CN109423515B - Gene markers for liver cancer detection and application thereof - Google Patents

Abstract

The invention discloses a group of genetic markers for liver cancer detection and application thereof, and is particularly suitable for detection of early liver cancer and/or detection of AFP negative liver cancer. A group of liver cancer gene markers comprises an EP400 gene, an MAPK1IP1L gene, a NUFIP2 gene, a PHC3 gene, an RPS6KB1 gene and an STX7 gene. The invention has the advantages that: the gene marker of the invention is used for the screening diagnosis of liver cancer, has high sensitivity and strong specificity, is convenient to detect because the peripheral blood which is most easily collected clinically is taken as a detection sample, is suitable for the early diagnosis of liver cancer and the detection of AFP negative liver cancer patients, and has wide application prospect.

Description

Gene markers for liver cancer detection and application thereof

Technical Field

The invention relates to a group of gene markers for liver cancer detection and application thereof, in particular to the detection of early liver cancer and/or the detection of AFP negative liver cancer, and the application field is biological science, biochemistry, biotechnology, medicine and medical technology.

Background

Liver cancer is one of malignant tumors that seriously affect the health of our country. According to the latest published annual registration report for tumors 2014 published by national tumor registration center, the morbidity accounts for 4 th of all malignant tumors, and the mortality accounts for 2 nd of all cancer deaths, and is only second to lung cancer. Epidemiological statistics show that the high-incidence age of liver cancer of Chinese people is advanced from 40-60 years to 30-60 years before. The liver cancer is in the trend of youthful onset, and seriously damages the population quality, social stability and economic construction of each region in China.

The expert consensus of 'standardized diagnosis and treatment of primary liver cancer' in 2009 indicates that: the primary liver cancer is a high-incidence tumor in China, and has more difficulties in treatment; therefore, early detection and early diagnosis must be considered. The treatment means and prognosis of liver cancer are closely related to the course of disease of patients: the patients with earlier disease course can adopt treatment methods such as surgical excision, liver transplantation and the like, and the 5-year survival rate can reach 50-70%; for patients with a later course of disease, the measures such as hepatic artery chemotherapy and targeted therapy can be adopted, and the 3-year survival rate is only 10-40%. Early diagnosis of liver cancer is the best measure for improving surgical resection rate, reducing mortality rate, prolonging survival time and improving survival quality.

The liver cancer is screened internationally by mainly applying a diagnosis technology combining Alpha Fetoprotein (AFP), ultrasound and other imaging, but the ultrasound examination depends on the experience and equipment of an operator, while other imaging procedures require high cost, and the liver cancer is generated in an early stage and cannot be examined by the imaging; the AFP sensitivity is low, and the AFP is difficult to be used for early diagnosis and is easy to cause missed diagnosis or misdiagnosis of diseases. Early diagnosis of treatment delays, so that more than two-thirds of liver cancer patients are treated in the late stage. At present, 38.76 million new liver cancer patients are added in China every year, wherein the number of AFP negative liver cancer patients is about 11.6 million, and the liver cancer patients belong to a high-risk liver cancer patient group which is easy to delay the best treatment opportunity due to missed diagnosis and misdiagnosis.

AFP negative liver cancer patients account for about 30 percent of the total number of liver cancer patients, small liver cancer (tumor body is less than or equal to 3cm) is particularly common, and is mostly low-differentiation liver cancer, the prognosis is poor, and the clinical cure rate can be improved by early diagnosis and early surgical excision. The AFP level of serum of an AFP negative liver cancer patient is normal (< 20 mu g/L), the clinical symptoms are light and lack of specificity, the current clinical diagnosis mostly depends on the imaging and pathological examination, particularly, the AFP negative liver cancer with the tumor less than 3cm lacks valuable and practical diagnostic markers, and the diagnosis is easily misdiagnosed as a liver benign disease, so the treatment is delayed. How to diagnose AFP negative liver cancer is a considerable problem. The combined detection of other tumor markers is a preferable method for diagnosing liver cancer, especially AFP negative liver cancer.

At present, tumor biomarkers are generally obtained from tumor tissue specimens or peripheral blood. Blood is a connective tissue that circulates in a broad sense, contacts all organs of the human body, contains indexes reflecting changes of the body, and participates in disease processes in pathological states; compared with the sampling of tumor tissue samples, the acquisition of the markers in the blood circulation is non-invasive, has the advantages of repeatable sampling and dynamic monitoring, and is an ideal tissue for replacing the biopsy of pathological parts and carrying out non-invasive molecular detection. The study of novel tumor markers at the level of transcriptomes including messenger rna (mrna), micro-rna (mirna) and nucleic acid mutations including circulating tumor dna (ctdna) is gaining increasing attention. Compared with the traditional serum protein molecular marker, the nucleic acid molecular marker has obvious advantages in stability, detection sensitivity and detection flux, can better meet the requirement of multi-marker combined detection to improve the diagnosis rate, and is a development trend of tumor marker research and development and detection.

Research shows that the peripheral blood transcriptome can accurately display the expression characteristics of disease-specific genes and can be applied to early diagnosis and prognosis judgment of diseases. Based on the specific gene expression profile of peripheral blood and combined with the fluorescent quantitative PCR technology, England scientists detect the expression condition of an 8-gene characteristic profile in kidney cancer as an early diagnosis index of renal cell carcinoma and obviously related to the overall survival period and the progression-free survival period of patients. Compared with miRNA, the research on the expression profile of the tumor-related mRNA is more thorough, the detection method is more mature, and the result is more accurate. Gene diagnosis products for early warning of risk of intestinal cancer based on mRNA level in the united states have been developed and marketed.

Disclosure of Invention

The first technical problem to be solved by the invention is to provide a group of effective novel gene markers for liver cancer detection based on peripheral blood gene expression characteristic pedigrees.

Another technical problem to be solved by the present invention is to provide the use of the above gene marker.

In order to achieve the purpose, the invention adopts the following technical scheme:

in the first aspect of the invention, a group of liver cancer gene markers is provided, which consists of an EP400 gene, a MAPK1IP1L gene, a NUFIP2 gene, a PHC3 gene, an RPS6KB1 gene and an STX7 gene. The research of the invention discovers for the first time that the genes are differentially expressed in peripheral blood of liver cancer patients (especially AFP negative liver cancer patients).

The EP400 gene (Homo sapiens E1A binding protein p400, human E1A binding protein p400) is a polynucleotide sequence shown in SEQ ID NO:1 of the sequence Listing (NCBI accession No. NM _015409), the MAPK1IP1L gene (Homo sapiens mitogen-activated protein kinase 1interacting protein 1like human mitogen-activated protein kinase 1interacting protein 1) is a polynucleotide sequence shown in SEQ ID NO:2 of the sequence Listing (NCBI accession No. NM _144578), the NUFIP2 gene (Homo sapiens NUFIP2, FMR1interacting protein 2, human FMR1interacting protein 2) is a polynucleotide sequence shown in SEQ ID NO:3 (NCBI accession No. 020772), the PHC3 gene (Homo sapiens accession No. E3, human homolog 3 homolog accession No. 2) is a polynucleotide sequence shown in SEQ ID NO: 465 homolog of SEQ ID NO: PIB 3 homolog, the human homolog of the sequence No. 2 (NCBI accession No. 7) is a polynucleotide sequence shown in SEQ ID NO: 465 homologous polynucleotide sequence shown in SEQ ID NO:5 (NCBI accession No. 4) of the sequence No. 4 NCBI number NM _003161), STX7 gene (Homo sapiens syntaxin 7, human synapsin 7) is a polynucleotide sequence (NCBI number NM _003569) shown in a sequence table SEQ ID NO. 6.

The method is based on the quantitative analysis of the human whole-gene expression profile, compares the gene expression difference in the peripheral blood samples of the liver cancer patient and the normal human by quantitatively analyzing the gene expression condition of the total RNA of the peripheral blood, and screens out 6 gene signals which are differentially expressed (the gene expression is up-regulated or down-regulated) in the peripheral blood of the liver cancer patient by a Support Vector Machine (Support Vector Machine) method. Then, the relative expression of the 6 gene markers in the peripheral blood of the detected person can be quantitatively detected by adopting fluorescent quantitative RT-PCR, gene expression profiling chip or RNA sequencing technology, so as to judge the probability of the detected person suffering from liver cancer.

In a second aspect of the invention, the application of the reagent for detecting the liver cancer gene marker in the preparation of a product for detecting liver cancer is provided.

The products for detecting liver cancer include, but are not limited to, real-time quantitative PCR kit, RNA sequencing kit or gene chip.

The reagent for detecting the liver cancer gene marker is a primer and/or a probe for detecting the liver cancer marker.

The primer is a primer for specifically amplifying SEQ ID NO:1 to SEQ ID NO:6 gene sequence. Preferably, the primer has a sequence shown in a sequence table SEQ ID NO. 7-SEQ ID NO. 18.

The probe is a probe which can be matched with SEQ ID NO:1 to SEQ ID NO: 6. Preferably, the probe has a sequence shown in a sequence table SEQ ID NO. 19-SEQ ID NO. 24.

In a third aspect of the present invention, a kit for detecting liver cancer is provided, which comprises a nucleic acid sequence specifically directed to SEQ ID NO:1 to SEQ ID NO: 6.

Preferably, the sequence of the primer is SEQ ID NO:7 to SEQ ID NO:18 is shown in the figure; the sequence of the probe is SEQ ID NO:19 to SEQ ID NO: as shown at 24.

The kit also comprises a primer and/or a probe aiming at an internal reference gene GAPDH (NM-002046.5, Homo sapiens glyceraldehyde-3-phosphate dehydrogenase). The forward primer sequence was ACCATGAGAAGTATGACAACAGCC (SEQ ID NO: 25 of the sequence Listing), the reverse primer sequence was CACGATACCAAAGTTGTCATGGA (SEQ ID NO: 26 of the sequence Listing), and the probe sequence was TCAGCAATGCCTCCTGCACCACC (SEQ ID NO: 27 of the sequence Listing). GAPDH is an enzyme in glycolysis reactions, widely distributed in cells in various tissues. The enzyme gene is housekeeping (house keeping) gene, is expressed at high level in almost all tissues, and is a standardized internal reference of the common fluorescent quantitative PCR experiment operation.

The probe sequence has a fluorescent label, the 5 'end fluorescent label is one of FAM, HEX, TET, TAMRA, Cy5, Cy3, VIC, R0X and JOE, and the 3' end fluorescent label is one of TAMRA, BHQ, MGB, DABCYL and Elipse.

