680

aaacggatcc ccttgtggca gctgcagcag tgaattctgc ccttgcattt ggacaaggtc 1740

tggcagcagg catgccaggt tatccggtgt tggctcctgc tgcttactat gaccaaactg 1800

gtgcccttgt agtgaatgca ggcgcgagaa atggtcttgg agctcctgtt cgacttgtag 1860

ctcctgcccc agtcatcatt agttcctcag ctgcacaagc agctgttgca gcagccgcag 1920

cttcagcaaa tggagcagct ggtggtcttg ctggaacaac aaatggacca tttcgccctt 1980

taggaacaca gcagcctcag ccccagcccc agcagcagcc caataacaac ctggcatcca 2040

gttctttcta cggcaacaac tctctgaaca gcaattcaca gagcagctcc ctcttctccc 2100

agggctctgc ccagcctgcc aacacatcct tgggattcgg aagtagcagt tctctcggcg 2160

ccaccctggg atccgccctt ggagggtttg gaacagcagt tgcaaactcc aacactggca 2220

gtggctcccg ccgtgactcc ctgactggca gcagtgacct ttataagagg acatcgagca 2280

gcttgacccc cattggacac agtttttata acggccttag ctttttcctcc tctcctggac 2340

ccgtgggcat gcctctccct agtcagggac caggacattc acagacacca cctccttccc 2400

tctcttcaca tggatcctct tcaagcttaa acctgggagg actcacgaat ggcagtggaa 2460

gatacatctc tgctgctcca ggcgctgaag ccaagtaccg cagtgcaagc agcgcctcca 2520

gcctcttcag cccgagcagc actcttttct cttcctctcg tttgcgatat ggaatgtctg 2580

atgtcatgcc ttctggcagg agcaggcttt tggaagattt tcgaaacaac cggtacccca 2640

attacaact gcgggagatt gctggacata taatggaatt ttcccaagac cagcatgggt 2700

ccagattcat tcagctgaaa ctggagcgtg ccacaccagc tgagcgccag cttgtcttca 2760

atgaaatcct ccaggctgcc taccaactca tggtggatgt gtttggtaat tacgtcattc 2820

agaagttctt tgaatttggc agtcttgaac agaagctggc tttggcagaa cggattcgag 2880

gccacgtcct gtcattggca ctacagatgt atggctgccg tgttatccag aaagctcttg 2940

agtttattcc ttcagaccag caggtaatta atgagatggt tcgggaacta gatggccatg 3000

3060

tacagcccca gtctttgcaa tttatcatcg atgcgtttaa gggacaggta tttgccttat 3120

ccacacatcc ttatggctgc cgagtgattc agagaatcct ggagcactgt ctccctgacc 3180

agacactccc tattttagag gagcttcacc agcacacaga gcagcttgta caggatcaat 3240

atggaaatta tgtaatccaa catgtactgg agcacggtcg tcctgaggat aaaagcaaaa 3300

ttgtagcaga aatccgaggc aatgtacttg tattgagtca gcacaaattt gcaagcaatg 3360

ttgtggagaa gtgtgttact cacgcctcac gtacggagcg cgctgtgctc atcgatgagg 3420

3480

ccaactacgt ggtccagaag atgattgacg tggcggagcc aggccagcgg aagatcgtca 3540

tgcataagat ccggccccac atcgcaactc ttcgtaagta cacctatggc aagcacattc 3600

tggccaagct ggagaagtac tacatgaaga acggtgttga cttagggccc atctgtggcc 3660

cccctaatgg tatcatctga ggcagtgtca cccgctgttc cctcattccc gctgacctca 3720

ctggcccact ggcaaatcca accagcaacc agaaatgttc tagtgtagag tctgagacgg 3780

gcaagtggtt gctccaggat tactccctcc tccaaaaaag gaatcaaatc cacgagtgga 3840

aaagcctttg taaatttaat ttttattacac ataacatgta ctattttttt taattgacta 3900

attgccctgc tgttttactg gtgtatagga tacttgtaca taggtaacca atgtacatgg 3960

4020

gttggtttaa caaaaaaaaa aagctttaaa aaaaaaagaa aaaaaggaaa aggtttttag 4080

ctcatttgcc tggccggcaa gttttgcaaa tagctcttcc ccacctcctc attttagtaa 4140

aaaacaaaca aaaacaaaaa aacctgagaa gtttgaattg tagttaaatg accccaaact 4200

ggcatttaac actgtttata aaaaatatat atatatatat atatatatat aatgaaaaag 4260

4320

cttttttgga gactatatta tgctgaagaa ggctgttcgt gaggaggaga tgcgagcacc 4380

4440

ttaacattga aaggtaaaat ctgattattt agcatgagaa aaaaaaatcc aactctgctt 4500

ttggtcttgc ttctataaat atatagtgta tacttggtgt agactttgca tatatacaaa 4560

tttgtagtat tttcttgttt tgatgtctaa tctgtatcta taatgtaccc tagtagtcga 4620

acatactttt gattgtacaa ttgtacattt gtatacctgt aatgtaaatg tggagaagtt 4680

tgaatcaaca taaacacgtt ttttggtaag aaaagagaat tagccagccc tgtgcattca 4740

gtgtatattc tcacctttta tggtcgtagc atatagtgtt gtatattgta aattgtaatt 4800

tcaaccagaa gtaaattttt ttcttttgaa ggaataaatg ttctttatac agcctagtta 4860

atgtttaaaa agaaaaaaat agcttggttt tatttgtcat ctagtctcaa gtatagcgag 4920

attctttcta aatgttattc aagattgagt tctcactagt gtttttttaa tcctaaaaaa 4980

gtaatgtttt gattttgtga cagtcaaaag gacgtgcaaa agtctagcct tgcccgagct 5040

ttccttacaa tcagagcccc tctcaccttg taaagtgtga atcgcccttc ccttttgtac 5100

agaagatgaa ctgtattttg cattttgtct acttgtaagt gaatgtaaca tactgtcaat 5160

tttccttgtt tgaatataga attgtaacac tacacggtgt acatttccag agccttgtgt 5220

atatttccaa tgaacttttt tgcaagcaca cttgtaacca tatgtgtata attaacaaac 5280

ctgtgtatgc ttatgcctgg gcaactattt tttgtaactc ttgtgtagat tgtctctaaa 5340

caatgtgtga tctttatttt gaaaaataca gaactttgga atctgaaaaa aaaaaaaaaa 5400

aaaaaaaaaaaaaaaa 5416

<210> 45

<211> 5410

<212> DNA

<213> Homo sapiens

<400> 45

agtgggccgc catgttgtcg gagtgaaagg taagggggag cgagagcgcc agagagagaa 60

gatcgggggg ctgaaatcca tcttcatcct accgctccgc ccgtgttggg ggaatgagcg 120

ttgcatgtgt cttgaagaga aaagcagtgc tttggcagga ctctttcagc ccccacctga 180

aacatcaccc tcaagaacca gctaatccca acatgcctgt tgttttgaca tctggaacag 240

ggtcgcaagc gcagccacaa ccagctgcaa atcaggctct tgcagctggg actcactcca 300

gccctgtccc aggatctata ggagttgcag gccgttccca ggacgacgct atggtggact 360

acttctttca gaggcagcat ggtgagcagc ttgggggagg aggaagtgga ggaggcggct 420

ataataatag caaacatcga tggcctactg gggataacat tcatgcagaa catcaggtgc 480

gttccatgga tgaactgaat catgattttc aagcacttgc tctggaggga agagcgatgg 540

gagagcagct cttgccaggt aaaaagtttt gggaaacaga tgaatccagc aaagatggac 600

caaaaggaat attcctgggt gatcaatggc gagacagtgc ctggggaaca tcagatcatt 660

cagtttccca gccaatcatg gtgcagagaa gacctggtca gagtttccat gtgaacagtg 720

aggtcaattc tgtactgtcc ccacgatcgg agagtggggg actaggcgtt agcatggtgg 780

agtatgtgtt gagctcatcc ccgggcgatt cctgtctaag aaaagggagga tttggcccaa 840

gggatgcaga cagtgatgaa aacgacaaag gtgaaaagaa gaacaagggt acgtttgatg 900

gagataagct aggagatttg aaggaggagg gtgatgtgat ggacaagacc aatggtttac 960

cagtgcagaa tgggattgat gcagacgtca aagattttag ccgtacccct ggtaattgcc 1020

agaactctgc taatgaagtg gatcttctgg gtccaaacca gaatggttct gagggcttag 1080

cccagctgac cagcaccaat ggtgccaagc ctgtggagga tttctccaac atggagtccc 1140

agagtgtccc cttggacccc atggaacatg tgggcatgga gcctcttcag tttgattatt 1200

caggcacgca ggtacctgtg gactcagcag cagcaactgt gggacttttt gactacaatt 1260

ctcaacaaca gctgttccaa agacctaatg cgcttgctgt ccagcagttg acagctgctc 1320

agcagcagca gtatgcactg gcagctgctc atcagccgca catcggttta gctcccgctg 1380

cgtttgtccc caatccatac atcatcagcg ctgctccccc agggacggac ccctacacag 1440

ctggattggc tgcagcagcg acactaggcc cagctgtggt ccctcaccag tattatggag 1500

ttactccctg gggagtctac cctgccagtc ttttccagca gcaagctgcc gctgccgctg 1560

cagcaactaa ttcagctaat caacagacca ccccacaggc tcagcaagga cagcagcagg 1620

ttctccgtgg aggagccagc caacgtcctt tgaccccaaa ccagaaccag cagggacagc 1680

aaacggatcc ccttgtggca gctgcagcag tgaattctgc ccttgcattt ggacaaggtc 1740

tggcagcagg catgccaggt tatccggtgt tggctcctgc tgcttactat gaccaaactg 1800

gtgcccttgt agtgaatgca ggcgcgagaa atggtcttgg agctcctgtt cgacttgtag 1860

ctcctgcccc agtcatcatt agttcctcag ctgcacaagc agctgttgca gcagccgcag 1920

cttcagcaaa tggagcagct ggtggtcttg ctggaacaac aaatggacca tttcgccctt 1980

taggaacaca gcagcctcag ccccagcccc agcagcagcc caataacaac ctggcatcca 2040

gttctttcta cggcaacaac tctctgaaca gcaattcaca gagcagctcc ctcttctccc 2100

agggctctgc ccagcctgcc aacacatcct tgggattcgg aagtagcagt tctctcggcg 2160

ccaccctggg atccgccctt ggagggtttg gaacagcagt tgcaaactcc aacactggca 2220

gtggctcccg ccgtgactcc ctgactggca gcagtgacct ttataagagg acatcgagca 2280

gcttgacccc cattggacac agtttttata acggccttag ctttttcctcc tctcctggac 2340

ccgtgggcat gcctctccct agtcagggac caggacattc acagacacca cctccttccc 2400

tctcttcaca tggatcctct tcaagcttaa acctgggagg actcacgaat ggcagtggaa 2460

gatacatctc tgctgctcca ggcgctgaag ccaagtaccg cagtgcaagc agcgcctcca 2520

gcctcttcag cccgagcagc actcttttct cttcctctcg tttgcgatat ggaatgtctg 2580

atgtcatgcc ttctggcagg agcaggcttt tggaagattt tcgaaacaac cggtacccca 2640

attacaact gcgggagatt gctggacata taatggaatt ttcccaagac cagcatgggt 2700

ccagattcat tcagctgaaa ctggagcgtg ccacaccagc tgagcgccag cttgtcttca 2760

atgaaatcct ccaggctgcc taccaactca tggtggatgt gtttggtaat tacgtcattc 2820

agaagttctt tgaatttggc agtcttgaac agaagctggc tttggcagaa cggattcgag 2880

gccacgtcct gtcattggca ctacagatgt atggctgccg tgttatccag aaagctcttg 2940

agtttattcc ttcagaccag cagaatgaga tggttcggga actagatggc catgtcttga 3000

3060

3120

atccttatgg ctgccgagtg attcagagaa tcctggagca ctgtctccct gaccagacac 3180

3240

attatgtaat ccaacatgta ctggagcacg gtcgtcctga ggataaaagc aaaattgtag 3300

cagaaatccg aggcaatgta cttgtattga gtcagcacaa atttgcaagc aatgttgtgg 3360

agaagtgtgt tactcacgcc tcacgtacgg agcgcgctgt gctcatcgat gaggtgtgca 3420

ccatgaacga cggtccccac agtgccttat aaccatgat gaaggaccag tatgccaact 3480

acgtggtcca gaagatgatt gacgtggcgg agccaggcca gcggaagatc gtcatgcata 3540

agatccggcc ccacatcgca actcttcgta agtacaccta tggcaagcac attctggcca 3600

agctggagaa gtactacatg aagaacggtg ttgacttagg gcccatctgt ggccccccta 3660