By using the kit, the kit can detect the nucleotide sequence of SEQ ID NO:1 to SEQ ID NO:6, and then judging the probability of the liver cancer of the examined person according to the information of the up-regulation or down-regulation of the gene expression, thereby realizing the screening and diagnosis of the liver cancer.

All kits described herein can contain suitable packaging and instructions for use in the methods disclosed herein. The kit may further comprise suitable buffers and a polymerase, such as a thermostable polymerase, e.g., Taq polymerase. Such kits also comprise control primers and/or probes.

According to the inventionThe detection kit is a nucleic acid detection kit and comprises a peripheral blood total RNA extraction reagent and a PCR reaction system reagent, wherein the PCR reaction system comprises dNTP and MgCl₂Taq DNA polymerase, PCR reaction buffer, primers and probes.

In a fourth aspect of the invention, a method for detecting liver cancer is provided, and determining the presence and/or level of the panel of liver cancer gene biomarkers can comprise determining the presence and/or level of a polynucleotide (e.g., a biomarker gene transcript) encoding the biomarker. The method comprises the following steps:

(1) determining the presence and/or level of a hepatoma gene marker in a sample of a subject; and

(2) comparing the presence and/or level of the liver cancer gene marker to a control, wherein a presence and/or level in the sample that is different from the control is indicative of liver cancer.

The sample comprises blood, plasma, serum, urine, platelets, megakaryocytes (megakaryocytes), or excreta.

In a fifth aspect of the present invention, there is provided a method for treating liver cancer, the method comprising:

(1) diagnosing or detecting liver cancer according to the methods of the invention; and

(2) administering or recommending a therapeutic agent for treating liver cancer.

The invention uses the peripheral blood gene marker for liver cancer screening, the marker is used for screening and diagnosing liver cancer, has high sensitivity and specificity, and is simple and convenient in detection process due to the adoption of peripheral blood which is most easily collected clinically, thus being suitable for diagnosing early liver cancer, in particular AFP negative liver cancer. The screening means of combining Alpha Fetoprotein (AFP), ultrasound and other imaging which are mainly applied to liver cancer diagnosis at present is improved, so that the omission of AFP negative liver cancer is reduced, the dependence on experience and equipment of operators is reduced, and a novel diagnosis method is provided for early diagnosis of liver cancer. The research and development of peripheral blood liver cancer molecular diagnosis products can meet the development direction and social requirements of clinical diagnosis and treatment of liver cancer, is favorable for further promoting the popularization of liver cancer screening, improves the detection rate of liver cancer patients and the success rate of clinical treatment, and saves social capital and corresponding medical resources.

The invention has the advantages that: the gene marker of the invention is used for the screening diagnosis of liver cancer, has high sensitivity and strong specificity, is convenient to detect because the peripheral blood which is most easily collected clinically is taken as a detection sample, is suitable for the early diagnosis of liver cancer and the detection of AFP negative liver cancer patients, and has wide application prospect.

The invention will be further illustrated with reference to the following specific examples. These examples are intended to illustrate the invention and are not intended to limit the scope of the invention. The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out according to conventional conditions or according to conditions recommended by the manufacturers. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In addition, any methods and materials similar or equivalent to those described herein can be used in the methods of the present invention. The preferred embodiments and materials described herein are intended to be exemplary only.

Drawings

FIG. 1 shows the logarithmic fluorescence intensity values after normalization processing for 100 chips in example 1.

FIG. 2 is a diagram showing the accuracy, sensitivity and specificity of the liver cancer and normal control samples randomly sampled by using the selected liver cancer gene markers in example 3 of the present invention.

Detailed Description

The term "training set" or "control set" refers to a set of samples used to establish a correlation between two variables. For example, a training set is a set of patient samples used to establish correlations between gene expression and the condition of a patient. In the context of the present invention, a training set is a set of samples from liver cancer patients, hepatitis patients, normal persons for which pathologically relevant information including clinical diagnosis, tumor size, serum AFP expression, etc. can be obtained. The training set was used to establish correlations between expression profiles and patient disease classification (including serum AFP expression).

The term "test set" refers to a set of samples used to validate correlations established using a training set. For example, a test set is a set of samples from patients whose gene expression and patient condition are known. This set is used to verify whether the correlations determined using the training set will correctly predict the patient's condition. In the context of the present invention, a test set is a set of samples from liver cancer patients and normal persons for which a patient disease classification (including serum AFP expression) is available. The test set is used to measure gene expression in the profile and test whether the actual patient disease classification matches the prediction made by measuring gene expression.

Example 1 screening of markers for liver cancer characteristic genes

Materials and methods

(A) Material

The inventor entrusts Shenzhen hospital of Beijing university to collect a large number of peripheral blood samples of normal people, liver cancer patients and liver benign disease patients in 11 months to 2013 months in 2012 (the samples for research are collected, sampled, subpackaged and stored under uniform conditions in the same period), and through sorting sample data, the inventor selects 81 patient plasma (hepatitis B-liver cancer (including primary liver cancer and intrahepatic bile duct cancer)) and 19 plasma of normal people randomly collected in the same period with the hospital and an experimental group, and avoids taking plasma samples of people with family history of liver cancer as much as possible to detect mRNA.

Hepatocellular carcinoma sample inclusion met both of the following criteria: firstly, a pathological diagnosis standard: biopsy or surgical resection of tissue specimens of liver space occupying lesions or extrahepatic metastases is diagnosed as HCC by histopathological and/or cytological examination, which is the gold standard. ② clinical diagnosis standard: clinical diagnosis of HCC can be established when two items (1) + (2) a or three items (1) + (2) b + (3) are required to be simultaneously satisfied. (1) Evidence of cirrhosis and HBV and/or HCV infection (HBV and/or HCV antigen positive); (2) typical HCC imaging characteristics are that contemporaneous multi-row CT scans and/or dynamic contrast-enhanced MRI examinations show rapid heterogeneous vascular reinforcement (hepatic hypervariability) of liver occupancy at the arterial phase, and rapid elution (venous or delayed phase wash out) at the venous phase. a, if the occupying diameter of the liver is 2cm, one of CT and MRI two imaging examinations shows that the occupying area of the liver has the characteristics of the liver cancer, and HCC can be diagnosed; b, if the occupied diameter of the liver is 1-2 cm, CT and MRI two-image examination is needed to show that the occupied area of the liver has the characteristics of the liver cancer, so that HCC can be diagnosed, and the specificity of diagnosis is enhanced. (3) Serum AFP 400 μ g/L lasts for 1 month or 200 μ g/L lasts for 2 months, and can eliminate AFP increase caused by other reasons, including pregnancy, germline embryonal tumor, active liver disease and secondary liver cancer.

After the discussion of the ethical committee of the hospital passed and the patients signed the informed consent, the patient peripheral blood samples were collected.

(II) method

1. Extracting and purifying total RNA in peripheral blood by using PAXgeneblood RNA Kit of QIAGEN company, and identifying the integrity and yield of the extracted total RNA fragment by using an Agilent BioAnalyzer model 2100 micro electrophoresis analyzer;

2. detecting the Gene Expression profile of peripheral blood mRNA of 54 patients (comprising 25 AFP negative liver cancers and 29 AFP positive liver cancers), 9 patients (AFP negative) diagnosed with intrahepatic bile duct cancer, 18 patients (comprising 12 AFP negative hepatitis and 6 AFP positive hepatitis) diagnosed with hepatitis and 19 normal persons by adopting an Affymetrix Gene Expression Using Affymetrix U133Plus 2.0 human whole-Gene Expression profile chip (Affymetrix, Inc. Santa Clara, Calif.);

3. chip data preprocessing: first, the background of the probe intensity was corrected by the MAS5 method, and the normalization process was performed on 100 chips. FIG. 1 shows the logarithmic fluorescence intensity values after normalization processing for 100 chips.

4. Removing probe set (Probe set) signals with excessively high and excessively low expression signals in the total RNA, and selecting probe set signals with moderate expression (between the fluorescence signal value of 100-10000) in all 100 samples in the training set for subsequent analysis;

5. selecting differentially expressed genes of liver cancer, hepatitis and normal people by adopting a T test method, and screening a probe set signal under the condition that the fold change FC is more than 1.2 times and the significance level P is less than 0.05 to obtain 248 groups of probe sets meeting the requirements;

6. screening liver cancer characteristic genes from the 248 probe sets by using a Support Vector Machine method (Support Vector Machine), constructing a binary mathematical model according to an SFFS (sequential Forward flow selection) screening strategy, randomly sampling training set samples by using a LOOCV (leave-one-out cross identification) method, classifying and predicting the sample groups generated by sampling, selecting 6 genes with the highest prediction accuracy (accuracy) for liver cancer as liver cancer candidate characteristic genes, and establishing a screening diagnosis model of liver cancer and normal persons.

Secondly, the result is:

by analyzing and comparing the gene expression profiles of different types of samples and utilizing correlation analysis, the characteristic gene expression profile which can most distinguish AFP negative liver cancer from a control sample is screened out. A group of liver cancer characteristic gene markers are obtained, are an AFP negative liver cancer characteristic expression gene catalogue (gene panel), and consist of an EP400 gene, an MAPK1IP1L gene, a NUFIP2 gene, a PHC3 gene, an RPS6KB1 gene and an STX7 gene.

The AFP negative liver cancer characteristic marker obtained by the invention consists of 6 genes shown in Table 1.