atggtatcat ctgaggcagt gtcacccgct gttccctcat tcccgctgac ctcactggcc 3720

3780 cactggcaaa tccaaccagc aaccagaaat gttctagtgt

ggttgctcca ggattactcc ctcctccaaa aaaggaatca aatccacgag tggaaaagcc 3840

tttgtaaatt taattttatt acacataaca tgtactattt tttttaattg actaattgcc 3900

ctgctgtttt actggtgtat aggatacttg tacataggta accaatgtac atgggaggcc 3960

acatattttg ttcactgttg tatctatatt tcacatgtgg aaactttcag ggtggttggt 4020

ttaacaaaaa aaaaaagctt taaaaaaaaa agaaaaaaag gaaaaggtttt ttagctcatt 4080

tgcctggccg gcaagttttg caaaatagctc ttccccacct cctcatttta gtaaaaaaca 4140

aacaaaaaca aaaaaacctg agaagtttga attgtagtta aatgacccca aactggcatt 4200

taacactgtt tataaaaaat atatatatat atatatatat atataatgaa aaaggtttca 4260

4320

tggagactat attatgctga agaaggctgt tcgtgaggag gagatgcgag cacccagaac 4380

gtcttttgag gctgggcggg tgtgattgtt tactgcctac tggattttt tctattaaca 4440

ttgaaaggta aaatctgatt atttagcatg agaaaaaaaa atccaactct gcttttggtc 4500

ttgcttctat aaattatag tgtatacttg gtgtagactt tgcatatata caaatttgta 4560

gtattttctt gttttgatgt ctaatctgta tctataatgt accctagtag tcgaacatac 4620

ttttgattgt acaattgtac atttgtatac ctgtaatgta aatgtggaga agtttgaatc 4680

aacataaaca cgttttttgg taagaaaaga gaattagcca gccctgtgca ttcagtgtat 4740

attctcacct tttatggtcg tagcatatag tgttgtatat tgtaaattgt aatttcaacc 4800

agaagtaaat ttttttcttt tgaaggaata aatgttcttt atacagccta gttaatgttt 4860

aaaaagaaaa aaatagcttg gttttatttg tcatctagtc tcaagtatag cgagattctt 4920

tctaaatgtt attcaagatt gagttctcac tagtgttttt ttaatcctaa aaaagtaatg 4980

ttttgatttt gtgacagtca aaaggacgtg caaaagtcta gccttgcccg agctttcctt 5040

acaatcagag cccctctcac cttgtaaagt gtgaatcgcc cttccctttt gtacagaaga 5100

tgaactgtat tttgcatttt gtctacttgt aagtgaatgt aacatactgt caattttcct 5160

tgtttgaata tagaattgta acactacacg gtgtacattt cgagcctt gtgtatattt 5220

ccaatgaact ttttgcaag cacacttgta accatatgtg tataattaac aaacctgtgt 5280

atgcttatgc ctgggcaact attttttgta actcttgtgt agattgtctc taaacaatgt 5340

gtgatcttta ttttgaaaaa tacagaactt tggaatctga aaaaaaaaaa aaaaaaaaaa 5400

aaaaaaaaaa 5410

<210> 46

<211> 1188

<212> PROTEIN

<213> Homo sapiens

<400> 46

Met Ser Val Ala Cys Val Leu Lys Arg Lys Ala Val Leu Trp Gln Asp

1 5 10 15

Ser Phe Ser Pro His Leu Lys His His Pro Gln Glu Pro Ala Asn Pro

20 25 30

Asn Met Pro Val Val Leu Thr Ser Gly Thr Gly Ser Gln Ala Gln Pro

35 40 45

Gln Pro Ala Ala Asn Gln Ala Leu Ala Ala Gly Thr His Ser Ser Pro

50 55 60

Val Pro Gly Ser Ile Gly Val Ala Gly Arg Ser Gln Asp Asp Ala Met

65 70 75 80

Val Asp Tyr Phe Phe Gln Arg Gln His Gly Glu Gln Leu Gly Gly Gly

85 90 95

Gly Ser Gly Gly Gly Gly Tyr Asn Asn Ser Lys His Arg Trp Pro Thr

100 105 110

Gly Asp Asn Ile His Ala Glu His Gln Val Arg Ser Met Asp Glu Leu

115 120 125

Asn His Asp Phe Gln Ala Leu Ala Leu Glu Gly Arg Ala Met Gly Glu

130 135 140

Gln Leu Leu Pro Gly Lys Lys Phe Trp Glu Thr Asp Glu Ser Ser Lys

145 150 155 160

Asp Gly Pro Lys Gly Ile Phe Leu Gly Asp Gln Trp Arg Asp Ser Ala

165 170 175

Trp Gly Thr Ser Asp His Ser Val Ser Gln Pro Ile Met Val Gln Arg

180 185 190

Arg Pro Gly Gln Ser Phe His Val Asn Ser Glu Val Asn Ser Val Leu

195 200 205

Ser Pro Arg Ser Glu Ser Gly Gly Leu Gly Val Ser Met Val Glu Tyr

210 215 220

Val Leu Ser Ser Ser Pro Gly Asp Ser Cys Leu Arg Lys Gly Gly Phe

225 230 235 240

Gly Pro Arg Asp Ala Asp Ser Asp Glu Asn Asp Lys Gly Glu Lys Lys

245 250 255

Asn Lys Gly Thr Phe Asp Gly Asp Lys Leu Gly Asp Leu Lys Glu Glu

260 265 270

Gly Asp Val Met Asp Lys Thr Asn Gly Leu Pro Val Gln Asn Gly Ile

275 280 285

Asp Ala Asp Val Lys Asp Phe Ser Arg Thr Pro Gly Asn Cys Gln Asn

290 295 300

Ser Ala Asn Glu Val Asp Leu Leu Gly Pro Asn Gln Asn Gly Ser Glu

305 310 315 320

Gly Leu Ala Gln Leu Thr Ser Thr Asn Gly Ala Lys Pro Val Glu Asp

325 330 335

Phe Ser Asn Met Glu Ser Gln Ser Val Pro Leu Asp Pro Met Glu His

340 345 350

Val Gly Met Glu Pro Leu Gln Phe Asp Tyr Ser Gly Thr Gln Val Pro

355 360 365

Val Asp Ser Ala Ala Ala Thr Val Gly Leu Phe Asp Tyr Asn Ser Gln

370 375 380

Gln Gln Leu Phe Gln Arg Pro Asn Ala Leu Ala Val Gln Gln Leu Thr

385 390 395 400

Ala Ala Gln Gln Gln Gln Tyr Ala Leu Ala Ala Ala His Gln Pro His

405 410 415

Ile Gly Leu Ala Pro Ala Ala Phe Val Pro Asn Pro Tyr Ile Ile Ser

420 425 430

Ala Ala Pro Pro Gly Thr Asp Pro Tyr Thr Ala Gly Leu Ala Ala Ala

435 440 445

Ala Thr Leu Gly Pro Ala Val Val Pro His Gln Tyr Tyr Gly Val Thr

450 455 460

Pro Trp Gly Val Tyr Pro Ala Ser Leu Phe Gln Gln Gln Ala Ala Ala

465 470 475 480

Ala Ala Ala Ala Thr Asn Ser Ala Asn Gln Gln Thr Thr Pro Gln Ala

485 490 495

Gln Gln Gly Gln Gln Gln Val Leu Arg Gly Gly Ala Ser Gln Arg Pro

500 505 510

Leu Thr Pro Asn Gln Asn Gln Gln Gly Gln Gln Thr Asp Pro Leu Val

515 520 525

Ala Ala Ala Ala Val Asn Ser Ala Leu Ala Phe Gly Gln Gly Leu Ala

530 535 540

Ala Gly Met Pro Gly Tyr Pro Val Leu Ala Pro Ala Ala Tyr Tyr Asp

545 550 555 560

Gln Thr Gly Ala Leu Val Val Asn Ala Gly Ala Arg Asn Gly Leu Gly

565 570 575

Ala Pro Val Arg Leu Val Ala Pro Ala Pro Val Ile Ile Ser Ser Ser

580 585 590

Ala Ala Gln Ala Ala Val Ala Ala Ala Ala Ala Ser Ala Asn Gly Ala

595 600 605

Ala Gly Gly Leu Ala Gly Thr Thr Asn Gly Pro Phe Arg Pro Leu Gly

610 615 620

Thr Gln Gln Pro Gln Pro Gln Pro Gln Gln Gln Pro Asn Asn Asn Leu

625 630 635 640

Ala Ser Ser Ser Phe Tyr Gly Asn Asn Ser Leu Asn Ser Asn Ser Gln

645 650 655

Ser Ser Ser Leu Phe Ser Gln Gly Ser Ala Gln Pro Ala Asn Thr Ser

660 665 670

Leu Gly Phe Gly Ser Ser Ser Ser Leu Gly Ala Thr Leu Gly Ser Ala

675 680 685

Leu Gly Gly Phe Gly Thr Ala Val Ala Asn Ser Asn Thr Gly Ser Gly

690 695 700

Ser Arg Arg Asp Ser Leu Thr Gly Ser Ser Asp Leu Tyr Lys Arg Thr

705 710 715 720

Ser Ser Ser Leu Thr Pro Ile Gly His Ser Phe Tyr Asn Gly Leu Ser

725 730 735

Phe Ser Ser Ser Pro Gly Pro Val Gly Met Pro Leu Pro Ser Gln Gly

740 745 750

Pro Gly His Ser Gln Thr Pro Pro Pro Ser Leu Ser Ser His Gly Ser

755 760 765

Ser Ser Ser Leu Asn Leu Gly Gly Leu Thr Asn Gly Ser Gly Arg Tyr

770 775 780

Ile Ser Ala Ala Pro Gly Ala Glu Ala Lys Tyr Arg Ser Ala Ser Ser

785 790 795 800

Ala Ser Ser Leu Phe Ser Pro Ser Ser Thr Leu Phe Ser Ser Ser Arg

805 810 815

Leu Arg Tyr Gly Met Ser Asp Val Met Pro Ser Gly Arg Ser Arg Leu

820 825 830

Leu Glu Asp Phe Arg Asn Asn Arg Tyr Pro Asn Leu Gln Leu Arg Glu

835 840 845

Ile Ala Gly His Ile Met Glu Phe Ser Gln Asp Gln His Gly Ser Arg

850 855 860

Phe Ile Gln Leu Lys Leu Glu Arg Ala Thr Pro Ala Glu Arg Gln Leu

865 870 875 880

Val Phe Asn Glu Ile Leu Gln Ala Ala Tyr Gln Leu Met Val Asp Val

885 890 895

Phe Gly Asn Tyr Val Ile Gln Lys Phe Phe Glu Phe Gly Ser Leu Glu

900 905 910

Gln Lys Leu Ala Leu Ala Glu Arg Ile Arg Gly His Val Leu Ser Leu

915 920 925

Ala Leu Gln Met Tyr Gly Cys Arg Val Ile Gln Lys Ala Leu Glu Phe

930 935 940

Ile Pro Ser Asp Gln Gln Val Ile Asn Glu Met Val Arg Glu Leu Asp

945 950 955 960

Gly His Val Leu Lys Cys Val Lys Asp Gln Asn Gly Asn His Val Val

965 970 975

Gln Lys Cys Ile Glu Cys Val Gln Pro Gln Ser Leu Gln Phe Ile Ile

980 985 990

Asp Ala Phe Lys Gly Gln Val Phe Ala Leu Ser Thr His Pro Tyr Gly

995 1000 1005

Cys Arg Val Ile Gln Arg Ile Leu Glu His Cys Leu Pro Asp Gln

1010 1015 1020

Thr Leu Pro Ile Leu Glu Glu Leu His Gln His Thr Glu Gln Leu

1025 1030 1035

Val Gln Asp Gln Tyr Gly Asn Tyr Val Ile Gln His Val Leu Glu

1040 1045 1050

His Gly Arg Pro Glu Asp Lys Ser Lys Ile Val Ala Glu Ile Arg

1055 1060 1065

Gly Asn Val Leu Val Leu Ser Gln His Lys Phe Ala Ser Asn Val

1070 1075 1080

Val Glu Lys Cys Val Thr His Ala Ser Arg Thr Glu Arg Ala Val

1085 1090 1095

Leu Ile Asp Glu Val Cys Thr Met Asn Asp Gly Pro His Ser Ala

1100 1105 1110

Leu Tyr Thr Met Met Lys Asp Gln Tyr Ala Asn Tyr Val Val Gln

1115 1120 1125

Lys Met Ile Asp Val Ala Glu Pro Gly Gln Arg Lys Ile Val Met

1130 1135 1140

His Lys Ile Arg Pro His Ile Ala Thr Leu Arg Lys Tyr Thr Tyr

1145 1150 1155

Gly Lys His Ile Leu Ala Lys Leu Glu Lys Tyr Tyr Met Lys Asn

1160 1165 1170

Gly Val Asp Leu Gly Pro Ile Cys Gly Pro Pro Asn Gly Ile Ile

1175 1180 1185

<210> 47

<211> 1186

<212> PROTEIN

<213> Homo sapiens

<400> 47

Met Ser Val Ala Cys Val Leu Lys Arg Lys Ala Val Leu Trp Gln Asp

1 5 10 15

Ser Phe Ser Pro His Leu Lys His His Pro Gln Glu Pro Ala Asn Pro

20 25 30

Asn Met Pro Val Val Leu Thr Ser Gly Thr Gly Ser Gln Ala Gln Pro

35 40 45

Gln Pro Ala Ala Asn Gln Ala Leu Ala Ala Gly Thr His Ser Ser Pro

50 55 60

Val Pro Gly Ser Ile Gly Val Ala Gly Arg Ser Gln Asp Asp Ala Met

65 70 75 80

Val Asp Tyr Phe Phe Gln Arg Gln His Gly Glu Gln Leu Gly Gly Gly

85 90 95

Gly Ser Gly Gly Gly Gly Tyr Asn Asn Ser Lys His Arg Trp Pro Thr

100 105 110

Gly Asp Asn Ile His Ala Glu His Gln Val Arg Ser Met Asp Glu Leu

115 120 125

Asn His Asp Phe Gln Ala Leu Ala Leu Glu Gly Arg Ala Met Gly Glu

130 135 140

Gln Leu Leu Pro Gly Lys Lys Phe Trp Glu Thr Asp Glu Ser Ser Lys

145 150 155 160

Asp Gly Pro Lys Gly Ile Phe Leu Gly Asp Gln Trp Arg Asp Ser Ala

165 170 175

Trp Gly Thr Ser Asp His Ser Val Ser Gln Pro Ile Met Val Gln Arg

180 185 190

Arg Pro Gly Gln Ser Phe His Val Asn Ser Glu Val Asn Ser Val Leu

195 200 205

Ser Pro Arg Ser Glu Ser Gly Gly Leu Gly Val Ser Met Val Glu Tyr

210 215 220

Val Leu Ser Ser Ser Pro Gly Asp Ser Cys Leu Arg Lys Gly Gly Phe

225 230 235 240

Gly Pro Arg Asp Ala Asp Ser Asp Glu Asn Asp Lys Gly Glu Lys Lys

245 250 255

Asn Lys Gly Thr Phe Asp Gly Asp Lys Leu Gly Asp Leu Lys Glu Glu

260 265 270

Gly Asp Val Met Asp Lys Thr Asn Gly Leu Pro Val Gln Asn Gly Ile

275 280 285

Asp Ala Asp Val Lys Asp Phe Ser Arg Thr Pro Gly Asn Cys Gln Asn

290 295 300

Ser Ala Asn Glu Val Asp Leu Leu Gly Pro Asn Gln Asn Gly Ser Glu

305 310 315 320

Gly Leu Ala Gln Leu Thr Ser Thr Asn Gly Ala Lys Pro Val Glu Asp

325 330 335

Phe Ser Asn Met Glu Ser Gln Ser Val Pro Leu Asp Pro Met Glu His

340 345 350

Val Gly Met Glu Pro Leu Gln Phe Asp Tyr Ser Gly Thr Gln Val Pro

355 360 365

Val Asp Ser Ala Ala Ala Thr Val Gly Leu Phe Asp Tyr Asn Ser Gln

370 375 380

Gln Gln Leu Phe Gln Arg Pro Asn Ala Leu Ala Val Gln Gln Leu Thr

385 390 395 400

Ala Ala Gln Gln Gln Gln Tyr Ala Leu Ala Ala Ala His Gln Pro His

405 410 415

Ile Gly Leu Ala Pro Ala Ala Phe Val Pro Asn Pro Tyr Ile Ile Ser

420 425 430

Ala Ala Pro Pro Gly Thr Asp Pro Tyr Thr Ala Gly Leu Ala Ala Ala

435 440 445

Ala Thr Leu Gly Pro Ala Val Val Pro His Gln Tyr Tyr Gly Val Thr

450 455 460

Pro Trp Gly Val Tyr Pro Ala Ser Leu Phe Gln Gln Gln Ala Ala Ala

465 470 475 480

Ala Ala Ala Ala Thr Asn Ser Ala Asn Gln Gln Thr Thr Pro Gln Ala

485 490 495

Gln Gln Gly Gln Gln Gln Val Leu Arg Gly Gly Ala Ser Gln Arg Pro

500 505 510

Leu Thr Pro Asn Gln Asn Gln Gln Gly Gln Gln Thr Asp Pro Leu Val

515 520 525

Ala Ala Ala Ala Val Asn Ser Ala Leu Ala Phe Gly Gln Gly Leu Ala

530 535 540

Ala Gly Met Pro Gly Tyr Pro Val Leu Ala Pro Ala Ala Tyr Tyr Asp

545 550 555 560

Gln Thr Gly Ala Leu Val Val Asn Ala Gly Ala Arg Asn Gly Leu Gly

565 570 575

Ala Pro Val Arg Leu Val Ala Pro Ala Pro Val Ile Ile Ser Ser Ser

580 585 590

Ala Ala Gln Ala Ala Val Ala Ala Ala Ala Ala Ser Ala Asn Gly Ala

595 600 605

Ala Gly Gly Leu Ala Gly Thr Thr Asn Gly Pro Phe Arg Pro Leu Gly

610 615 620

Thr Gln Gln Pro Gln Pro Gln Pro Gln Gln Gln Pro Asn Asn Asn Leu

625 630 635 640

Ala Ser Ser Ser Phe Tyr Gly Asn Asn Ser Leu Asn Ser Asn Ser Gln

645 650 655

Ser Ser Ser Leu Phe Ser Gln Gly Ser Ala Gln Pro Ala Asn Thr Ser

660 665 670

Leu Gly Phe Gly Ser Ser Ser Ser Leu Gly Ala Thr Leu Gly Ser Ala

675 680 685

Leu Gly Gly Phe Gly Thr Ala Val Ala Asn Ser Asn Thr Gly Ser Gly

690 695 700

Ser Arg Arg Asp Ser Leu Thr Gly Ser Ser Asp Leu Tyr Lys Arg Thr

705 710 715 720

Ser Ser Ser Leu Thr Pro Ile Gly His Ser Phe Tyr Asn Gly Leu Ser

725 730 735

Phe Ser Ser Ser Pro Gly Pro Val Gly Met Pro Leu Pro Ser Gln Gly

740 745 750

Pro Gly His Ser Gln Thr Pro Pro Pro Ser Leu Ser Ser His Gly Ser

755 760 765

Ser Ser Ser Leu Asn Leu Gly Gly Leu Thr Asn Gly Ser Gly Arg Tyr

770 775 780

Ile Ser Ala Ala Pro Gly Ala Glu Ala Lys Tyr Arg Ser Ala Ser Ser

785 790 795 800

Ala Ser Ser Leu Phe Ser Pro Ser Ser Thr Leu Phe Ser Ser Ser Arg

805 810 815

Leu Arg Tyr Gly Met Ser Asp Val Met Pro Ser Gly Arg Ser Arg Leu

820 825 830

Leu Glu Asp Phe Arg Asn Asn Arg Tyr Pro Asn Leu Gln Leu Arg Glu

835 840 845

Ile Ala Gly His Ile Met Glu Phe Ser Gln Asp Gln His Gly Ser Arg

850 855 860

Phe Ile Gln Leu Lys Leu Glu Arg Ala Thr Pro Ala Glu Arg Gln Leu

865 870 875 880

Val Phe Asn Glu Ile Leu Gln Ala Ala Tyr Gln Leu Met Val Asp Val

885 890 895

Phe Gly Asn Tyr Val Ile Gln Lys Phe Phe Glu Phe Gly Ser Leu Glu

900 905 910

Gln Lys Leu Ala Leu Ala Glu Arg Ile Arg Gly His Val Leu Ser Leu

915 920 925

Ala Leu Gln Met Tyr Gly Cys Arg Val Ile Gln Lys Ala Leu Glu Phe

930 935 940

Ile Pro Ser Asp Gln Gln Asn Glu Met Val Arg Glu Leu Asp Gly His

945 950 955 960

Val Leu Lys Cys Val Lys Asp Gln Asn Gly Asn His Val Val Gln Lys

965 970 975

Cys Ile Glu Cys Val Gln Pro Gln Ser Leu Gln Phe Ile Ile Asp Ala

980 985 990

Phe Lys Gly Gln Val Phe Ala Leu Ser Thr His Pro Tyr Gly Cys Arg

995 1000 1005

Val Ile Gln Arg Ile Leu Glu His Cys Leu Pro Asp Gln Thr Leu

1010 1015 1020

Pro Ile Leu Glu Glu Leu His Gln His Thr Glu Gln Leu Val Gln

1025 1030 1035

Asp Gln Tyr Gly Asn Tyr Val Ile Gln His Val Leu Glu His Gly

1040 1045 1050

Arg Pro Glu Asp Lys Ser Lys Ile Val Ala Glu Ile Arg Gly Asn

1055 1060 1065

Val Leu Val Leu Ser Gln His Lys Phe Ala Ser Asn Val Val Glu

1070 1075 1080

Lys Cys Val Thr His Ala Ser Arg Thr Glu Arg Ala Val Leu Ile

1085 1090 1095

Asp Glu Val Cys Thr Met Asn Asp Gly Pro His Ser Ala Leu Tyr

1100 1105 1110

Thr Met Met Lys Asp Gln Tyr Ala Asn Tyr Val Val Gln Lys Met

1115 1120 1125

Ile Asp Val Ala Glu Pro Gly Gln Arg Lys Ile Val Met His Lys

1130 1135 1140

Ile Arg Pro His Ile Ala Thr Leu Arg Lys Tyr Thr Tyr Gly Lys

1145 1150 1155

His Ile Leu Ala Lys Leu Glu Lys Tyr Tyr Met Lys Asn Gly Val

1160 1165 1170

Asp Leu Gly Pro Ile Cys Gly Pro Asn Gly Ile Ile

1175 1180 1185

<210> 48

<211> 22

<212> DNA

<213> Artificial sequence

<220>

<223> PUM1_forward primer

<400> 48

gccagcttgt cttcaatgaa at 22

<210> 49

<211> 21

<212> DNA

<213> Artificial sequence

<220>

<223> PUM1_reverse primer

<400> 49

caaagccagc ttctgttcaa g 21

<210> 50

<211> 24

<212> DNA

<213> Artificial sequence

<220>

<223> PUM1 probe

<400> 50

atccaccatg agttggtagg cagc 24

<210> 51

<211> 1921

<212> DNA

<213> Homo sapiens

<400> 51

ggcggaagtg acattatcaa cgcgcgccag gggttcagtg aggtcgggca ggttcgctgt 60

ggcgggcgcc tgggccgccg gctgtttaac ttcgcttccg ctggcccata gtgatctttg 120

cagtgaccca gcatcactgt ttcttggcgt gtgaagataa cccaaggaat tgaggaagtt 180

gctgagaaga gtgtgctgga gatgctctag gaaaaaattg aatagtgaga cgagttccag 240

cgcaagggtt tctggtttgc caagaagaaa gtgaacatca tggatcagaa caacagcctg 300

ccaccttacg ctcagggctt ggcctcccct cagggtgcca tgactcccgg aatccctatc 360

tttagtccaa tgatgcctta tggcactgga ctgaccccac agcctattca gaacaccaat 420

agtctgtcta ttttggaaga gcaacaaagg cagcagcagc aacaacaaca gcagcagcag 480

540

cagcagcagc agcagcagca acaggcagtg gcagctgcag ccgttcagca gtcaacgtcc 600

cagcaggcaa cacagggaac ctcaggccag gcaccacagc tcttccactc acagactctc 660

acaactgcac ccttgccggg caccactcca ctgtatccct cccccatgac tcccatgacc 720

cccatcactc ctgccacgcc agcttcggag agttctggga ttgtaccgca gctgcaaaat 780

attgtatcca cagtgaatct tggttgtaaa cttgacctaa agaccattgc acttcgtgcc 840

cgaaacgccg aatataatcc caagcggttt gctgcggtaa tcatgaggat aagagagcca 900

cgaaccacgg cactgatttt cagttctggg aaaatggtgt gcacaggagc caagagtgaa 960

gaacagtcca gactggcagc aagaaaatat gctagagttg tacagaagtt gggttttcca 1020

gctaagttct tggacttcaa gattcagaat atggtgggga gctgtgatgt gaagtttcct 1080

ataaggttag aaggccttgt gctcacccac caacaattta gtagttatga gccagagtta 1140

ttttcctggtt taatctacag aatgatcaaa cccagaattg ttctccttat ttttgtttct 1200

1260

atctacccta ttctaaaggg attcaggaag acgacgtaat ggctctcatg tacccttgcc 1320

tccccccaccc ccttcttttt tttttttttaa acaaatcagt ttgttttggt acctttaaat 1380

ggtggtgttg tgagaagatg gatgttgagt tgcagggtgt ggcaccaggt gatgcccttc 1440

tgtaagtgcc caccgcggga tgccgggaag gggcattatt tgtgcactga gaacaccgcg 1500

cagcgtgact gtgagttgct cataccgtgc tgctatctgg gcagcgctgc ccatttattt 1560

1620

taaagccacc tctataattg attggacttt ttaattttaa tgttttttccc catgaaccac 1680

agtttttata tttctaccag aaaagtaaaa atctttttta aaagtgttgt ttttctaatt 1740

tataactcct aggggtttatt tctgtgccag acacattcca cctctccagt attgcaggac 1800

agaatatatg tgttaatgaa aatgaatggc tgtacatatt ttttttctttc ttcagagtac 1860

tctgtacaat aaatgcagtt tataaaagtg ttagattgtt gttaaaaaaa aaaaaaaaaa 1920

a 1921

<210> 52

<211> 339

<212> PROTEIN

<213> Homo sapiens

<400> 52

Met Asp Gln Asn Asn Ser Leu Pro Pro Tyr Ala Gln Gly Leu Ala Ser

1 5 10 15

Pro Gln Gly Ala Met Thr Pro Gly Ile Pro Ile Phe Ser Pro Met Met

20 25 30

Pro Tyr Gly Thr Gly Leu Thr Pro Gln Pro Ile Gln Asn Thr Asn Ser

35 40 45

Leu Ser Ile Leu Glu Glu Gln Gln Arg Gln Gln Gln Gln Gln Gln Gln

50 55 60

Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln

65 70 75 80

Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Ala

85 90 95

Val Ala Ala Ala Ala Val Gln Gln Ser Thr Ser Gln Gln Ala Thr Gln

100 105 110

Gly Thr Ser Gly Gln Ala Pro Gln Leu Phe His Ser Gln Thr Leu Thr

115 120 125

Thr Ala Pro Leu Pro Gly Thr Thr Pro Leu Tyr Pro Ser Pro Met Thr

130 135 140

Pro Met Thr Pro Ile Thr Pro Ala Thr Pro Ala Ser Glu Ser Ser Gly

145 150 155 160

Ile Val Pro Gln Leu Gln Asn Ile Val Ser Thr Val Asn Leu Gly Cys

165 170 175

Lys Leu Asp Leu Lys Thr Ile Ala Leu Arg Ala Arg Asn Ala Glu Tyr

180 185 190

Asn Pro Lys Arg Phe Ala Ala Val Ile Met Arg Ile Arg Glu Pro Arg

195 200 205

Thr Thr Ala Leu Ile Phe Ser Ser Gly Lys Met Val Cys Thr Gly Ala

210 215 220

Lys Ser Glu Glu Gln Ser Arg Leu Ala Ala Arg Lys Tyr Ala Arg Val

225 230 235 240

Val Gln Lys Leu Gly Phe Pro Ala Lys Phe Leu Asp Phe Lys Ile Gln

245 250 255

Asn Met Val Gly Ser Cys Asp Val Lys Phe Pro Ile Arg Leu Glu Gly

260 265 270

Leu Val Leu Thr His Gln Gln Phe Ser Ser Tyr Glu Pro Glu Leu Phe

275 280 285

Pro Gly Leu Ile Tyr Arg Met Ile Lys Pro Arg Ile Val Leu Leu Ile

290 295 300

Phe Val Ser Gly Lys Val Val Leu Thr Gly Ala Lys Val Arg Ala Glu

305 310 315 320

Ile Tyr Glu Ala Phe Glu Asn Ile Tyr Pro Ile Leu Lys Gly Phe Arg

325 330 335

Lys Thr Thr

<210> 53

<211> 23

<212> DNA

<213> Artificial sequence

<220>

<223> TBP_forward primer

<400> 53

gccaagaaga aagtgaacat cat 23

<210> 54

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> TBP_reverse primer

<400> 54

atagggattc cgggagtcat 20

<210> 55

<211> 24

<212> DNA

<213> Artificial sequence

<220>

<223> TBP probe

<400> 55

tcagaacaac agcctgccac ctta 24

<210> 56

<211> 1852

<212> DNA

<213> Homo sapiens

<400> 56

accgccgaga ccgcgtccgc cccgcgagca cagagcctcg cctttgccga tccgccgccc 60

gtccacaccc gccgccagct caccatggat gatgatatcg ccgcgctcgt cgtcgacaac 120

ggctccggca tgtgcaaggc cggcttcgcg ggcgacgatg ccccccggggc cgtcttcccc 180

tccatcgtgg ggcgccccag gcaccagggc gtgatggtgg gcatgggtca gaaggattcc 240

tatgtgggcg acgaggccca gagcaagaga ggcatcctca ccctgaagta ccccatcgag 300

cacggcatcg tcaccaactg ggacgacatg gagaaaatct ggcaccacac cttctacaat 360

gagctgcgtg tggctcccga ggagcacccc gtgctgctga ccgaggcccc cctgaacccc 420

aaggccaacc gcgagaagat gacccagatc atgtttgaga ccttcaacac cccagccatg 480

tacgttgcta tccaggctgt gctatccctg tacgcctctg gccgtaccac tggcatcgtg 540

atggactccg gtgacggggt cacccacact gtgcccatct acgaggggta tgccctcccc 600

catgccatcc tgcgtctgga cctggctggc cgggacctga ctgactacct catgaagatc 660

ctcaccgagc gcggctacag cttcaccacc acggccgagc gggaaatcgt gcgtgacatt 720

aaggagaagc tgtgctacgt cgccctggac ttcgagcaag agatggccac ggctgcttcc 780

agctcctccc tggagaagag ctacgagctg cctgacggcc aggtcatcac cattggcaat 840

gagcggttcc gctgccctga ggcactcttc cagccttcct tcctgggcat ggagtcctgt 900

ggcatccacg aaactacctt caactccatc atgaagtgtg acgtggacat ccgcaaagac 960

ctgtacgcca acacagtgct gtctggcggc accaccatgt accctggcat tgccgacagg 1020

1080

cctgagcgca agtactccgt gtggatcggc ggctccatcc tggcctcgct gtccaccttc 1140

cagcagatgt ggatcagcaa gcaggagtat gacgagtccg gcccctccat cgtccaccgc 1200

aaatgcttct aggcggacta tgacttagtt gcgttacacc ctttcttgac aaaacctaac 1260

ttgcgcagaa aacaagatga gattggcatg gctttatttg ttttttttgt tttgttttgg 1320

tttttttttt ttttttggct tgactcagga tttaaaaact ggaacggtga aggtgacagc 1380

agtcggttgg agcgagcatc ccccaaagtt cacaatgtgg ccgaggactt tgattgcaca 1440

ttgttgtttt tttaatagtc attccaaata tgagatgcgt tgttacagga agtcccttgc 1500

catcctaaaa gccaccccac ttctctctaa ggagaatggc ccagtcctct cccaagtcca 1560

cacaggggag gtgatagcat tgctttcgtg taaattatgt aatgcaaaat ttttttaatc 1620

ttcgccttaa tactttttta ttttgtttta ttttgaatga tgagccttcg tgccccccct 1680

tccccctttt ttgtccccca acttgagatg tatgaaggct tttggtctcc ctgggagtgg 1740

gtggaggcag ccagggctta cctgtacact gacttgagac cagttgaata aaagtgcaca 1800

ccttaaaaat gaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aa 1852

<210> 57

<211> 375

<212> PROTEIN

<213> Homo sapiens

<400> 57

Met Asp Asp Asp Ile Ala Ala Leu Val Val Asp Asn Gly Ser Gly Met

1 5 10 15

Cys Lys Ala Gly Phe Ala Gly Asp Asp Ala Pro Arg Ala Val Phe Pro

20 25 30

Ser Ile Val Gly Arg Pro Arg His Gln Gly Val Met Val Gly Met Gly

35 40 45

Gln Lys Asp Ser Tyr Val Gly Asp Glu Ala Gln Ser Lys Arg Gly Ile

50 55 60

Leu Thr Leu Lys Tyr Pro Ile Glu His Gly Ile Val Thr Asn Trp Asp

65 70 75 80

Asp Met Glu Lys Ile Trp His His Thr Phe Tyr Asn Glu Leu Arg Val

85 90 95

Ala Pro Glu Glu His Pro Val Leu Leu Thr Glu Ala Pro Leu Asn Pro

100 105 110

Lys Ala Asn Arg Glu Lys Met Thr Gln Ile Met Phe Glu Thr Phe Asn

115 120 125

Thr Pro Ala Met Tyr Val Ala Ile Gln Ala Val Leu Ser Leu Tyr Ala

130 135 140

Ser Gly Arg Thr Thr Gly Ile Val Met Asp Ser Gly Asp Gly Val Thr

145 150 155 160

His Thr Val Pro Ile Tyr Glu Gly Tyr Ala Leu Pro His Ala Ile Leu

165 170 175

Arg Leu Asp Leu Ala Gly Arg Asp Leu Thr Asp Tyr Leu Met Lys Ile

180 185 190

Leu Thr Glu Arg Gly Tyr Ser Phe Thr Thr Thr Ala Glu Arg Glu Ile

195 200 205

Val Arg Asp Ile Lys Glu Lys Leu Cys Tyr Val Ala Leu Asp Phe Glu

210 215 220

Gln Glu Met Ala Thr Ala Ala Ser Ser Ser Ser Leu Glu Lys Ser Tyr

225 230 235 240

Glu Leu Pro Asp Gly Gln Val Ile Thr Ile Gly Asn Glu Arg Phe Arg

245 250 255

Cys Pro Glu Ala Leu Phe Gln Pro Ser Phe Leu Gly Met Glu Ser Cys

260 265 270

Gly Ile His Glu Thr Thr Phe Asn Ser Ile Met Lys Cys Asp Val Asp

275 280 285

Ile Arg Lys Asp Leu Tyr Ala Asn Thr Val Leu Ser Gly Gly Thr Thr

290 295 300

Met Tyr Pro Gly Ile Ala Asp Arg Met Gln Lys Glu Ile Thr Ala Leu

305 310 315 320

Ala Pro Ser Thr Met Lys Ile Lys Ile Ile Ala Pro Pro Glu Arg Lys

325 330 335

Tyr Ser Val Trp Ile Gly Gly Ser Ile Leu Ala Ser Leu Ser Thr Phe

340 345 350

Gln Gln Met Trp Ile Ser Lys Gln Glu Tyr Asp Glu Ser Gly Pro Ser

355 360 365

Ile Val His Arg Lys Cys Phe

370 375

<210> 58

<211> 18

<212> DNA

<213> Artificial sequence

<220>

<223> ACTB_forward primer

<400> 58

ccaaccgcga gaagatga 18

<210> 59

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> ACTB_reverse primer

<400> 59

ccagaggcgt acagggatag 20

<210> 60

<211> 23

<212> DNA

<213> Artificial sequence

<220>

<223> ACTB probe

<400> 60

ccatgtacgt tgctatccag gct 23

<210> 61

<211> 1229

<212> DNA

<213> Homo sapiens

<400> 61

gtctgacggg cgatggcgca gccaatagac aggagcgcta tccgcggttt ctgattggct 60

actttgttcg cattataaaa ggcacgcgcg ggcgcgaggc ccttctctcg ccaggcgtcc 120

tcgtggaagt gacatcgtct ttaaaccctg cgtggcaatc cctgacgcac cgccgtgatg 180

cccagggaag acagggcgac ctggaagtcc aactacttcc ttaagatcat ccaactattg 240

gatgattatc cgaaatgttt cattgtggga gcagacaatg tgggctccaa gcagatgcag 300

cagatccgca tgtcccttcg cgggaaggct gtggtgctga tgggcaagaa caccatgatg 360

cgcaaggcca tccgagggca cctggaaaac aacccagctc tggagaaact gctgcctcat 420

atccggggga atgtgggctt tgtgttcacc aaggaggacc tcactgagat cagggacatg 480

ttgctggcca ataaggtgcc agctgctgcc cgtgctggtg ccattgcccc atgtgaagtc 540

actgtgccag cccagaacac tggtctcggg cccgagaaga cctccttttt ccaggcttta 600

ggtatcacca ctaaaatctc caggggcacc attgaaatcc tgagtgatgt gcagctgatc 660

aagactggag acaaagtggg agccagcgaa gccacgctgc tgaacatgct caacatctcc 720

cccttctcct ttgggctggt catccagcag gtgttcgaca atggcagcat ctacaaccct 780

gaagtgcttg atatcacaga ggaaactctg cattctcgct tcctggaggg tgtccgcaat 840

gttgccagtg tctgtctgca gattggctac ccaactgttg catcagtacc ccattctatc 900

atcaacgggt acaaacgagt cctggccttg tctgtggaga cggattacac cttcccactt 960

gctgaaaagg tcaaggcctt cttggctgat ccatctgcct ttgtggctgc tgcccctgtg 1020

gctgctgcca ccacagctgc tcctgctgct gctgcagccc cagctaaggt tgaagccaag 1080

gaagagtcgg aggagtcgga cgaggatatg ggatttggtc tctttgacta atcaccaaaa 1140

agcaaccaac ttagccagtt ttatttgcaa aacaaggaaa taaaggctta cttctttaaa 1200

1229

<210> 62

<211> 317

<212> PROTEIN

<213> Homo sapiens

<400> 62

Met Pro Arg Glu Asp Arg Ala Thr Trp Lys Ser Asn Tyr Phe Leu Lys

1 5 10 15

Ile Ile Gln Leu Leu Asp Asp Tyr Pro Lys Cys Phe Ile Val Gly Ala

20 25 30

Asp Asn Val Gly Ser Lys Gln Met Gln Gln Ile Arg Met Ser Leu Arg

35 40 45

Gly Lys Ala Val Val Leu Met Gly Lys Asn Thr Met Met Arg Lys Ala

50 55 60

Ile Arg Gly His Leu Glu Asn Asn Pro Ala Leu Glu Lys Leu Leu Pro

65 70 75 80

His Ile Arg Gly Asn Val Gly Phe Val Phe Thr Lys Glu Asp Leu Thr

85 90 95

Glu Ile Arg Asp Met Leu Leu Ala Asn Lys Val Pro Ala Ala Ala Arg

100 105 110

Ala Gly Ala Ile Ala Pro Cys Glu Val Thr Val Pro Ala Gln Asn Thr

115 120 125

Gly Leu Gly Pro Glu Lys Thr Ser Phe Phe Gln Ala Leu Gly Ile Thr

130 135 140