TABLE 1 liver cancer characteristic Gene marker sequences

Serial number	Name of liver cancer characteristic gene	Sequence Listing numbering
			1	EP400 gene	SEQ ID NO：1
2	MAPK1IP1L gene	SEQ ID NO：2
			3	NUFIP2 gene	SEQ ID NO：3
4	PHC3 gene	SEQ ID NO：4
			5	RPS6KB1 gene	SEQ ID NO：5
6	STX7 gene	SEQ ID NO：6

Example 2 fluorescent quantitative RT-PCR kit for detecting liver cancer

The kit comprises the following components:

1. specific primers for detecting 6 liver cancer characteristic gene markers (shown in table 1):

reagent 1: EP400 gene forward primer: 5'GGATCTTGTCAGTGACGTTGT 3' (SEQ ID NO: 7)

Reagent 2: EP400 gene reverse primer: 5'GTAGCGATTCCGGCACTGT 3' (SEQ ID NO: 8)

Reagent 3: MAPK1IP1L gene forward primer: 5'ACCTGAGGCAGCTCTTTTGC 3' (SEQ ID NO: 9)

Reagent 4: MAPK1IP1L gene reverse primer: 5'GGATGTGTGTGCTCCTTCTAAAGAA3' (SEQ ID NO: 10)

And (5) reagent: NUFIP2 gene forward primer: 5'TTTCTCTCAAAGGACTACGAGAT 3' (SEQ ID NO: 11)

Reagent 6: NUFIP2 gene reverse primer: 5'AGCAGCCCTCAGGTCAAA 3' (SEQ ID NO: 12)

And (7) reagent: PHC3 gene forward primer: 5'CAGTTCTACCAGCGGCAGTA 3' (SEQ ID NO: 13)

Reagent 8: PHC3 gene reverse primer: 5'GGTAGCGGGTGTGAAAATCA 3' (SEQ ID NO: 14)

Reagent 9: RPS6KB1 gene forward primer: 5'CCGAACTCTGGGCCATACA 3' (SEQ ID NO: 15)

Reagent 10: RPS6KB1 gene reverse primer: 5'TTGCAGGATGCTCACACATCTC 3' (SEQ ID NO: 16)

Reagent 11: STX7 gene forward primer: 5'CTCTTTTGTGAGTGAGTGATTGGAA 3' (SEQ ID NO: 17)

Reagent 12: STX7 gene reverse primer: 5'CCCTGCATTGTCCATTCTGTT 3' (SEQ ID NO: 18)

2. Detecting fluorescent quantitative PCR probe sequences of 6 liver cancer characteristic gene markers (shown in table 1), wherein the probe markers of 6 target genes are FAM markers at the 5 'end and MGB markers at the 3' end:

reagent 13: EP400 gene probe sequence: 5'AACTCCTGTAGCCGAATCTACCGCTC 3' (SEQ ID NO: 19)

Reagent 14: MAPK1IP1L gene probe sequence: 5'TAGCCACCCCCACCCCACTTGC 3' (SEQ ID NO: 20)

Reagent 15: NUFIP2 gene probe sequence: 5'TCAAAATCCTCTGGCCTCTCCTACGAAC3' (SEQ ID NO: 21)

Reagent 16: PHC3 gene probe sequence: 5'CCCCTACCCTAACGGCAAGCCA3' (SEQ ID NO: 22)

Reagent 17: RPS6KB1 gene probe sequence: 5'CAAACGGCCAGAGCACCTGCGT3' (SEQ ID NO: 23)

Reagent 18: STX7 gene probe sequence: 5'ACTCCTGTTGCCAGAATCAGACTGCCCTA 3' (SEQ ID NO: 24)

3. Control Gene (SEQ ID NO: 28) detection reagent

Reagent 19: GAPDH gene forward primer: ACCATGAGAAGTATGACAACAGCC (SEQ ID NO: 25)

Reagent 20: GAPDH gene reverse primer: CACGATACCAAAGTTGTCATGGA (SEQ ID NO: 26)

Reagent 21: GAPDH gene probe sequence: TCAGCAATGCCTCCTGCACCACC (SEQ ID NO: 27) GAPDH was labeled 5 'with VIC and 3' with MGB.

Second, use method

The reagents are core reagents for forming the liver cancer detection kit, and each reagent is independently packaged. On the basis, other reagents such as total RNA extraction reagent, PCR reaction solution, Taq enzyme system, packaging material and the like can be added, and are all conventional reagents in the field.

The primers and probes were used at a concentration of 10. mu.M, and the kit was used as described in example 3.

Example 3 screening diagnosis of Normal human by Using the screened marker for liver cancer characteristic Gene

A method step

1. Collecting peripheral blood samples of the sample to be detected: peripheral blood samples of patients were collected using BD PAXgeneRNA blood collection tubes from QIAGEN corporation. The sample sources were the same as in example 1.

2. Extracting and purifying total RNA in a peripheral blood sample of a sample to be detected: extracting and purifying total RNA in peripheral blood by using PAXgeneblood RNA Kit of QIAGEN company, and identifying the integrity and yield of the extracted total RNA fragment by using an Agilent BioAnalyzer model 2100 micro electrophoresis analyzer;

3. reverse transcription reaction: performing Reverse Transcription reaction by using a High-Capacity cDNA Reverse Transcription kit of Life Techonlgy company, using total RNA as a template and Random primers as Reverse Transcription primers to synthesize cDNA;

table 2: reaction system

Reaction components	Volume (μ l)
		10×RT Buffer	2
25×dNTP mix	0.8
		10×RT Random Primers	2
MultiScribeTM Reverse Transcriptase	1
		RNase Inhibitor	1
mRNA	Volume containing 1. mu.g of mRNA
		Nuclease-free H2O	Make up to a total volume of 20. mu.l

Table 3: reaction conditions

Is provided with	Step 1	Step 2	Step 3	Step 4
					Temperature of	25℃	37℃	85℃	4℃
Time
		10 minutes	120 minutes	5 minutes	∞

4. Fluorescent quantitative RT-PCR detection:

based on the related sequences of 6 liver cancer characteristic genes EP400 gene, MAPK1IP1L gene, NUFIP2 gene, PHC3 gene, RPS6KB1 gene, STX7 gene (SEQ ID NO: 1-SEQ ID NO: 6) and internal reference gene GAPDH (SEQ ID NO: 28), the primers of example 2, SEQ ID NO:7 to SEQ ID NO: 18. SEQ ID NO: 25. SEQ ID NO: 26 as a primer, and a specific probe SEQ ID NO:19 to SEQ ID NO: 24. SEQ ID NO: 27 as a probe, and cDNA obtained by reverse transcription is used as an amplification template to carry out real-time fluorescent quantitative RT-PCR reaction to obtain the mRNA relative content of the 6 gene markers in peripheral blood samples. Table 4 shows the composition of the PCR reaction premix (Master Mix), wherein the forward primer, reverse primer and probe refer to the forward primer, reverse primer and probe of each individual gene among EP400 gene, MAPK1IP1L gene, NUFIP2 gene, PHC3 gene, RPS6KB1 gene and STX7 gene. The system for fluorescent quantitative PCR detection is that a single target gene and a primer probe of internal reference GAPDH are mixed to form a double PCR system, and each target gene is detected once.

Table 4: reaction System exemplified by Taqman Probe method

Component (A)	×1(μl)
		PCR reaction premix(2×)	10
Forward primer (10. mu.M)	1.8
		Reverse primer (10. mu.M)	1.8
Probe (10 μ M)	0.5
		GAPDH forward primer (10. mu.M)	1.8
GAPDH reverse primer (10. mu.M)	1.8
		GAPDH probe (10. mu.M)	0.5
Blood sample cDNA (20 ng/. mu.l)	1
		H2O	0.8

Table 5: reaction conditions

5. Diagnosis of donor results of samples to be tested: and (3) judging whether the examined person suffers from the liver cancer or not according to the mRNA relative content of 6 gene markers in the peripheral blood sample in the fluorescent quantitative RT-PCR.

Secondly, analyzing the results

And storing the detection data file after the reaction is finished. The Start value, the End value and the Threshold value of Baseline are adjusted according to the analyzed image (the user can adjust the values according to the actual situation, the Start value can be I-10, the stop value can be 5-20, the Threshold value can be selected in the range of 5K-50K), so that the Standard curve chart under the 'Standard curve' window is optimized, namely the R2 value (correlation value) is 0.97. And finally, under a Report window, a recording instrument automatically analyzes the calculated copy number of the unknown specimen of the E2A-PBX1 (E2A-PBX1-Qty), and the result is derived. And (4) resetting the 4 positive quantitative reference products of the TBP according to the corresponding sequence, and deriving the copy number (TBP-Qty) of the TBP unknown specimen by the same steps.

Thirdly, judging the result

If the growth curve does not present an S-shaped curve or the Ct value is blank, the detected gene expression level is judged to be less than the detection limit.

If the growth curve is S-shaped and the Ct value of the sample<37, calculating the expression quantity (delta Ct) of each target gene relative to the internal reference in the sample, wherein the delta Ct is Ct_{(target Gene)}-Ct_{(internal reference gene)}And then calculating the logistic regression value of the sample by adopting the following formula: logit 4.2715 Δ Ct_(MAPK1IP1L)+1.1987*ΔCt_(RPS6KB1)-1.2091*ΔCt_(NUFIP2)+0.2748*ΔCt_(PHC3)+1.9441*ΔCt_(STX7)+0.4728*ΔCt_(EP400)+18.1326。

When the Logit value of the sample to be detected is greater than 2.30873, judging the property of the sample to be liver cancer; when the Logit value of the sample to be detected is < -1.67951, judging the property of the sample to be normal; when the Logit value of the sample to be detected is not less than 1.67951 and not more than 2.30873, the method cannot be used for judging the property of the sample.

The relative expression quantity of 6 liver cancer gene markers in 53 liver cancer confirmed patients (comprising 24 AFP negative liver cancers and 29 AFP positive liver cancers) and 19 normal persons peripheral blood is detected by fluorescent quantitative RT-PCR, the samples (comprising Training set and Test set) are classified and predicted by using a liver cancer screening diagnosis model, the prediction result is compared with the clinical diagnosis detection result of the liver cancer, the Sensitivity (Sensitivity), Specificity (Specificity) and Accuracy (Accuracy) of the 6 liver cancer gene markers of the invention on the screening diagnosis of the liver cancer (comprising AFP negative liver cancers) are obtained, and the specific detection result is shown in a figure 2 and a table 6.

Table 6: sensitivity, specificity and accuracy for discriminating and detecting liver cancer and normal person

The above-mentioned embodiments only express the embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention.

Sequence listing

<110> Shanghai biochip Co., Ltd

<120> a group of gene markers for liver cancer detection and application thereof

<130>

<160> 28

<170> PatentIn version 3.5

<210> 1

<211> 12317

<212> DNA

<213> genus, species, Homo sapiens (human)

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gcggcttcca tcctcccgcc ctcctgacgc ggccggagcg cagccctgag gcccagggag 120

aacgacacat tggatacaga agggaggtga tcatgcacca tggcactggc ccccagaacg 180

tccagcatca gctgcagagg tccagggcct gccctggcag cgagggtgag gagcagccgg 240

cccaccccaa cccacccccg tcccccgcag ctcccttcgc tccctcagca agcccgtcgg 300

caccccagtc tcccagttat caaatacagc agctgatgaa taggagccct gcaaccgggc 360

agaacgtgaa catcaccctg cagagcgtgg gccctgtcgt cgggggaaac cagcagatca 420

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aggtccagac ccagagtccc acgcagccca gtccggggcc ggggcaggcc ttgcagaatg 600

tgcgtgcagg tgcccctggc cctgggctgg gcctctgcag cagcagccct acagggggct 660

tcgtggatgc cagcgtgctg gtgaggcaga tcagcttgag cccctccagt ggtggacact 720

ttgtgtttca ggatgggtca gggctcaccc agatcgccca gggagcccag gttcagctcc 780

agcacccggg tacgcccatc acagtccgag agcggagacc ctcccagccc cacacacagt 840

cagggggcac catccaccac ctgggacccc agagccctgc agccgcgggt ggggccggcc 900

tgcagcccct ggccagccca agccacatca ccacggctaa cttgccaccg cagatcagca 960

gcatcatcca gggccagctg gttcagcagc agcaggtgct gcaggggccg ccgctgcccc 1020

ggcccctggg cttcgagagg acacccggcg tgctgctccc cggggctggg ggcgcagcgg 1080

ggtttgggat gacgtcccca cccccgccca ccagcccttc caggactgcc gtgcccccag 1140

gcctttccag cctcccactc acgtctgtgg ggaacacggg aatgaagaag gttcccaaga 1200

agttagagga gattccccca gcctctccgg agatggcaca gatgaggaag cagtgcctgg 1260

actatcatta ccaggagatg caggctctga aggaggtctt caaggagtat ttgattgaac 1320

tgtttttctt gcaacacttt caagggaaca tgatggattt cttagctttc aagaagaaac 1380

attatgcccc attacaagca tatcttaggc agaatgattt ggacattgaa gaagaggagg 1440

aggaggagga agaggaggaa gaaaaatctg aggttatcaa tgacgagcag caagccctcg 1500

cagggagcct ggtagcaggg gccggaagca cagtagagac ggacctgttt aagaggcagc 1560

aggcgatgcc ctccacaggt atggcagagc agtctaagag gcctcgcctt gaagtgggtc 1620

accaaggggt agttttccag cacccagggg cggacgcagg cgttcctctc cagcaactaa 1680

tgccgaccgc acaaggagga atgcccccca cgccgcaggc cgcgcagctc gctggacaga 1740

ggcagagtca gcagcagtat gacccctcca cggggcctcc cgtgcagaac gctgccagct 1800

tgcacacccc actgccgcag ctgcccggga ggctgccccc agccggtgtt cccactgcag 1860

ccctctcctc tgcgctgcag tttgcacagc agccgcaagt ggtagaggcc cagacacagc 1920

tccaaatccc ggtgaagact cagcagccca atgttcccat ccctgcaccg cccagcagcc 1980

aactccccat ccctccctcg cagcctgcac agctggccct ccacgttccc acacctggaa 2040

aggtgcaggt gcaggcctct cagctttcct ccctgccaca gatggtagca tcgacaaggc 2100

tccctgtgga ccctgccccg ccctgcccac ggcctctgcc cacctcttct acctcgtccc 2160

tcgcgcctgt gagtggctcc ggcccaggac cctcccctgc tcgatcctct ccagtaaata 2220

gaccttcctc agccaccaat aaggcactat ctccagtcac ttcccggacc ccaggggtgg 2280

tggcatctgc ccccaccaaa ccacagagtc ctgctcagaa tgccacctcg tcccaagaca 2340

gttctcagga tacgctgaca gaacaaataa ctctggagaa ccaggtgcat cagcgcattg 2400

cggagctgag gaaagcaggt ctgtggtccc agaggcgtct gccaaagctg caggaggccc 2460

cacgccccaa gtcccactgg gactatctgc tggaggagat gcagtggatg gccacagact 2520

ttgcccagga gaggaggtgg aaggtggctg ctgcgaagaa gctcgttaga actgtggtgc 2580

gccatcacga ggagaagcag ctccgtgaag aaagggggaa gaaggaagag cagagcagac 2640

tgaggcggat agccgcctcc acggcccggg agatagagtg cttttggtcg aatattgaac 2700

aggttgtgga aataaaacta cgagtagaat tagaagaaaa aaggaagaag gccttaaatt 2760

tacagaaagt ttccaggaga gggaaagaat tgagacctaa aggatttgac gcattacagg 2820

aaagttctct ggattcagga atgtctggaa gaaaaagaaa agctagcata tctttgactg 2880

atgacgaagt ggacgatgaa gaggaaacaa ttgaagagga ggaagcaaat gaaggcgttg 2940

tggaccacca aacagaactt tctaatttag ccaaggaagc tgagctgccc ctcctggacc 3000

tgatgaagct gtacgaaggc gccttcctgc cgagttctca gtggccccgg ccgaagcctg 3060

atggggagga cacaagcgga gaggaagatg cagatgactg tccaggcgac agggagagtc 3120

gcaaggactt ggttctcatc gactcgcttt tcatcatgga tcagttcaaa gctgccgaga 3180

ggatgaatat cgggaagcca aacgccaagg acattgcgga cgtcactgcg gtggctgaag 3240

ccatcctgcc gaagggcagt gctcgggtca caacctcggt caagtttaat gctccatctt 3300

tgttgtatgg ggctctcaga gattatcaga agattggcct ggactggctg gccaaacttt 3360

acaggaagaa tctcaatggc atattggcag atgaagctgg gctgggtaaa acagtgcaga 3420

tcattgcttt ttttgcccac ctagcttgta acgaaggtaa ttggggcccc catcttgttg 3480

ttgtgagaag ttgtaacata ctcaagtggg agcttgaatt gaaacgttgg tgtcccggac 3540

tcaaaatcct ctcatatatt ggcagccaca gagaactcaa agcaaagaga caggagtggg 3600

ccgaacccaa cagcttccac gtctgcatca cgtcctacac tcagttcttc cggggcctca 3660

ccgccttcac acgagtgcgc tggaagtgcc tggtcattga tgagatgcag cgcgtgaagg 3720

gcatgaccga gaggcactgg gaagcggttt tcaccctgca gagccaacaa cgtctgcttc 3780

tgatcgactc gccgctgcac aataccttcc tggagctctg gaccatggtg cacttcctgg 3840

tcccagggat ctccaggccc tacctgagct cccctctgag ggcccccagt gaagagagcc 3900

aggattacta ccataaagtg gtcataaggt tacacagggt gacacagcca tttattttga 3960

ggagaactaa gagagatgtg gaaaagcaac taacaaagaa atatgagcat gttttgaagt 4020

gtcgcctttc taaccgacaa aaagccttat acgaggacgt tatcctgcaa cctggcactc 4080

aggaggcctt gaagagcggg cactttgtca acgtcctgag catccttgtg cggctgcagc 4140

gcatctgcaa ccaccctggg ctcgtcgagc cccggcaccc aggctcttcc tacgtggcgg 4200

ggccactgga gtatccgtcc gcatctctaa tcctgaaggc actggagaga gatttctgga 4260

aggaagcaga tctttctatg tttgatctca tcggcttaga aaataaaatc actcgtcacg 4320

aggcagagtt gctgtctaag aaaaagatac cgcggaaact catggaggaa atctccactt 4380

cagcagcccc agcagcccga ccagcagcag caaagctgaa ggccagcagg ttgtttcagc 4440

ctgtgcagta tggccagaag cccgagggtc gcaccgtggc tttccccagc actcacccgc 4500

cccggacggc agcccccacc acggcctctg ctgctccaca gggcccgctt cgaggacggc 4560

cgcccatcgc cacgttctct gccaatccgg aggcaaaagc agcagcagcc ccgtttcaga 4620

cctctcaggc ttccgccagt gctccacgac accagcccgc ctcggcctcc agcacagccg 4680

ctagcccggc ccatcctgcg aaactgcggg cccagaccac agcacaggcc tccaccccag 4740

gccagccccc gccccagccc caggccccct cgcacgcggc cgggcagagc gcgctgcctc 4800

agaggctggt gctcccctcg caggcccagg cccgcttgcc cagtggagag gtagtgaaaa 4860

tagctcagct ggcatccatc acaggaccac agagccgcgt ggctcagcca gagacgccgg 4920

tgacactgca gttccagggc agcaagttca ccctgtcaca cagccagctc cggcagctca 4980

cagcgggcca gccgctgcag ctgcaaggca gcgtcctcca gatcgtgtcc gcccccgggc 5040

agccctacct tcgagcccct ggccctgtgg tgatgcagac cgtgtctcag gcgggcgctg 5100

tgcacggcgc cctgggaagc aagcccccgg ccggcggtcc cagccctgca cccttgaccc 5160

cacaagttgg cgttccgggc cgcgtggcgg tgaatgcctt ggctgtagga gaacccggaa 5220

cggcctccaa accagcttct cccattggag ggccgaccca ggaggaaaag accagactct 5280

tgaaagagcg cctggatcag atttatttag tcaacgagcg gcgctgttct caagctccag 5340

tctatggcag agacttgcta aggatttgtg ccctgcctag ccatggaagg gtacagtggc 5400

gtgggtccct ggatggccgt cgtgggaagg aggccgggcc agcgcacagt tacacttcat 5460

cctcagaaag tccaagtgag ctgatgttga cgctttgtcg gtgtggagag tctctgcagg 5520

atgttattga cagggtggcc tttgtgattc ctccggtggt ggcagcaccc ccgtccctac 5580

gggtgccgcg gccgccaccc ctgtacagcc acagaatgag gatcttgagg cagggcctga 5640

gagagcacgc tgcgccgtac ttccagcagc tgcggcagac cacggctcca cgcctgctgc 5700

agttccctga gctgaggctg gtgcagttcg actcagggaa gttggaagct ttagctatct 5760

tgcttcagaa attgaaatct gaaggacgtc gggtgctgat tttatcacag atgattctta 5820

tgttggacat tttagagatg ttcttgaact tccattacct cacctatgta agaatcgatg 5880

aaaatgccag cagtgagcaa cggcaggaac tgatgaggag tttcaacaga gacaggcgga 5940

ttttttgtgc cattctctcc actcacagcc gtaccacagg tataaacctt gtagaggcgg 6000

acaccgtcgt gttttatgac aatgacctga atccagtgat ggatgccaaa gctcaggagt 6060

ggtgcgatag gatcgggaga tgcaaagaca tccacatata caggcttgtg agtggcaatt 6120

ccattgaaga gaaattgttg aaaaatggaa ctaaagatct gatccgagaa gtggctgctc 6180