Thr Lys Ile Ser Arg Gly Thr Ile Glu Ile Leu Ser Asp Val Gln Leu

145 150 155 160

Ile Lys Thr Gly Asp Lys Val Gly Ala Ser Glu Ala Thr Leu Leu Asn

165 170 175

Met Leu Asn Ile Ser Pro Phe Ser Phe Gly Leu Val Ile Gln Gln Val

180 185 190

Phe Asp Asn Gly Ser Ile Tyr Asn Pro Glu Val Leu Asp Ile Thr Glu

195 200 205

Glu Thr Leu His Ser Arg Phe Leu Glu Gly Val Arg Asn Val Ala Ser

210 215 220

Val Cys Leu Gln Ile Gly Tyr Pro Thr Val Ala Ser Val Pro His Ser

225 230 235 240

Ile Ile Asn Gly Tyr Lys Arg Val Leu Ala Leu Ser Val Glu Thr Asp

245 250 255

Tyr Thr Phe Pro Leu Ala Glu Lys Val Lys Ala Phe Leu Ala Asp Pro

260 265 270

Ser Ala Phe Val Ala Ala Ala Pro Val Ala Ala Ala Thr Thr Ala Ala

275 280 285

Pro Ala Ala Ala Ala Ala Pro Ala Lys Val Glu Ala Lys Glu Glu Ser

290 295 300

Glu Glu Ser Asp Glu Asp Met Gly Phe Gly Leu Phe Asp

305 310 315

<210> 63

<211> 317

<212> PROTEIN

<213> Homo sapiens

<400> 63

Met Pro Arg Glu Asp Arg Ala Thr Trp Lys Ser Asn Tyr Phe Leu Lys

1 5 10 15

Ile Ile Gln Leu Leu Asp Asp Tyr Pro Lys Cys Phe Ile Val Gly Ala

20 25 30

Asp Asn Val Gly Ser Lys Gln Met Gln Gln Ile Arg Met Ser Leu Arg

35 40 45

Gly Lys Ala Val Val Leu Met Gly Lys Asn Thr Met Met Arg Lys Ala

50 55 60

Ile Arg Gly His Leu Glu Asn Asn Pro Ala Leu Glu Lys Leu Leu Pro

65 70 75 80

His Ile Arg Gly Asn Val Gly Phe Val Phe Thr Lys Glu Asp Leu Thr

85 90 95

Glu Ile Arg Asp Met Leu Leu Ala Asn Lys Val Pro Ala Ala Ala Arg

100 105 110

Ala Gly Ala Ile Ala Pro Cys Glu Val Thr Val Pro Ala Gln Asn Thr

115 120 125

Gly Leu Gly Pro Glu Lys Thr Ser Phe Phe Gln Ala Leu Gly Ile Thr

130 135 140

Thr Lys Ile Ser Arg Gly Thr Ile Glu Ile Leu Ser Asp Val Gln Leu

145 150 155 160

Ile Lys Thr Gly Asp Lys Val Gly Ala Ser Glu Ala Thr Leu Leu Asn

165 170 175

Met Leu Asn Ile Ser Pro Phe Ser Phe Gly Leu Val Ile Gln Gln Val

180 185 190

Phe Asp Asn Gly Ser Ile Tyr Asn Pro Glu Val Leu Asp Ile Thr Glu

195 200 205

Glu Thr Leu His Ser Arg Phe Leu Glu Gly Val Arg Asn Val Ala Ser

210 215 220

Val Cys Leu Gln Ile Gly Tyr Pro Thr Val Ala Ser Val Pro His Ser

225 230 235 240

Ile Ile Asn Gly Tyr Lys Arg Val Leu Ala Leu Ser Val Glu Thr Asp

245 250 255

Tyr Thr Phe Pro Leu Ala Glu Lys Val Lys Ala Phe Leu Ala Asp Pro

260 265 270

Ser Ala Phe Val Ala Ala Ala Pro Val Ala Ala Ala Thr Thr Ala Ala

275 280 285

Pro Ala Ala Ala Ala Ala Pro Ala Lys Val Glu Ala Lys Glu Glu Ser

290 295 300

Glu Glu Ser Asp Glu Asp Met Gly Phe Gly Leu Phe Asp

305 310 315

<210> 64

<211> 18

<212> DNA

<213> Artificial sequence

<220>

<223> RPLP0_forward primer

<400> 64

taaaccctgc gtggcaat 18

<210> 65

<211> 27

<212> DNA

<213> Artificial sequence

<220>

<223> RPLP0_reverse primer

<400> 65

acatttcgga taatcatcca atagttg 27

<210> 66

<211> 24

<212> DNA

<213> Artificial sequence

<220>

<223> RPLP0 probe

<400> 66

aagtagttgg acttccaggt cgcc 24

<210> 67

<211> 2458

<212> DNA

<213> Homo sapiens

<400> 67

cagaagaagg cagcgcccaa ggcgcatgcg cagcggtcac tcccgctgta tattaaggcg 60

ccggcgatcg cggcctgagg ctgctcccgg acaagggcaa cgagcgtttc gtttggactt 120

ctcgacttga gtgcccgcct ccttcgccgc cgcctctgca gtcctcagcg cagttatgcc 180

cagttcttcc cgctgtgggg acacgaccac ggaggaatcc ttgcttcagg gactcgggac 240

cctgctggac cccttcctcg ggtttaggggg atgtggggac caggagaaag tcaggatccc 300

taagagtctt ccctgcctgg atggatgagt ggcttcttct ccacctagat tctttccaca 360

ggagccagca tacttcctga acatggagag tgttgttcgc cgctgcccat tcttatcccg 420

agtcccccag gcctttctgc agaaagcagg caaatctctg ttgttctatg cccaaaactg 480

ccccaagatg atggaagttg gggccaagcc agcccctcgg gcattgtcca ctgcagcagt 540

acactaccaa cagatcaaag aaacccctcc ggccagtgag aaagacaaaa ctgctaaggc 600

caaggtccaa cagactcctg atggatccca gcagagtcca gatggcacac agcttccgtc 660

tggacaccc ttgcctgcca caagccaggg cactgcaagc aaatgccctt tcctggcagc 720

acagatgaat cagagaggca gcagtgtctt ctgcaaagcc agtcttgagc ttcaggagga 780

tgtgcaggaa atgaatgccg tgaggaaaga ggttgctgaa acctcagcag gccccagtgt 840

ggttagtgtg aaaaccgatg gaggggatcc cagtggactg ctgaagaact tccaggacat 900

catgcaaaag caaagaccag aaagagtgtc tcatcttctt caagataact tgccaaaatc 960

1020

ccacacctat cgagttttta aaactgtgaa ccggcgagca cacatcttcc ccatggcaga 1080

tgactattca gactccctca tcaccaaaaa gcaagtgtca gtctggtgca gtaatgacta 1140

cctaggaatg agtcgccacc cacgggtgtg tggggcagtt atggacactt tgaaacaaca 1200

tggtgctggg gcaggtggta ctagaaatat ttctggaact agtaaattcc atgtggactt 1260

agagcgggag ctggcagacc tccatgggaa agatgccgca ctcttgtttt cctcgtgctt 1320

tgtggccaat gactcaaccc tcttcaccct ggctaagatg atgccaggct gtgagattta 1380

ctctgattct gggaaccatg ccctccatgat ccaagggatt cgaaacagcc gagtgccaaa 1440

gtacatcttc cgccacaatg atgtcagcca cctcagagaa ctgctgcaaa gatctgaccc 1500

ctcagtcccc aagattgtgg catttgaaac tgtccattca atggatgggg cggtgtgccc 1560

actggaagag ctgtgtgatg tggcccatga gtttggagca atcaccttcg tggatgaggt 1620

ccacgcagtg gggctttatg gggctcgagg cggagggatt ggggatcggg atggagtcat 1680

gccaaaaatg gacatcattt ctggaacact tggcaaagcc tttggttgtg ttggagggta 1740

catcgccagc acgagttctc tgattgacac cgtacggtcc tatgctgctg gcttcatctt 1800

caccacctct ctgccaccca tgctgctggc tggagccctg gagtctgtgc ggatcctgaa 1860

gagcgctgag ggacgggtgc ttcgccgcca gcaccagcgc aacgtcaaac tcatgagaca 1920

gatgctaatg gatgccggcc tccctgttgt ccactgcccc agccacatca tccctgtgcg 1980

ggttgcagat gctgctaaaa acacagaagt ctgtgatgaa ctaatgagca gacataacat 2040

ctacgtgcaa gcaatcaatt accctacggt gccccggggga gaagagctcc tacggattgc 2100

ccccacccct caccacacac ccgatgat gaactacttc cttgagaatc tgctagtcac 2160

atggaagcaa gtggggctgg aactgaagcc tcattcctca gctgagtgca acttctgcag 2220

2280

caagttggta tctgctcagg cctgagcatg acctcaatta tttcacttaa ccccaggcca 2340

ttatcatatc cagatggtct tcagagttgt ctttatatgt gaattaagtt atattaaatt 2400

ttaatctata gtaaaaacat agtcctggaa ataaattctt gcttaaatgg tgaaaaaa 2458

<210> 68

<211> 2281

<212> DNA

<213> Homo sapiens

<400> 68

cagaagaagg cagcgcccaa ggcgcatgcg cagcggtcac tcccgctgta tattaaggcg 60

ccggcgatcg cggcctgagg ctgctcccgg acaagggcaa cgagcgtttc gtttggactt 120

ctcgacttga gtgcccgcct ccttcgccgc cgcctctgca gtcctcagcg cagtctttcc 180

acaggagcca gcatacttcc tgaacatgga gagtgttgtt cgccgctgcc cattcttatc 240

ccgagtcccc caggcctttc tgcagaaagc aggcaaatct ctgttgttct atgcccaaaa 300

ctgccccaag atgatggaag ttggggccaa gccagcccct cgggcattgt ccactgcagc 360

agtacactac caacagatca aagaaacccc tccggccagt gagaaagaca aaactgctaa 420

ggccaaggtc caacagactc ctgatggatc ccagcagagt ccagatggca cacagcttcc 480

gtctggacac cccttgcctg ccacaagcca gggcactgca agcaaatgcc ctttcctggc 540

agcacagatg aatcagagag gcagcagtgt cttctgcaaa gccagtcttg agcttcagga 600

ggatgtgcag gaaatgaatg ccgtgaggaa agaggttgct gaaacctcag caggccccag 660

tgtggttagt gtgaaaaccg atggagggga tcccagtgga ctgctgaaga acttccagga 720

catcatgcaa aagcaaagac cagaaagagt gtctcatctt cttcaagata acttgccaaa 780

atctgtttcc acttttcagt atgatcgttt ctttgagaaa aaaattgatg agaaaaagaa 840

tgaccacacc tatcgagttt ttaaaactgt gaaccggcga gcacacatct tccccatggc 900

agatgactat tcagactccc tcatcaccaa aaagcaagtg tcagtctggt gcagtaatga 960

ctacctagga atgagtcgcc acccacggggt gtgtggggca gttatggaca ctttgaaaca 1020

acatggtgct ggggcaggtg gtactagaaa tatttctgga actagtaaat tccatgtgga 1080

cttagagcgg gagctggcag acctccatgg gaaagatgcc gcactcttgt tttcctcgtg 1140

ctttgtggcc aatgactcaa ccctcttcac cctggctaag atgatgccag gctgtgagat 1200

ttactctgat tctgggaacc atgcctccat gatccaaggg attcgaaaca gccgagtgcc 1260

aaagtacatc ttccgccaca atgatgtcag ccacctcaga gaactgctgc aaagatctga 1320

cccctcagtc cccaagattg tggcatttga aactgtccat tcaatggatg gggcggtgtg 1380

cccactggaa gagctgtgtg atgtggccca tgagtttgga gcaatcacct tcgtggatga 1440

ggtccacgca gtggggcttt atggggctcg aggcggaggg attggggatc gggatggagt 1500

catgccaaaa atggacatca tttctggaac acttggcaaa gcctttggtt gtgttgggagg 1560

gtacatcgcc agcacgagtt ctctgattga caccgtacgg tcctatgctg ctggcttcat 1620

cttcaccacc tctctgccac ccatgctgct ggctggagcc ctggagtctg tgcggatcct 1680

gaagagcgct gaggacggg tgcttcgccg ccagcaccag cgcaacgtca aactcatgag 1740

acagatgcta atggatgccg gcctccctgt tgtccactgc cccagccaca tcatccctgt 1800

gcgggttgca gatgctgcta aaaacacaga agtctgtgat gaactaatga gcagacataa 1860

catctacgtg caagcaatca attaccctac ggtgccccgg ggagaagagc tcctacggat 1920

tgccccccacc cctcaccaca caccccagat gatgaactac ttccttgaga atctgctagt 1980

cacatggaag caagtggggc tggaactgaa gcctcattcc tcagctgagt gcaacttctg 2040

caggaggcca ctgcattttg aagtgatgag tgaaagagag aagtcctatt tctcaggctt 2100

gagcaagttg gtatctgctc aggcctgagc atgacctcaa ttatttcact taaccccagg 2160

ccattatcat atccagatgg tcttcagagt tgtctttata tgtgaattaa gttatattaa 2220