agggaaatga ctactccatg gctttcttaa ctcagcgaac catccaggag ctgtttgaag 6240

tttattctcc catggatgat gctggcttcc cggtcaaagc tgaggagttt gtggtgcttt 6300

ctcaggaacc ttctgtcacg gaaaccattg cacccaaaat tgcaagacct ttcatagagg 6360

ccctcaagag tattgagtat ctggaggagg atgcccagaa gtccgcacag gagggggtgc 6420

tgggaccaca cactgatgct ctgtcatcag actctgagaa catgccgtgt gatgaagaac 6480

catcccaatt agaggagcta gctgacttca tggagcagct tacaccaatt gaaaaatatg 6540

ctttaaatta cctggaatta ttccatactt ctattgagca agaaaaggag agaaacagtg 6600

aggacgcagt gatgactgca gtgagggcat gggagttctg gaacctgaag accctgcagg 6660

agagggaggc ccggctgcgg ctggagcagg aggaggcgga gctcctgacc tacacgcgag 6720

aggatgccta cagcatggag tatgtctacg aagatgtcga tgggcagaca gaagtcatgc 6780

cgctctggac cccacccacc ccgccgcagg acgacagcga catctacctc gactcggtca 6840

tgtgtctcat gtatgaagcc actcccatcc cagaggctaa gctgccccct gtgtacgtga 6900

ggaaggagcg gaagcgacac aaaacagacc cctcagctgc aggcaggaag aagaagcagc 6960

gtcacgggga ggcggtcgtc cctcctcggt ccctgtttga ccgcgcaaca ccaggacttc 7020

tgaaaattcg cagagagggc aaggagcaga agaagaatat tctgctgaag cagcaggtgc 7080

cattcgccaa gcccctgcca acttttgcca aacccacagc tgagcctggt caagacaacc 7140

ccgagtggct catcagtgag gactgggcgc tgctgcaggc tgtaaagcag ttactggagc 7200

tgcctttgaa cctcacaatc gtgtcacctg ctcacacacc taattgggat cttgtcagtg 7260

acgttgttaa ctcctgtagc cgaatctacc gctcttccaa acagtgccgg aatcgctacg 7320

agaatgtcat cattccacga gaggagggga agagtaaaaa caaccgtcct ctccgtacga 7380

gccagatcta tgcccaggat gagaatgcca cacacaccca gctgtacacg agccactttg 7440

acttaatgaa aatgactgct ggcaagagga gtcccccaat caaacctctg cttggcatga 7500

atccctttca gaagaacccc aagcacgcgt ctgtgttggc agaaagtgga atcaactatg 7560

acaagccgct gcctcccatc caggtggcat ctctccgtgc agagcgaatc gcaaaagaga 7620

aaaaggctct ggctgatcag cagaaggcac agcagccggc cgtggcccag ccacccccgc 7680

cccagccgca gcccccacca cccccgcagc agccaccgcc accgctgcca caaccacagg 7740

cagcgggcag ccagccgcca gcagggccac cagctgtcca gccccaaccc cagccacagc 7800

cccagaccca gccacagcct gtgcaggccc cagcgaaggc gcagcccgca atcacgacgg 7860

ggggcagtgc agccgtactg gcaggaacca ttaaaacatc agttactggg acgagcatgc 7920

ccactggtgc cgtgagtgga aatgtgatcg tgaacaccat cgcaggggtc ccagctgcca 7980

ccttccagtc catcaacaag cgcctggcgt cgccagtggc tcctggggcc ttgactacgc 8040

cgggaggctc tgctcccgcc caggtggtgc acacccagcc cccgccacgg gcagtcggct 8100

ccccagccac ggcgacccct gacctggtgt ccatggcaac gactcagggt gttcgagcgg 8160

tcacttctgt gacagcctcg gccgtggtca ctaccaacct gaccccagtg cagaccccgg 8220

cacggtcttt ggtgccccaa gtgtcccaag ccacaggagt tcagctccct ggaaaaacca 8280

tcacacctgc acatttccag cttctcaggc agcagcagca gcagcagcaa caacagcagc 8340

agcagcagca gcagcagcag cagcagcagc agcagcaaca gcagcagcag caacagacga 8400

cgacgacctc tcaggtgcaa gttccacaga tccagggcca ggcccagtcc ccagcacaga 8460

tcaaagctgt gggcaagctg acgccggaac acctcatcaa aatgcagaag cagaaactgc 8520

agatgccccc gcagccccca ccgccacagg cccagtctgc gcccccgcag ccaacagccc 8580

aagtgcaagt gcagacctcg cagccgccgc agcagcagag cccccagctc acgacggtca 8640

cggccccaag gcctggtgcc ctgctgacgg gcaccaccgt ggccaacctc caggtggccc 8700

ggctcacccg ggttcccact tctcagctgc aggcgcaagg gcagatgcag acccaggcac 8760

cccagccagc ccaggtggcc ttggcgaagc ctccggtggt gtccgtcccg gcagctgtgg 8820

tctcctcacc gggagtcacc accctgccca tgaacgtcgc ggggatcagc gtggcgatcg 8880

gtcagccaca gaaggcagca ggacagaccg tggtggccca gcccgtgcac atgcagcagc 8940

tgctgaagct gaagcagcag gccgtccagc agcagaaggc catccagccc caggctgcac 9000

agggcccggc agccgtccag cagaagatca ccgcacagca gatcaccacc cctggcgcgc 9060

agcagaaggt tgcctacgcc gcgcagccgg cccttaagac ccagtttctt accacaccca 9120

tctcccaggc ccagaaactg gccggggccc agcaagtgca gacccagatc caggttgcaa 9180

aacttcctca agttgttcaa cagcaaacac ccgtggccag catccagcaa gttgcctctg 9240

cttcccagca ggcttctcca cagactgtgg cgctcacgca ggcgacggcg gccgggcagc 9300

aggtgcagat gatccctgca gtgaccgcga ctgcccaggt ggttcagcag aaactcattc 9360

agcagcaggt ggtgaccacg gcgtcggccc cgctccagac tccaggcgct cccaacccag 9420

cccaggtgcc cgccagctcc gacagcccaa gccagcagcc caagttacag atgagggtcc 9480

ctgctgtcag gctaaagaca cctactaagc ctccgtgcca gtagtcaggg cagcagggct 9540

gcctctcatc taaagcaaaa ctaccttcct cacagaaaac gctttattag tgaaccttgg 9600

gaccatgtca cgcaagagat tcagcactgg gaaagatata attgaaacaa aatagtgtaa 9660

tcattttatt aaaatgcatc ccacactgca ggacaaatgg tccttatgga gtgccgcgtt 9720

ctctgtacta cgtggctcat ggaaaaagtg acaacatggc ttcctctaaa tcatttcacc 9780

tttcagtccc cacccgcacc cgtcccctag agccatagta ctgtgttctg aaagccattt 9840

agaatttctt tgtgagcatg tagtgctttg cacgccacag aagccgtctg ccgtgtgtga 9900

ggagcataca atggactttc taaagataag gcgtgggctt ccacagtgtc tgccagagtt 9960

tagttcttta taccttactg aaaaatgcct cgtggtcttc gcagagggga aggcctgtct 10020

aaagtcaatc atccgagatg ggttttccat tccaaagaaa ggcaatatgg ttccttcctt 10080

ccctcctaaa atatgactta acttttaaga gaaatgttct gacacccacc taaacacaca 10140

aggcacgttc ctggcctgtg ttcaagggaa atgatcagtc attgcattgt tattccaaag 10200

agcagccaac agtggcctcc cccaggccct accctgcaat gggattcgct ttcatttaat 10260

ggaaacttct gggactgatg cccaactcag tgcactcaag acgcatctcc agttttcggg 10320

ggaagctggt atttgacata gtgtgttaaa cagctcctga gaacctttgg gacactctgc 10380

catggctggc gtgaggccca gaggaccacg cagaggcaat ggtagtacag atgtcacagc 10440

tgagggtacg atgaggcctg ggctcagtga gccaggacga atgtgacaga caccccttgc 10500

tgccacagtc agccctttga cgaaggtggg ctggtgattc tggaagtatt ggctatagcg 10560

gtgggcccag tcaactcttc cttgtggact tacgacagca gattttctct aggataagct 10620

tgtgtggttc tgccagtgaa gcagagaacc acctgtgctg ttgtggaagg cgtgccgttg 10680

agggggaaaa cgaagcccag tatttgctac tgtttttcct ttttttacta tgacaggaaa 10740

ataaatgcaa ttttagtgga attgattgac agtgtctcct tactttgaag ttttcaccaa 10800

agcaaaaagg tccatatcca atagtatcct ttgtgctgtg gcttgatttt ggcctatttt 10860

acattatttg gtccaggaaa ttaggttata ttaggttttt tgtatactaa aaatcagtta 10920

tggcacaata aagattttct gtttttaaat tgtatttcat ctgcttcctc cccattctct 10980

cactttaagt gacattgagg aaggtattct gtcccacagg tttctgtgga cagcgataca 11040

gcaggagtca gtgaaatcaa ctggggagct cacttgagct cttgataaga aatgtggaga 11100

aaagtaaaaa ccaagctttg aagaaacaga agaaattaat cttttagtta gttgaacata 11160

ccaaagcaga ggactggaat ctgtttgttc taaccaaccc gttctccctg gcttggcacg 11220

tgccgtgaga gcgcagcttg ccggagggag ggccgctgtg tgcgcctcac atctggctcc 11280

cagtggaaac ttttactcct cctcatccgc agatgtgata gaactgaagt atctaggaat 11340

tctgcctttg tcatttgttt taatttgtgt gccctgttca ttttttttgt ctttcccaaa 11400

tcttggtagt ctccttatag ttgaagataa aatgttgagt gcacttattt tagaatatcc 11460

tagacataac tgtctaagta aaagcgctct attaatctaa aacactacaa gagaatttaa 11520

caccatctct caaatgcttt tttggagagc ttaatgggat tctgaatatt tgcaatgtgg 11580

agtttccgcc ccgatctcac gtcagtgagg gtctcctgtc tctcaagtgt gtttcctttg 11640

gctgttccct aatacaaaac acggacatat ttttactcgt agcactcaat ttagtaactt 11700

ctagatgcta ccgttgacct gagttaaatt catttagtcg tgtacgtaaa aactctcctt 11760

ttagtgtgtt attttcttgg ccttcccttt taaaggttaa agtttctaac ctaagaatta 11820

agtacgcgtt caggaagctg ttgtctaggc cttccccttg tgaatctggg ttcattccaa 11880

tacggcaagt aagagttgga aactttgaga acacagacta taaaggcagc agcccgaaca 11940

ctgtcagact ctaattggcg accctgggaa acagttgccc tgctattctt taaagaaaga 12000

cgtttattct gatgataaaa acagttagcc agactgtttt taaagcacct ggcgggaagc 12060

agaaggttgg atccaagccc ttgttcagat ttggtgcctg ataagacagg ggtttctctt 12120

tttgtgacct ttattattat tattttgtta actgttgtaa ccagttagct gttgtgtttt 12180

aagatagaaa ggaacaagac taaaattgta aatactttgt aaacatcagc atttgtactt 12240

gaatagtagg attttaaagg gcattgatag cataccaaac aaaaggcaaa ataaagtgac 12300

ctttttatat atttttt 12317

<210> 2

<211> 6469

<212> DNA

<213> genus, species, Homo sapiens (human)

<400> 2

ggcgcttcct gttccggcgc caggaggagc cgcgcgctgc tggtgctgtt gccgccgctg 60

ctctagctgc cgtcagtcag gctgcgcccg cgtcttcagg gcccagtccc tcggacccat 120

cgccgcttct agaccctact gcggtctcgg atattgccgg gaaaatgtct gatgaatttt 180

cgttggcaga tgcactacct gaacactccc ctgccaaaac ctctgctgtg agcaatacaa 240

aacctggcca acctcctcaa ggctggccag gctccaaccc ttggaataat ccgagtgctc 300

catcttcagt gccatctgga ctcccaccaa gtgcaacacc ctccactgtg ccttttggac 360

cagcaccaac aggaatgtat ccctccgtgc ctcccaccgg accacctcca ggacccccag 420

caccctttcc tccttccgga ccatcatgtc ccccacctgg tggtccttat ccagccccaa 480

ctgtgccggg ccctggcccc acagggccat atcctacacc aaatatgccc tttccagagc 540

tacccagacc atatggtgca cccacagatc cagctgcagc tggtccttta ggtccatggg 600

gatccatgtc ttctggacct tgggcgccag gaatgggagg gcagtatcct acccctaata 660

tgccatatcc atctccaggc ccatatcccg ctcctcctcc tccccaagcc cctggggcag 720

caccacctgt tccatggggc accgttccac caggagcctg gggaccacca gcaccatatc 780

ctgcccctac aggatcgtat cccacaccag gactctatcc tactcccagt aatcctttcc 840

aagtgccttc aggaccttct ggtgctccac caatgcctgg tggcccccat tcttaccatt 900

aagttaacaa tggacgaaga gatgacgctt tgctttttga agtacatgta tatgcacatg 960

aatgcatata taaaaattgc tggtttcact attagagggc attcatgaaa gaacaactct 1020

tgcacctctc agagaagata actgcctctt gtacttggat gcgtagtaca tcatatgtat 1080

acaatcagat aaaagcatag aagtaaatca ttcggatgtg atttttattt ggttttcatg 1140

gaaagttaaa gtgataaagt atattgaata gttctttgac agaatttgtt taaactatga 1200

aactacacac ttaaaaatct aagatgtgga ttattgttag aatctgcaac ttcattggca 1260

aattatttca agtatttttc tataatcact ttccccttct aaataaataa acttcgagaa 1320

taacccatca taatccaaac aaatgatgcc tcaacatttt gagctgctct gtcggacaaa 1380

taaacctggt cctcttgagg ttatattttg gatatacatt tttaaactgt cagtaattat 1440

tgtcagatgt ggagttcaat agccagccag tgttcatttt tatccttgag cttttagtaa 1500

aaacttcctg gttttatttt tagtcattgg gtcatacagc actaaagtct gctatttatg 1560

gaaactaact tttttgtttt taatccaggc caacatgtat gtaaattaaa tttttagata 1620

attgattatc tctttgtact acttgagatt tgattatgag atgtgcatat tgctttggga 1680

agagctcgag gaaggaaata attctctcct ttgttttgaa cctcaaacta gataaaccct 1740

aggaattgct taactgcaac aagtaatttt cattcccaca aaaacctgag gcagctcttt 1800

tgcccagagc gttccctgta gccaccccca ccccacttgc ccttggttct ttagaaggag 1860

cacacacatc ccttgattcc tccctgatgt ggtaaactgg cacactccag gggtctaaaa 1920

cataaaacag ttgtgtttag ggaaccttaa gtcatgcaga catgactgtt ctctttgtac 1980

aagtgtgaat caaaatatgt atctcttttt cagagtctgg ttaagctatg tcattgtcta 2040

ctgcatagtt tcctgagtct gtttgtaaag tgcttatggc taacagttca gttctgtatt 2100

tgttgacagg taaataagtg gagttgagtg ccatctttga aaaaattacc ctctagctct 2160

aacactgaaa ataataataa attgtagatc tctgcaacta agtttaaagc agtgtgactg 2220

tgttgcttaa atatcaagta ttgtttataa ccaccaaaaa aaaaaagccc tggtagtttt 2280

ttggcacctt atgtttaaat cagattctta gatttggagt agacctgacc ttgttattta 2340

ttagataaca ttttgaatgt atccattgga tttctaaaat gtattgtgaa tttctcagac 2400

aaacaggatt tatgctggag ctctgttttg cttagaaata aaatatttag tagtttattt 2460

ctgctctaat taaaatgtca agaatgccaa atgctgccag ttttttggtt tgatagctac 2520

ctccttctaa gaaagcaaaa tggttacctt tgagaggaac attcagtgtt taatcatccc 2580

ttatgttaac tagatgatag attcaagctt ttagaaatga gaaagtagaa actaatttgt 2640

taagatattt tcagactgcg gaatgttgtt agctttttct ttcacttctc ttcaaggaca 2700

ggtgttagct gtctacaata ctgttgaact ctgttgtcaa agtagccccc ttagtctaca 2760

aggcaggtag ccttggcttg aattatcaat atcaaaatgt cagttaacca tggagggata 2820

aagtaatgtg aaaagtgaga tggctgcaaa gatagctctc cttacagtta ttttggctgt 2880

cctacattgg gataagctga caaattagca gtatttagtt taacactgga gcaaatataa 2940

tttgagtagg aagaagagat agcaggtttg ggaatctata attatgaagt ccattgattt 3000

tgggagaaaa tctgttgcta aaggatttga agggccatga acacaatttg ggattattac 3060

tccctataag tataataatt ttgctagtga cccatactgt ccagtgtgcc ctaaatcata 3120

ctgctattgt actccctttg ttttcaagga ctttgcaact ggtatttggg ggagattttt 3180

tttttttttt gagacggagt ctcgctctgt cgcccatgct ggagtgcagt ggtgctatct 3240

tggttcactg caagctccat ctcccaggtt cacaccattc tcctgcctca gcctcccaag 3300

cagctgggac tacaggtgcc cgccaccatg cccggctaat tttttttttt tttttttttt 3360

agtagagatg gggtttcact gtgttagcca ggatggtctc gatctcctga cctcgtgatc 3420

tgcccgcctt ggcttcccaa agtgctggga ttacaggcgt gagccaccac gtccggccga 3480

tttttttttt tttttttaat gtaagaatgg agataaaagg gataatataa tttgctttta 3540

tattgttatt tttgtaaagc atcttttctt caattcttgt tggcattctg ggccaaaata 3600

tttcaggttg gttcggtgtg gagttaagaa aagcaggcgt tttagtggag aaatggggaa 3660

cagcatcaag aaaggctttt ttcctttttt cttttttttt tggagacaga gtcttgccct 3720

gtcacccagg ctggagtgca atggcgtgat cttggctcgc tgcaacctct gcctccaggt 3780

tcaaacgatt cttctgcctc agcctcccaa gtagctggga ttacaggtgc ccgccaccac 3840

acccattttt gtatttttag tagagacggg ggtttcacca tgttggccag ggtggtctga 3900

aactcctgac ctcgtgatcc gcctgcctca gcctcccaaa gtgctgagat tacaggcgtg 3960

agccaccatg cgtgaccttt ttttcttttt aaaagggaac aatgttgctt tcaaaacaag 4020

acatgctagg ctgaaactga tttatggaaa agactgcttg ttagcaagta tatttggtct 4080

tgagggggat acagattata gaatatgctg acatttgggc ttcagaggaa gaattttcaa 4140

atctaatgga aatagttgag gtgttcagga atgctgtttc ttggagttgg aagcttaggt 4200

tttgaaatgt tgaaaccaaa aagacaaaaa ttaaaacata gaccttaggt cgtcattcac 4260

acccggttct caagaatcaa gtggagcact tcaaagacct tggcttgtct gtcccatcct 4320

gccactttct catcttttca tgcttttgaa gacaccattt acagctctga ctcagcccta 4380

ttttgtgtaa agtaatatat tgattattca gaaatagaca atacattttt taattaccca 4440

aggactgact gttttgtgca ttttactgtt ggttgtcttc agtagagaat agtaataggg 4500

cagagaaaag tatatatttt gcctcagtca gtcccaccac cacaatggac tattgggata 4560

ttttctaaaa aaccaatcaa tttgcccatg attacctcac aaataattag tgctacctgg 4620

ggtactctca aatatacagc ttttgaaact gtagatgaaa aaagctctac tcagagtttt 4680

tgtcaagact gtgcctgggt tgaatatcag tcaattgcct acacttctaa acaataagtg 4740

ccaatgtctc aattttctca ccctgaatga tagaagctag ctttatcaaa tgccaaggtt 4800

agaaagcctg gaaataaaac ttaagcacag acattcaagt ttttgaaaag cataagccta 4860

aattcagata aatcacactg atatattgta ctatgcatag aaagttgtag gtggcgttca 4920

gggaagactt tgattttaat aaagcaatat ttagtattga agacaaacac tttttatttt 4980

cagatttctg ccaagtaaaa cagaaattgc caataaaata atcagtattt tgtaaatggc 5040

aggcaagctt ctggctgtcg aaaacatctg agtcatttat tcagtagaca atatgtcctt 5100

gatccaggtt ctttgccagc tataagggaa tccctgtcct tgagaggctc atagtctata 5160

agtaacatta cagaatttgt tagcataccc attcattatt agttttacct aaacgtgtta 5220

ggatcactac tggtggaaat tgtaaccagc ctttgggcat cttaaagggt gacatgtggc 5280

atgccttttt ttttttttaa gaatttaatg tttttcaaga ttgtagtgtt gatcagcgca 5340

acaattcaag tgtgcaaagt aacaggatag tttgcctctt cactttaccc ctggataaag 5400

gcactttcac tgcctgtcac tgatcagcag atactgactt gttgccatta agtgaacttg 5460

acttcttatg tgtgctctat gagtttgttg taattttctt cttgaaattg tgatttttca 5520

ctgacagtaa tgacaaattt aatgtatgta attgtctatg cattttaagt taaactgcct 5580

aaaatgtgat ttgagacata tacatatgtt tgtattataa attgtaagca atcagtttga 5640

gatactaggt tttatcacct gctgctgtat ttgtaaacaa agacaaatgt tgctttaaga 5700

agtaattata attaggaata ggctatggat gtgatacttg gtatttttta agataaactt 5760

gtttgctttt gtgtattata cctggaaact ttttttaaaa aatgtatttt catggtttca 5820

cagatttttc atgttatttt attctttagg cccaattctg ggcttctctg agcaagtcca 5880

gagcctaatt aactgtaaat ttgttgtcaa aaaggaagaa aaaagggcct gagatacctc 5940

tttgcatgtg acctgcattc actaaggata tctggaaacc acccttcctc cgcaaaccct 6000

ctcagcaaca tggtgtccat tgtggtgatt ttctcttctt ttaaggctag gctactcttg 6060

gtaaccagat tatccgtata tatgataata tgaagtcagg gaactttctc tgtctgtccc 6120

tactcccctc actcccccac tttctgttat gaaagatagt tctactttta tcattaactg 6180

ctacgcattt agtgagggtc acattattaa acttggagtt taccattttc ccacaggaga 6240

tttcgctggc attccttgga actcccaatt tcagtagggc aatgaatgaa tgaatacttt 6300

gcagtgctac ttttggaagg aatttctgct ttttgcctta tgattggaca aaatgcagct 6360

gtaaaatttt aaattgtttt tgatatgtta ttcaatatcc catgaaagta ttcacctaaa 6420

gtggagttat gaaatggatg gtgaaataat aagaccattc tggagcagg 6469

<210> 3

<211> 10897

<212> DNA

<213> genus, species, Homo sapiens (human)