attttaatct atagtaaaaa catagtcctg gaaataaatt cttgcttaaa tggtgaaaaa 2280

a 2281

<210> 69

<211> 640

<212> PROTEIN

<213> Homo sapiens

<400> 69

Met Glu Ser Val Val Arg Arg Cys Pro Phe Leu Ser Arg Val Pro Gln

1 5 10 15

Ala Phe Leu Gln Lys Ala Gly Lys Ser Leu Leu Phe Tyr Ala Gln Asn

20 25 30

Cys Pro Lys Met Met Glu Val Gly Ala Lys Pro Ala Pro Arg Ala Leu

35 40 45

Ser Thr Ala Ala Val His Tyr Gln Gln Ile Lys Glu Thr Pro Pro Ala

50 55 60

Ser Glu Lys Asp Lys Thr Ala Lys Ala Lys Val Gln Gln Thr Pro Asp

65 70 75 80

Gly Ser Gln Gln Ser Pro Asp Gly Thr Gln Leu Pro Ser Gly His Pro

85 90 95

Leu Pro Ala Thr Ser Gln Gly Thr Ala Ser Lys Cys Pro Phe Leu Ala

100 105 110

Ala Gln Met Asn Gln Arg Gly Ser Ser Val Phe Cys Lys Ala Ser Leu

115 120 125

Glu Leu Gln Glu Asp Val Gln Glu Met Asn Ala Val Arg Lys Glu Val

130 135 140

Ala Glu Thr Ser Ala Gly Pro Ser Val Val Ser Val Lys Thr Asp Gly

145 150 155 160

Gly Asp Pro Ser Gly Leu Leu Lys Asn Phe Gln Asp Ile Met Gln Lys

165 170 175

Gln Arg Pro Glu Arg Val Ser His Leu Leu Gln Asp Asn Leu Pro Lys

180 185 190

Ser Val Ser Thr Phe Gln Tyr Asp Arg Phe Phe Glu Lys Lys Ile Asp

195 200 205

Glu Lys Lys Asn Asp His Thr Tyr Arg Val Phe Lys Thr Val Asn Arg

210 215 220

Arg Ala His Ile Phe Pro Met Ala Asp Asp Tyr Ser Asp Ser Leu Ile

225 230 235 240

Thr Lys Lys Gln Val Ser Val Trp Cys Ser Asn Asp Tyr Leu Gly Met

245 250 255

Ser Arg His Pro Arg Val Cys Gly Ala Val Met Asp Thr Leu Lys Gln

260 265 270

His Gly Ala Gly Ala Gly Gly Thr Arg Asn Ile Ser Gly Thr Ser Lys

275 280 285

Phe His Val Asp Leu Glu Arg Glu Leu Ala Asp Leu His Gly Lys Asp

290 295 300

Ala Ala Leu Leu Phe Ser Ser Cys Phe Val Ala Asn Asp Ser Thr Leu

305 310 315 320

Phe Thr Leu Ala Lys Met Met Pro Gly Cys Glu Ile Tyr Ser Asp Ser

325 330 335

Gly Asn His Ala Ser Met Ile Gln Gly Ile Arg Asn Ser Arg Val Pro

340 345 350

Lys Tyr Ile Phe Arg His Asn Asp Val Ser His Leu Arg Glu Leu Leu

355 360 365

Gln Arg Ser Asp Pro Ser Val Pro Lys Ile Val Ala Phe Glu Thr Val

370 375 380

His Ser Met Asp Gly Ala Val Cys Pro Leu Glu Glu Leu Cys Asp Val

385 390 395 400

Ala His Glu Phe Gly Ala Ile Thr Phe Val Asp Glu Val His Ala Val

405 410 415

Gly Leu Tyr Gly Ala Arg Gly Gly Gly Ile Gly Asp Arg Asp Gly Val

420 425 430

Met Pro Lys Met Asp Ile Ile Ser Gly Thr Leu Gly Lys Ala Phe Gly

435 440 445

Cys Val Gly Gly Tyr Ile Ala Ser Thr Ser Ser Leu Ile Asp Thr Val

450 455 460

Arg Ser Tyr Ala Ala Gly Phe Ile Phe Thr Thr Ser Leu Pro Pro Met

465 470 475 480

Leu Leu Ala Gly Ala Leu Glu Ser Val Arg Ile Leu Lys Ser Ala Glu

485 490 495

Gly Arg Val Leu Arg Arg Gln His Gln Arg Asn Val Lys Leu Met Arg

500 505 510

Gln Met Leu Met Asp Ala Gly Leu Pro Val Val His Cys Pro Ser His

515 520 525

Ile Ile Pro Val Arg Val Ala Asp Ala Ala Lys Asn Thr Glu Val Cys

530 535 540

Asp Glu Leu Met Ser Arg His Asn Ile Tyr Val Gln Ala Ile Asn Tyr

545 550 555 560

Pro Thr Val Pro Arg Gly Glu Glu Leu Leu Arg Ile Ala Pro Thr Pro

565 570 575

His His Thr Pro Gln Met Met Asn Tyr Phe Leu Glu Asn Leu Leu Val

580 585 590

Thr Trp Lys Gln Val Gly Leu Glu Leu Lys Pro His Ser Ser Ala Glu

595 600 605

Cys Asn Phe Cys Arg Arg Pro Leu His Phe Glu Val Met Ser Glu Arg

610 615 620

Glu Lys Ser Tyr Phe Ser Gly Leu Ser Lys Leu Val Ser Ala Gln Ala

625 630 635 640

<210> 70

<211> 640

<212> PROTEIN

<213> Homo sapiens

<400> 70

Met Glu Ser Val Val Arg Arg Cys Pro Phe Leu Ser Arg Val Pro Gln

1 5 10 15

Ala Phe Leu Gln Lys Ala Gly Lys Ser Leu Leu Phe Tyr Ala Gln Asn

20 25 30

Cys Pro Lys Met Met Glu Val Gly Ala Lys Pro Ala Pro Arg Ala Leu

35 40 45

Ser Thr Ala Ala Val His Tyr Gln Gln Ile Lys Glu Thr Pro Pro Ala

50 55 60

Ser Glu Lys Asp Lys Thr Ala Lys Ala Lys Val Gln Gln Thr Pro Asp

65 70 75 80

Gly Ser Gln Gln Ser Pro Asp Gly Thr Gln Leu Pro Ser Gly His Pro

85 90 95

Leu Pro Ala Thr Ser Gln Gly Thr Ala Ser Lys Cys Pro Phe Leu Ala

100 105 110

Ala Gln Met Asn Gln Arg Gly Ser Ser Val Phe Cys Lys Ala Ser Leu

115 120 125

Glu Leu Gln Glu Asp Val Gln Glu Met Asn Ala Val Arg Lys Glu Val

130 135 140

Ala Glu Thr Ser Ala Gly Pro Ser Val Val Ser Val Lys Thr Asp Gly

145 150 155 160

Gly Asp Pro Ser Gly Leu Leu Lys Asn Phe Gln Asp Ile Met Gln Lys

165 170 175

Gln Arg Pro Glu Arg Val Ser His Leu Leu Gln Asp Asn Leu Pro Lys

180 185 190

Ser Val Ser Thr Phe Gln Tyr Asp Arg Phe Phe Glu Lys Lys Ile Asp

195 200 205

Glu Lys Lys Asn Asp His Thr Tyr Arg Val Phe Lys Thr Val Asn Arg

210 215 220

Arg Ala His Ile Phe Pro Met Ala Asp Asp Tyr Ser Asp Ser Leu Ile

225 230 235 240

Thr Lys Lys Gln Val Ser Val Trp Cys Ser Asn Asp Tyr Leu Gly Met

245 250 255

Ser Arg His Pro Arg Val Cys Gly Ala Val Met Asp Thr Leu Lys Gln

260 265 270

His Gly Ala Gly Ala Gly Gly Thr Arg Asn Ile Ser Gly Thr Ser Lys

275 280 285

Phe His Val Asp Leu Glu Arg Glu Leu Ala Asp Leu His Gly Lys Asp

290 295 300

Ala Ala Leu Leu Phe Ser Ser Cys Phe Val Ala Asn Asp Ser Thr Leu

305 310 315 320

Phe Thr Leu Ala Lys Met Met Pro Gly Cys Glu Ile Tyr Ser Asp Ser

325 330 335

Gly Asn His Ala Ser Met Ile Gln Gly Ile Arg Asn Ser Arg Val Pro

340 345 350

Lys Tyr Ile Phe Arg His Asn Asp Val Ser His Leu Arg Glu Leu Leu

355 360 365

Gln Arg Ser Asp Pro Ser Val Pro Lys Ile Val Ala Phe Glu Thr Val

370 375 380

His Ser Met Asp Gly Ala Val Cys Pro Leu Glu Glu Leu Cys Asp Val

385 390 395 400

Ala His Glu Phe Gly Ala Ile Thr Phe Val Asp Glu Val His Ala Val

405 410 415

Gly Leu Tyr Gly Ala Arg Gly Gly Gly Ile Gly Asp Arg Asp Gly Val

420 425 430

Met Pro Lys Met Asp Ile Ile Ser Gly Thr Leu Gly Lys Ala Phe Gly

435 440 445

Cys Val Gly Gly Tyr Ile Ala Ser Thr Ser Ser Leu Ile Asp Thr Val

450 455 460

Arg Ser Tyr Ala Ala Gly Phe Ile Phe Thr Thr Ser Leu Pro Pro Met

465 470 475 480

Leu Leu Ala Gly Ala Leu Glu Ser Val Arg Ile Leu Lys Ser Ala Glu

485 490 495

Gly Arg Val Leu Arg Arg Gln His Gln Arg Asn Val Lys Leu Met Arg

500 505 510

Gln Met Leu Met Asp Ala Gly Leu Pro Val Val His Cys Pro Ser His

515 520 525

Ile Ile Pro Val Arg Val Ala Asp Ala Ala Lys Asn Thr Glu Val Cys

530 535 540

Asp Glu Leu Met Ser Arg His Asn Ile Tyr Val Gln Ala Ile Asn Tyr

545 550 555 560

Pro Thr Val Pro Arg Gly Glu Glu Leu Leu Arg Ile Ala Pro Thr Pro

565 570 575

His His Thr Pro Gln Met Met Asn Tyr Phe Leu Glu Asn Leu Leu Val

580 585 590

Thr Trp Lys Gln Val Gly Leu Glu Leu Lys Pro His Ser Ser Ala Glu

595 600 605

Cys Asn Phe Cys Arg Arg Pro Leu His Phe Glu Val Met Ser Glu Arg

610 615 620

Glu Lys Ser Tyr Phe Ser Gly Leu Ser Lys Leu Val Ser Ala Gln Ala

625 630 635 640

<210> 71

<211> 17

<212> DNA

<213> Artificial sequence

<220>

<223> ALAS-1_forward primer

<400> 71

agccacatca tccctgt 17

<210> 72

<211> 22

<212> DNA

<213> Artificial sequence

<220>

<223> ALAS-1_reverse primer

<400> 72

cgtagatgtt atgtctgctc at 22

<210> 73

<211> 22

<212> DNA

<213> Artificial sequence

<220>

<223> ALAS-1 probe

<400> 73

tttagcagca tctgcaaccc gc 22

<210> 74

<211> 4543

<212> DNA

<213> Homo sapiens

<400> 74

atgcgtacgc tcgctggccc cgcccctag cgccgcggtc ggagccattt cgccgattcc 60

tccatgcgag ttgctgtgcg tttctctgtt gtctcggtag aaggccagag tcacacacgg 120

tcctaagagc tgggcaccag gaagcgaagg ctgatctgaa gaagacactt gaatcatggg 180

tgacgttaaa aattttctgt atgcctggtg tggcaaaagg aagatgaccc catcctatga 240

aattagagca gtggggaaca aaaacaggca gaaattcatg tgtgaggttc aggtggaagg 300

ttataattac actggcatgg gaaattccac caataaaaaa gatgcacaaa gcaatgctgc 360

cagagacttt gttaactatt tggttcgaat aaatgaaata aagagtgaag aagttccagc 420

ttttggggta gcatctccgc ccccacttac tgatactcct gacactacag caaatgctga 480

aggagattta ccaacaacca tgggaggacc tcttcctcca catctggctc tcaaagcaga 540

aaataattct gaggtaggggg cctctggcta tggtgttcct gggcccacct gggaccgagg 600

agccaacttg aaggattact actcaagaaa ggaagaacaa gaagtgcaag cgactctaga 660

atcagaagaa gtggatttaa atgctgggct tcatggaaac tggaccttgg aaaatgctaa 720

agctcgtcta aaccaatatt ttcagaaaga aaagatccaa ggagaatata agtacaccca 780

agtgggtcct gatcacaaca ggagctttat tgcagaaatg accatttata tcaagcagct 840

gggcagaagg atttttgcac gagaacatgg atcaaataag aaattggcag cacagtcctg 900

tgccctgtca cttgtcagac aactgtacca tcttggagtg gttgaagctt actccggact 960

tacaaagaag aaggaaggag agacagtgga gccttacaaa gtaaacctct ctcaagattt 1020

agagcatcag ctgcaaaaca tcattcaaga gctaaatctt gagattttgc ccccgcctga 1080

agatccttct gtgccagttg cactcaacat tggcaaattg gctcagttcg aaccatctca 1140

gcgacaaaac caagtgggtg tggttccttg gtcacctcca caatccaact ggaatccttg 1200

gactagtagc aacattgatg aggggcctct ggcttttgct actccagagc aaataagcat 1260

ggacctcaag aatgaattga tgtaccagtt ggaacaggat catgatttgc aagcaatctt 1320

gcaggagaga gagttactgc ctgtgaagaa atttgaaagt gagattctgg aagcaatcag 1380

ccaaaattca gttgtcatta ttagaggggc tactggatgt gggaaaacca cacaggttcc 1440

ccagttcatt ctagatgact ttatccagaa tgaccgagca gcagagtgta acatcgtagt 1500

aactcagccc agaagaatca gtgcggtttc tgtggcagag cgagttgcat ttgaaagagg 1560

agaagagcct ggaaaaagct gtggctacag cgttcgattt gagtctatac ttcctcgtcc 1620

tcatgccagt ataatgtttt gtactgtagg tgtgctcctg agaaaattag aagcaggcat 1680

tcgaggaatc agtcatgtaa ttgtagatga aatacatgaa agagatatta atactgactt 1740

ccttttggta gtactgcgtg atgttgttca ggcttatcct gaagttcgca ttgttcttat 1800

gtctgctact attgatacca gcatgttttg tgaatatttc ttcaattgcc ccatcattga 1860

agtttatggg aggacttacc cagttcaaga atattttctg gaagactgca ttcagatgac 1920

ccactttgtt cctccaccaa aagacaaaaa gaagaaggat aaggatgatg atggtggtga 1980

ggatgatgat gcaaattgca acttgatctg tggtgatgaa tatggtccag aaacaaggtt 2040

gagcatgtct caattgaacg aaaaggaaac tccttttgaa ctcatcgagg ctctacttaa 2100

gtacattgaa acccttaatg ttcctggagc tgtgttggtt ttttgcctg gctggaatct 2160

gatttatact atgcagaagc atttggaaat gaatccacat tttggaagcc atcggtatca 2220

gattctaccc ctgcattctc agattcctcg agaggaacag cgcaaagtgt ttgatccagt 2280

accagttgga gtaaccaagg ttattttgtc cacaaatatt gctgaaacaa gcattaccat 2340

aaacgatgtt gtttatgtca ttgactcctg caagcagaaa gtgaaactct tcactgctca 2400

caacaatatg accaactatg ctaccgtatg ggcatcaaaa acaaaccttg agcaacggaa 2460

agggcgagct ggccgagtac ggcctggatt ctgctttcac ctgtgcagcc gagctcgttt 2520

tgagagactt gaaacccaca tgacaccaga gatgttccga acaccattgc atgaaattgc 2580

tcttagcata aaacttctgc gtctaggagg aattggccaa tttctggcca aagcaattga 2640

acctccccct ttggatgctg tgattgaagc agaacacact cttagagagc ttgatgcatt 2700

agatgccaat gatgagttga ctcctttggg acgaatcctg gctaaactcc ccattgagcc 2760

tcgttttggc aaaatgatga taatggggtg tattttctac gtgggagatg ctatctgtac 2820

cattgctgct gctacctgct ttccagagcc tttcatcaat gaaggaaagc ggctgggcta 2880

tatccatcga aattttgctg gaaacagatt ttctgatcac gtagcccttt tatcagtatt 2940

ccaagcctgg gatgatgcta gaatgggtgg agaagaagca gagatacgtt tttgtgagca 3000

aatatggcta cactaagaat gacctgggaa gccaaagttc agctcaaaga 3060

3120

tggaccagat aataatttgg atgttgttat ctccctcctg gcctttggtg tgtaccccaa 3180

tgtatgctat cataaggaaa agaggaagat tctcaccact gaagggcgta atgcacttat 3240

ccacaaatca tctgttaatt gtccttttag tagccaagac atgaagtacc catctccctt 3300

3360

cacccccctg cagttgcttc tctttgcctc caagaaagtc caatctgatg ggcagattgt 3420

gcttgtagat gactggatta aactgcaaat atctcatgaa gctgctgcct gtatcactgg 3480

tctccgggca gccatgggagg ctttggttgt tgaagtaacc aaacaacctg ctatcatcag 3540

ccagttggac cccgtaaatg aacgtatgct gaacatgatc cgtcagatct ctagaccctc 3600

agctgctggt atcaacctta tgattggcag tacacggtat ggagatggtc cacgtcctcc 3660

caagatggcc cgatacgaca atggaagcgg atatagaagg ggaggttcta gttacagtgg 3720

tggaggctat ggcggtggct atagcagtgg aggctatggt agcgggaggct atggtggcag 3780

3840

ccgaggtggc tttagaggca actctggagg agactacaga gggcctagtg gaggctacag 3900

aggatctggg ggattccagc gaggaggtgg taggggggcc tatggaactg gctactttgg 3960

acagggaaga ggaggtggcg gctattaaaa cttggttatg tcagttcctg tgtgtagaca 4020

gtaaggaaaa aaaggcatgc tatgtgttac gtgttttttc cagtatgttt atttgccacc 4080

aaaaagtaaa tgcattttca cccattctgt ggttcattgt agtttaagga aaccaagcat 4140

atagatgcat tagtgatttt gtttatatta tgtaaaatat aacgatctct taaaaatacc 4200

acagtttgta ttttttcttt aaggagtaaa gatttgcctt taaataactt ggtattttcc 4260

tggctttcgt ttaatacaat agaaaataaa gtattacacc gaatacttgc cgtgtagttt 4320

4380

tgtcagagaa gttgtaccct gtttcaaaag tatactaagt gatactactt gtaatagaat 4440

aaatcatctt ggaattgaat tgttaccttt tgaagtaaat actggcaagt gcacaagcca 4500

cataaacctg aataaaactt ttgacctagg gttgaaaaaa aaa 4543

<210> 75

<211> 1270

<212> PROTEIN

<213> Homo sapiens

<400> 75

Met Gly Asp Val Lys Asn Phe Leu Tyr Ala Trp Cys Gly Lys Arg Lys

1 5 10 15

Met Thr Pro Ser Tyr Glu Ile Arg Ala Val Gly Asn Lys Asn Arg Gln

20 25 30

Lys Phe Met Cys Glu Val Gln Val Glu Gly Tyr Asn Tyr Thr Gly Met

35 40 45

Gly Asn Ser Thr Asn Lys Lys Asp Ala Gln Ser Asn Ala Ala Arg Asp

50 55 60

Phe Val Asn Tyr Leu Val Arg Ile Asn Glu Ile Lys Ser Glu Glu Val

65 70 75 80

Pro Ala Phe Gly Val Ala Ser Pro Pro Pro Leu Thr Asp Thr Pro Asp

85 90 95

Thr Thr Ala Asn Ala Glu Gly Asp Leu Pro Thr Thr Met Gly Gly Pro

100 105 110

Leu Pro Pro His Leu Ala Leu Lys Ala Glu Asn Asn Ser Glu Val Gly

115 120 125

Ala Ser Gly Tyr Gly Val Pro Gly Pro Thr Trp Asp Arg Gly Ala Asn

130 135 140

Leu Lys Asp Tyr Tyr Ser Arg Lys Glu Glu Gln Glu Val Gln Ala Thr

145 150 155 160

Leu Glu Ser Glu Glu Val Asp Leu Asn Ala Gly Leu His Gly Asn Trp

165 170 175

Thr Leu Glu Asn Ala Lys Ala Arg Leu Asn Gln Tyr Phe Gln Lys Glu

180 185 190

Lys Ile Gln Gly Glu Tyr Lys Tyr Thr Gln Val Gly Pro Asp His Asn

195 200 205

Arg Ser Phe Ile Ala Glu Met Thr Ile Tyr Ile Lys Gln Leu Gly Arg

210 215 220

Arg Ile Phe Ala Arg Glu His Gly Ser Asn Lys Lys Leu Ala Ala Gln

225 230 235 240

Ser Cys Ala Leu Ser Leu Val Arg Gln Leu Tyr His Leu Gly Val Val

245 250 255

Glu Ala Tyr Ser Gly Leu Thr Lys Lys Lys Glu Gly Glu Thr Val Glu

260 265 270

Pro Tyr Lys Val Asn Leu Ser Gln Asp Leu Glu His Gln Leu Gln Asn

275 280 285

Ile Ile Gln Glu Leu Asn Leu Glu Ile Leu Pro Pro Pro Glu Asp Pro

290 295 300

Ser Val Pro Val Ala Leu Asn Ile Gly Lys Leu Ala Gln Phe Glu Pro

305 310 315 320

Ser Gln Arg Gln Asn Gln Val Gly Val Val Pro Trp Ser Pro Pro Gln

325 330 335

Ser Asn Trp Asn Pro Trp Thr Ser Ser Asn Ile Asp Glu Gly Pro Leu

340 345 350

Ala Phe Ala Thr Pro Glu Gln Ile Ser Met Asp Leu Lys Asn Glu Leu

355 360 365

Met Tyr Gln Leu Glu Gln Asp His Asp Leu Gln Ala Ile Leu Gln Glu

370 375 380

Arg Glu Leu Leu Pro Val Lys Lys Phe Glu Ser Glu Ile Leu Glu Ala

385 390 395 400

Ile Ser Gln Asn Ser Val Val Ile Arg Gly Ala Thr Gly Cys Gly

405 410 415

Lys Thr Thr Gln Val Pro Gln Phe Ile Leu Asp Asp Phe Ile Gln Asn

420 425 430

Asp Arg Ala Ala Glu Cys Asn Ile Val Val Thr Gln Pro Arg Arg Ile

435 440 445

Ser Ala Val Ser Val Ala Glu Arg Val Ala Phe Glu Arg Gly Glu Glu

450 455 460

Pro Gly Lys Ser Cys Gly Tyr Ser Val Arg Phe Glu Ser Ile Leu Pro

465 470 475 480

Arg Pro His Ala Ser Ile Met Phe Cys Thr Val Gly Val Leu Leu Arg

485 490 495

Lys Leu Glu Ala Gly Ile Arg Gly Ile Ser His Val Ile Val Asp Glu

500 505 510

Ile His Glu Arg Asp Ile Asn Thr Asp Phe Leu Leu Val Val Leu Arg

515 520 525

Asp Val Val Gln Ala Tyr Pro Glu Val Arg Ile Val Leu Met Ser Ala

530 535 540

Thr Ile Asp Thr Ser Met Phe Cys Glu Tyr Phe Phe Asn Cys Pro Ile

545 550 555 560

Ile Glu Val Tyr Gly Arg Thr Tyr Pro Val Gln Glu Tyr Phe Leu Glu

565 570 575

Asp Cys Ile Gln Met Thr His Phe Val Pro Pro Pro Lys Asp Lys Lys

580 585 590

Lys Lys Asp Lys Asp Asp Asp Gly Gly Glu Asp Asp Asp Ala Asn Cys

595 600 605

Asn Leu Ile Cys Gly Asp Glu Tyr Gly Pro Glu Thr Arg Leu Ser Met

610 615 620

Ser Gln Leu Asn Glu Lys Glu Thr Pro Phe Glu Leu Ile Glu Ala Leu

625 630 635 640

Leu Lys Tyr Ile Glu Thr Leu Asn Val Pro Gly Ala Val Leu Val Phe

645 650 655

Leu Pro Gly Trp Asn Leu Ile Tyr Thr Met Gln Lys His Leu Glu Met

660 665 670

Asn Pro His Phe Gly Ser His Arg Tyr Gln Ile Leu Pro Leu His Ser

675 680 685

Gln Ile Pro Arg Glu Glu Gln Arg Lys Val Phe Asp Pro Val Pro Val

690 695 700

Gly Val Thr Lys Val Ile Leu Ser Thr Asn Ile Ala Glu Thr Ser Ile

705 710 715 720

Thr Ile Asn Asp Val Val Tyr Val Ile Asp Ser Cys Lys Gln Lys Val

725 730 735

Lys Leu Phe Thr Ala His Asn Asn Met Thr Asn Tyr Ala Thr Val Trp

740 745 750

Ala Ser Lys Thr Asn Leu Glu Gln Arg Lys Gly Arg Ala Gly Arg Val

755 760 765

Arg Pro Gly Phe Cys Phe His Leu Cys Ser Arg Ala Arg Phe Glu Arg

770 775 780

Leu Glu Thr His Met Thr Pro Glu Met Phe Arg Thr Pro Leu His Glu

785 790 795 800

Ile Ala Leu Ser Ile Lys Leu Leu Arg Leu Gly Gly Ile Gly Gln Phe

805 810 815

Leu Ala Lys Ala Ile Glu Pro Pro Pro Leu Asp Ala Val Ile Glu Ala

820 825 830

Glu His Thr Leu Arg Glu Leu Asp Ala Leu Asp Ala Asn Asp Glu Leu

835 840 845

Thr Pro Leu Gly Arg Ile Leu Ala Lys Leu Pro Ile Glu Pro Arg Phe

850 855 860

Gly Lys Met Met Ile Met Gly Cys Ile Phe Tyr Val Gly Asp Ala Ile

865 870 875 880

Cys Thr Ile Ala Ala Ala Thr Cys Phe Pro Glu Pro Phe Ile Asn Glu

885 890 895

Gly Lys Arg Leu Gly Tyr Ile His Arg Asn Phe Ala Gly Asn Arg Phe

900 905 910

Ser Asp His Val Ala Leu Leu Ser Val Phe Gln Ala Trp Asp Asp Ala

915 920 925

Arg Met Gly Gly Glu Glu Ala Glu Ile Arg Phe Cys Glu His Lys Arg

930 935 940

Leu Asn Met Ala Thr Leu Arg Met Thr Trp Glu Ala Lys Val Gln Leu