<400> 3

agatatactg agtgagccct gagaagcagt ctcagatcct gacggtgcag cagcccgcag 60

cctcagccag ggagtcccag ccgctttcaa tggaggagaa gcccggccag ccacagcctc 120

agcaccatca cagccaccac catccgcacc atcaccctca gcagcagcag cagcagccgc 180

accaccacca ccattattat ttctacaacc acagccacaa ccaccaccac caccatcatc 240

accagcagcc tcaccaatac ctgcagcatg gagccgaggg cagccccaag gcccagccaa 300

agccgctgaa acatgagcag aaacacaccc tccagcagca ccaggaaacg ccgaagaaga 360

aaacaggcta tggtgaacta aacggtaatg ctggagaaag agaaatatct ttaaagaacc 420

tgagttctga tgaagccacc aaccctattt ccagggtcct caatggcaac cagcaagttg 480

tagacactag cctgaagcag actgtaaagg ccaacacctt tgggaaagca ggaattaaaa 540

ccaagaattt cattcagaaa aacagtatgg acaaaaagaa tgggaagtct tatgaaaata 600

aatctggaga gaatcagtct gtagataagt ctgatactat accaattcca aatggtgtgg 660

taacaaataa ttctggttat attactaatg gttatatggg taaaggagca gataatgatg 720

gtagtggatc tgagagcgga tatacaactc ctaaaaaaag gaaagctagg cgcaatagtg 780

ccaagggttg tgaaaacctt aatatagtgc aggacaaaat aatgcaacaa gagaccagtg 840

tcccaacctt aaaacaggga cttgaaactt tcaagcctga ctatagtgaa caaaagggaa 900

atcgagtaga tggttcgaag cccatttgga agtatgaaac tgggcctgga ggaacaagtc 960

gaggaaaacc tgctgtgggt gatatgcttc ggaaaagctc agatagtaaa cctggtgtga 1020

gcagcaaaaa gtttgatgat cggcccaaag gaaagcatgc ttcagctgtt gcctccaaag 1080

aggactcgtg gaccctattt aaaccacccc cagtttttcc agtggacaat agcagtgcta 1140

aaatagttcc taaaataagt tatgcaagca aagttaagga aaacctcaac aaaactatac 1200

agaactcttc tgtgtcacca acttcatctt catcatcttc atcatctacc ggggaaactc 1260

agacccaatc atcaagtcgc ttatcccagg tccctatgtc agcgctgaaa tctgttactt 1320

ctgccaactt ttctaatggg cctgttttag cagggactga tggaaatgtt tatcctccag 1380

ggggtcagcc actgctaact actgctgcta atactctaac acccatctct tctgggacag 1440

attcagttct ccaggacatg agtctaactt cagcagctgt tgaacaaatt aagactagcc 1500

tttttatcta tccttcaaat atgcaaacta tgctgttgag cacagcacaa gtggatctgc 1560

cctctcagac agatcagcaa aacctggggg atatcttcca gaatcagtgg ggtttatcat 1620

ttataaatga gcccagtgct ggccctgaga ctgttactgg gaagtcatca gagcataaag 1680

tgatggaggt gacatttcaa ggagaatatc ctgctacttt ggtttcacag ggtgctgaaa 1740

taattccctc aggaactgag catcctgtgt ttcccaaggc ttacgagctg gagaaacgga 1800

ctagtcctca agttctgggt agcattctaa aatctgggac tactagtgag agtggagcct 1860

tatccttgga acccagtcat ataggtgacc tgcagaaagc agacaccagt agtcaaggtg 1920

ctttagtgtt tctctcaaag gactacgaga tagaaagtca aaatcctctg gcctctccta 1980

cgaacacttt gttaggctct gccaaagaac agagatacca gagaggccta gaaaggaatg 2040

atagctgggg ttcttttgac ctgagggctg ctattgtata tcacactaaa gaaatggaat 2100

ctatttggaa tttgcagaag caagatccca aaaggataat cacttacaat gaagccatgg 2160

atagtccaga tcaatgaagg accagactgc ctattcgtaa cctttctgca gcattagagc 2220

catcgttcat gggggacaca aggcttttat gctcctagat cttcaacgca gcagaggaac 2280

cataagtaga atcacaggat aatatataca aatatatata tatacatata tatatatata 2340

gttatttaaa aaaggcaact gaaagtaatt agacttctta aggaatcaaa tttatttcaa 2400

gagactacac atggttattt aatctccggt actgaatagg ttttttttct tctgttagtt 2460

tttgttttta agtgtgaatg caagtgatta atgaatacag acttaacaag tgtggttcta 2520

aagttcctgc tgtcatcaac ttgggcaaca aatgacccac tggaaaggca aatccactta 2580

aaagatctct gtatcttgtt ctgtgactga agtgatacac taatcacggg gaacccagaa 2640

tgattcaaca ttttcccccc actcctccct tgatcttttt ggttttactt taattaagcc 2700

ctgcgagaat gctggataaa tgccttgaag ttagcagggt gtattttttt agcgaatatg 2760

atttgcatgt cttgccagga gttaagcggc ctctggggtg ttggggaaat actttatttc 2820

tttccattta ttttttgtgg ggcggggata ggggagggca ttgaagttct acaattctgg 2880

aatagttagt tgatggtaca tagttaactt ggcttcggtt acatattgga ctttaacaac 2940

tgaagaatct atgcgtgtca tttaaagaaa agttgcagaa caagcaattg gcttagatat 3000

acaatctgga aaaatattcc tgtgcccata ttttaatgta attgtataac tgggagcaaa 3060

aatatattct gcttttcaac tgtaggtgct ccagacttgc tctccgtcac taacactaaa 3120

tgtgctgttt tccttgtttt tcatcaaaca tttaagacaa acttagacct ttctgtaaat 3180

tatcttttaa tttctcagca aaatctaaaa ggggaagaaa aaagtccatg aaaactaaaa 3240

cttttcatgt ttttagccag tgagaagata ataaaccctg actgtagaag gtgtgttttc 3300

atgcaaacta tacttctgag cttgttagct tctaattata tcttaataaa tatattttat 3360

tactagagca agatgggttt ttaaggaaaa taatgtgaaa ttctggaaat tttctttggg 3420

gcagagaaga gcattagccc tgtcttatca ttacattgcc atcctgttgc actgcagctt 3480

gtgtatagca tgctaaaata aatttttgtg tgtgtgtgca gaaattaagg gtccaattga 3540

gattgggtga tgttagtaac ataataacaa gttgtctggc ctgacacagc atcacatcac 3600

acacacagaa attagtatat ccatgtatgt caaatacagg ttaaaatatc agggcattta 3660

tataaagagt tgtagtcttc tgataaaagt agactggatc ccctggggta tttggggaga 3720

aagtaactac tttggctcta cccctagaaa tgtccagttt tgagtgactg tagtatggat 3780

gggttttctt gttttgttga ttatttgagg cttttaaaac aagtagttca tgaaagaagc 3840

tgttggactc aacatagagt agagtaacta tctttttagt ctggatttct gccctgctta 3900

gattttaaaa gtataagcat ggattgccaa ttccacttga tgtaaacaaa actttttttt 3960

atacataata tatatatata tatataaaat aacttattgt atcagtccag gttcagaaac 4020

ttgtggtagg ccagttccag atagtttcat ttcacctgta aactgtatca ctttgactga 4080

tattgtaatt ttcaaatgta taatatgttt acagatgtgc cctgcattta gtctgccttg 4140

ttcctatttt gatttttgtt gagtctcctg cctgcttgcc aaaagctagg atgcttcagg 4200

cccatgtaca attgaaagca gaggcatcct tgagctttaa agcattgaac aaactggaaa 4260

atgcaacata ccacataact gaagtgaaaa aagtctgtgt ttttgtgttt ttttaaataa 4320

aaattttcaa aaagttaaaa aaaaagacat ataaggttga ttaaagggaa aaaaggctcc 4380

agtttgtttt acaggtttta aagttctgct gtgtgttcaa ttgccttgtg taaccacttg 4440

tcgccttagg gccagattcc cctctctagt cccctttttt aaatgtccat tttgcttgcc 4500

tggaatttta aagttcttcc gtctcacaac tcacaagaaa ctttctgggt ttgtgacata 4560

cagaggttga attgagtata tatttgaaaa ggaaaaaaca aaaaacaaac ccagacccca 4620

cctgaattgg gctttttaac ttagaagcaa cacttgatta aacatcttta gaaagctatt 4680

gcttttctaa tttccttcca tatccctcag gcctcagtgt tcagagaagc caaaaagaat 4740

gtatcacttc tctgtctgtc caaaggtttt tgagagtctc acttctaaat gaaacaatgc 4800

aacatttcac tttgatttct ccactgaaat ttccttgatt atatggttag aggtatgtag 4860

ttaggaatgt ctgttaactt tctgagaacc ctagtgcccc atcatattaa ctgtcagtat 4920

tttgggggca ttaggttaat agacttaatt gcctaggtac aagcaggact ttgggacaaa 4980

tctctttgtg ctgtttggta acacttaact ctatttgttg caatctttct ccttaggtcc 5040

tcacacaatt ccttacagag cacttattaa aaaaaaatct taagagttga tctgttttct 5100

gattattttg tgtaagcttc taaacaaact tcagctgtga ttaatttagc acatttaaat 5160

aacgtgttat tgtttggtat aaagaatttt ccttcaactc agagtattag tactgtagca 5220

taaaccaaat acagtctaga ggggattttt aacatccctc cattataaag actgaaaaag 5280

gggtgtgtgt gtgtgtgtgt gtttatgtat gtatgtatgt gtgtgtgagg aaaagatgga 5340

gatattaaaa attagtaaat gaatgtgtat aagacattag tattcagaga atgaacttgt 5400

atttattttg tgccatttgt tttcattaca cagaaaaaag tcaggtggtt taaatcctta 5460

aaagggtagt attgaaaaat ggcactaaga atgaaattat gacctatttt tttaatagct 5520