945 950 955 960

Lys Glu Ile Leu Ile Asn Ser Gly Phe Pro Glu Asp Cys Leu Leu Thr

965 970 975

Gln Val Phe Thr Asn Thr Gly Pro Asp Asn Asn Leu Asp Val Val Ile

980 985 990

Ser Leu Leu Ala Phe Gly Val Tyr Pro Asn Val Cys Tyr His Lys Glu

995 1000 1005

Lys Arg Lys Ile Leu Thr Thr Glu Gly Arg Asn Ala Leu Ile His

1010 1015 1020

Lys Ser Ser Val Asn Cys Pro Phe Ser Ser Gln Asp Met Lys Tyr

1025 1030 1035

Pro Ser Pro Phe Phe Val Phe Gly Glu Lys Ile Arg Thr Arg Ala

1040 1045 1050

Ile Ser Ala Lys Gly Met Thr Leu Val Thr Pro Leu Gln Leu Leu

1055 1060 1065

Leu Phe Ala Ser Lys Lys Val Gln Ser Asp Gly Gln Ile Val Leu

1070 1075 1080

Val Asp Asp Trp Ile Lys Leu Gln Ile Ser His Glu Ala Ala Ala

1085 1090 1095

Cys Ile Thr Gly Leu Arg Ala Ala Met Glu Ala Leu Val Val Glu

1100 1105 1110

Val Thr Lys Gln Pro Ala Ile Ile Ser Gln Leu Asp Pro Val Asn

1115 1120 1125

Glu Arg Met Leu Asn Met Ile Arg Gln Ile Ser Arg Pro Ser Ala

1130 1135 1140

Ala Gly Ile Asn Leu Met Ile Gly Ser Thr Arg Tyr Gly Asp Gly

1145 1150 1155

Pro Arg Pro Pro Lys Met Ala Arg Tyr Asp Asn Gly Ser Gly Tyr

1160 1165 1170

Arg Arg Gly Gly Ser Ser Tyr Ser Gly Gly Gly Tyr Gly Gly Gly

1175 1180 1185

Tyr Ser Ser Gly Gly Tyr Gly Ser Gly Gly Tyr Gly Gly Ser Ala

1190 1195 1200

Asn Ser Phe Arg Ala Gly Tyr Gly Ala Gly Val Gly Gly Gly Tyr

1205 1210 1215

Arg Gly Val Ser Arg Gly Gly Phe Arg Gly Asn Ser Gly Gly Asp

1220 1225 1230

Tyr Arg Gly Pro Ser Gly Gly Tyr Arg Gly Ser Gly Gly Phe Gln

1235 1240 1245

Arg Gly Gly Gly Arg Gly Ala Tyr Gly Thr Gly Tyr Phe Gly Gln

1250 1255 1260

Gly Arg Gly Gly Gly Gly Tyr

1265 1270

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Claims (34)

1. A method for preoperative risk stratification in a subject suffering from prostate cancer, comprising: - determining the expression profile of the gene for variant 7 of phosphodiesterase 4D (PDE4D7) in a biological sample obtained from the specified subject, - determining the expression profile of the gene for DExH-box helicase 9 (DHX9) in the same or different biological sample obtained from the specified subject, and - determining a preoperative predictive risk score for said subject based on said gene expression profile for PDE4D7 and said gene expression profile for DHX9, wherein said gene expression profile for PDE4D7 and gene expression profile for DHX9 are combined with a regression function that was derived from a population of prostate cancer subjects, and where the risk assessment indicates the risk of biochemical recurrence (BCR), clinical recurrence (progression to local and/or distant metastases) (CR), or specific death from prostate cancer. 2. The method of claim 1, further comprising: - suggestion of primary treatment for the specified subject based on the specified preoperative predictive risk assessment, wherein said primary treatment is selected from the group consisting of: (i) at least partial prostatectomy; (ii) active therapy selected from radiation therapy, hormonal therapy, chemotherapy, and combinations thereof; and (iii) active surveillance. 3. The method of claim 1, further comprising: - normalization of said gene expression profile for PDE4D7 and/or gene expression profile for DHX9 with respect to one or more reference genes selected from the group consisting of: Homo sapiens hypoxanthine phosphoribosyltransferase 1 (HPRT1), tubulin alpha-1b (TUBA1B), member Homo sapiens pumilio RNA-binding family (PUM1) and Homo sapiens TATA box-binding protein (TBP), wherein said preoperative predictive risk score is determined based on said normalized gene(s) expression profile(s). 4. The method of claim 3, wherein said one or more reference genes contain at least two, or at least three, or all of HPRT1, TUBA1B, PUM1, and TBP. 5. The method according to claim 4, characterized in that said determination of a gene expression profile for PDE4D7 and/or a gene expression profile for DHX9 comprises performing a quantitative real-time polymerase chain reaction (RT-qPCR) on RNA isolated from biological (their ) sample(s), wherein a Cq value is determined for PDE4D7 and/or DHX9 and for each of one or more reference genes, wherein said preoperative risk assessment definition includes normalizing said Cq value for PDE4D7 and/or DHX9 using said Cq value for each of the one or more reference genes and calculating said preoperative risk score based on said normalized Cq value(s). 6. Diagnostic kit for determining preoperative prognostic risk assessment in a patient suffering from prostate cancer, containing: at least one primer and/or probe for profiling the expression of a gene for variant 7 of phosphodiesterase 4D (PDE4D7) in a biological sample obtained from a subject suffering from prostate cancer; at least one primer and/or probe for profiling gene expression for DExH box helicase 9 (DHX9) in the same or a different biological sample obtained from said subject; And optionally at least one primer and/or probe for gene expression profiling for one or more reference genes selected from the group consisting of: Homo sapiens hypoxanthine phosphoribosyltransferase 1 (HPRT1), tubulin alpha-1b (TUBA1B), RNA-binding member Homo sapiens pumilio (PUM1) family and Homo sapiens TATA box-binding protein (TBP); said diagnostic kit further comprising instructions for calculating a preoperative predictive risk score based on said gene expression profile for PDE4D7 and gene expression profile for DHX9, said instructions being stored in a computer program product which, when executed by a computer, performs a method comprising: - determining said preoperative predictive risk score for said subject based on said gene expression profile for PDE4D7 and gene expression profile for DHX9, wherein optionally said method comprises: - normalizing said gene expression profile for PDE4D7 and/or gene expression profile for DHX9 with respect to one or more reference genes, wherein said preoperative predictive risk score is determined based on said normalized gene(s) expression profile(s). 7. Diagnostic kit according to claim 6, characterized in that said instructions for calculating said preoperative predictive risk score contain instructions for combining said gene expression profile for PDE4D7 and gene expression profile for DHX9 with a regression function that was obtained based on a population of subjects, suffering from prostate cancer. 8. Use of a diagnostic kit according to claim 6 or 7 in a method for preoperative risk stratification in a subject suffering from prostate cancer. 9. Use of a gene expression profile for phosphodiesterase 4D variant 7 (PDE4D7) and a gene expression profile for DExH-box helicase 9 (DHX9) in preoperative risk stratification in a subject suffering from prostate cancer, including: - determining said gene expression profile for PDE4D7 in a biological sample obtained from said subject, - determining said gene expression profile for DHX9 in the same or different biological sample obtained from said subject, and - determining a preoperative predictive risk score for said subject based on said gene expression profile for PDE4D7 and gene expression profile for DHX9. 10. Use according to claim 9, characterized in that said gene expression profile for PDE4D7 and gene expression profile for DHX9 are combined with a regression function that has been derived from a population of prostate cancer subjects. 11. A computer-readable medium on which a control program is recorded, containing instructions that, when the specified program is executed by a computer, leads to the implementation by the computer of a method, including: - determining a preoperative predictive risk score for a subject suffering from prostate cancer based on the gene expression profile for phosphodiesterase 4D variant 7 (PDE4D7) and the gene expression profile for DExH-box helicase 9 (DHX9), wherein said gene expression profile for PDE4D7 was determined in a biological sample obtained from said subject, and said gene expression profile for DHX9 was determined in the same or another biological sample obtained from said subject. 12. A machine-readable carrier according to claim 11, characterized in that said gene expression profile for PDE4D7 and gene expression profile for DHX9 are combined with a regression function that was derived from a population of subjects with prostate cancer.

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