atgaagatac taattatggg tgaagatttc tttttaaatc tgttttgatt attgtaggct 5580

tctgtgtcac ataccactct tgtaggtgtc ctcaataatc cccttttccc acaaaataca 5640

cagggtgtat tatctttctc tttattcacc cccactttgc tgaactgaag ttaattacat 5700

agcctttctt ctaacctcct tagtaatgaa ccttcacata aagtgtattt acagcgtctg 5760

tggtagccag cccttcctcc tctactttct aggaggggat agccaataac taggaattta 5820

atgacagatt tttttttctt tgaaataaat ggccagagtt tctccatttt agaattttgt 5880

tgtcctcctt aatcatctgc ttacctagtc attactcaat ctgcagaaac ttcataaagg 5940

aaaagtgctg cattgttttt acaaataaca gtttgtaggg aaaatatgac aaacctcaac 6000

tatgggagtt gtccacaata caaaattttg aaaaaacatt acatagtgat aatatcatac 6060

ttggttgtta ggcttgttgc ttccccacat cagaggcatc taatgattta tcttttgtaa 6120

ttgctgtgaa cttttttaaa taagccattt agtgtgaaat tgtcatgtat caaatggcta 6180

ttggaaatgg actttactca attttaattc cactgtaaat aaggacggag tcattcctac 6240

aaggctctct tcagagaaat agattaaaag tccaatttcc aggtattatt agtatagtta 6300

tgccgctggg ccacatcctc aacaacagct gatccctctt gtataaatat gttaactgtg 6360

cagaacagtt atgttatggg acaaatataa tggtcattat ggtcagattg gttgatgcca 6420

caccagtcaa ggtagagtct gatagggcag tatcttaata accctaccca tgacttaact 6480

gttggatttg aaaggaaaac gtaggatttg ctcttgtccc cttacccgcc acaaaatttt 6540

gataatttgt ttaaaaggga gaggcagagg aaaagactag aagcataaat agctgcttta 6600

ggtttgccag aggcacatag cttaacatta gttcttaata tcgatgttat ttttactaat 6660

gtaattaatc aacagagcac caagattctt tcatggtgaa aagggtgggc ttctgttttg 6720

gtatcttaaa atgtttcttt taaaatatac atcacctgtg tgagaaccag gaccacctgg 6780

gagagtgatg aatcattggc tccactcaaa agcattgctt tactgagttt taaatttcac 6840

actgttttgc cgctcaagaa aggtcttaaa gtagttaaag gatgccagca atagtgcgaa 6900

tagaattttc ggttgtctgc ataataaaaa cacccattgc agcatgattg gtatgttgct 6960

cttgcattat tgggaatggt aaatcagtta tgggctagaa actatggaat ggccgtcctc 7020

atatgtgatg ggattgctga ttcagacttc cctattttcc atacaatttt gttatgtgca 7080

gagttctaaa gccattttat aatactgcag tatccccccc ccccccacct tttttttttt 7140

gagacggact gtctgttgcc caggctggag tacagtggcg caatcttggc tcactgcaac 7200

ctccacctcc ctggttcaag caattcccct gcctcagcct cccatgtagc tgggattacg 7260

ggcgcacacc accacacctg gctaatttgt attcttagta gagacggggt ttcaccatgt 7320

tgaccagact ggtctcgaac tcctgacctc aggcaatctg cccgcctcag cctcccaaag 7380

tgctggaatt acaggtgtga gccaccgtgc ccggccaagt atctcttttc tacagcctta 7440

ttaaactaac tacaaacatt tattttccaa tttagtttta ctttcagtgc atatcaaagt 7500

tgttgtactc ttcagaccaa caaattaact tgagggcaaa ttacatagct ttccatgtac 7560

ccttttttcc tcaggtgcta atcaaaggct ctgaaaatgg atactgcttt agtgatgtct 7620

gctttattct taaaatgctt atttcttttg ctagatgtaa agatttggtg ttaacaaaag 7680

tggttttaat atgtaaatat gaatgaatgc ctttagttta ccctgtttgt ctattattaa 7740

tctgttttca tttatccttc atagaggagg atcctttcat gatcttgaat acatttcatt 7800

agatattgtt gcattttaag aatgaaaata caactgtttt ctgtcttaga ttaatcctgc 7860

tgctatgaga aactgaaaat caagaatgtg atgcactttt tacattacta tataccatac 7920

atataccata ggttgctttg atacctttcc tgtagcacag ccactaacaa gagtgaatga 7980

attataaaat tctttttggg agggaatcaa tacaagtaac taattcttag ctgatattgt 8040

cctatgaagg acaataactt aggaatataa gaattctgtt aatagtacac tttttggcct 8100

taaatgtctt ctactactga aaatagttta aatcttagct ttgtttctat tattccctct 8160

ctctgcctca gaaagaggaa ttgggaagaa tggcttaaag gacgtggtgt cattgatttg 8220

ttgctgatct tttagaaaac atttgtctat gtaagctggg gacttatttt ttgtttgtat 8280

atagagggga aatagtgctg ccctgaacca atcagattta gtttaaatca aatcaatcaa 8340

aactccagct gtttctcttg tctttttact tagcaaagga aaactttagt gaatgctact 8400

tgacaagaag aaaagtcatt tctcaagcac atacccaaac ttgaaggtga ttgaacccaa 8460

aataatgggt gggaaacacc aaatgaggtg gaggaatgag aaagatgtgt gggccaaagc 8520

tatctggtta tattttgatg ttgccaatat cgcaaagcca aaattttaat ttgcttattt 8580

aatatatttg ttggccagag atctattttt atatcaatgt gccttgcatg tatattaaaa 8640

aaaaaaaatt ggaaacgcca tgtagtaatg cctgagatag tcgatggttc ttaccacctc 8700

actaattttt atgcagtatg aaatgctcat tctattgccc aactggtgct ctctgtttaa 8760

agttacagat cttgcgaaac tggaactatt ttataagctg gggaagtgat ttactttttt 8820

tgttgtatct tttttgttct tagtctgtta gtggctgtcc tgtagtggga aatagtaaaa 8880

ggattcttca ctcccttctc ccctcagcac cttcttcaag taaacatttc ttgtgtgctt 8940

tgaaaaaagt ttcagcttgc tgtctctttt agtgttttaa agaagtgtta tacaaagcat 9000

tgtttgcaaa atatagggag ataatggagt ccactttaat ttggaattct gtgtgagcta 9060

tgatccaagt tatcagctct ttccaacttt aaaaattttg ttaaaagcac cttgcttaga 9120

aaattttaaa tatttatgtc tgcaacaatt gtctcaaaat aataaactgt gcaattcttg 9180

tcattaaaaa aaaaaaagat ctgaattttc cctaatgtga cttgttagtt tctctctgta 9240

tttcctgcca gtgtaaatgt gaaagctttg cttgcattac gttttagaaa tgcattttgc 9300

acactcgaat tttgccgaag ctccgtgaaa aggttagatc taagtagatg aataaagcta 9360

tgcacatgtt ttgaaagttt aatttgtgtg tcattaccaa aagtgaccga tttgtcctta 9420

ctactttgct gttgttagct ttaccatctt tggaaacttg gctcaaagtt acatagttct 9480

gggctagctc atcagtggaa ctaggagaga ggaaaactgg cacctatttt aataaagttc 9540

aatttaaacg agagcttgac ttgtatctat taaagagctt ttcttgaaac agggcagttt 9600

tatcagcttt acaaatcatt ggatgctctt ccttagtaat attttggttt atttgatcaa 9660

atagaaatgg aaagtaattc aaactgaaag accctttttt gtcatatgga acttggtgac 9720

gattttttgt cttaaagctg gtttaaaggt aggataggct tttaccttta ttgctttagc 9780

ataaatttgg tttactgaat tgactggctt gagattagaa ttattcagtt gtttgtaaga 9840

tcaaagcact ggttgtttta aagataacgt gtatctttta aaaaattgcc caagctgatt 9900

agaacaagtt taggagttgg gtacatttgg ttcaagtgct gcaatctgta tgtactaaat 9960

agctttactt tgtgtatgtg tacttataat gtgtagatgt actactaccc aggttttgtc 10020

aaatcatctt ttttaaagtt tttttttttt taattggttc aggacctttg taggagaggc 10080

taatatgttt aagtagaaga tattactgat agcattttcc ccatgctcct acataaaaaa 10140

taaatatttc cattttatag ctttttcaat atacagaaga gggttacttc ttcatcaagt 10200

atattgttgc ctttgaggac acagcaaaac ccttctatat gtatcttcat tgatagtggc 10260

agttaaaaac taagttatcc agttaagact taaaaggtga cccatattaa ttgcatggcc 10320

ttaaaaggca gaaatgcagg agtgtagcaa gcatcatttt agatggctat ggttcctctt 10380

ccgcatctgt cagtagttca cttatgttca gtcttagaac ctactggagg agtgaagtaa 10440

tttctctgtc tcgtgcagag gcactaagga gctgagttac ctcttaatct gggggaatgg 10500

ataataagtg gagtacagtt atgttaaagg atgttccccc cgctcaaaaa aaagtttcaa 10560

tgtttgtttt gcccagtcaa aaatataggt cttttctaca tataagaaca gtcaccagaa 10620

attttccctt ttgctaaatg cttaggtatt tgctatagct gtttctgatg tcatggattc 10680

tgaggaagtg tcatttacgt gatgatcttc ctttattgat gtcttcatca tgttcagtgt 10740

tttaaaaata taaattacaa acactctaca accataccca gatttactta ttttatcaga 10800

aaaaaaactt gagaaatttg tagatcaaat tgagagacaa taagtgtaca ttgttgaata 10860

aaaaatttta aagtttctga aaaaaaaaaa aaaaaaa 10897

<210> 4

<211> 12687

<212> DNA

<213> genus, species, homo sapiens (human)

<400> 4

atgcgcagcc catgttagtg atggaggaga gaagatggcg gaagcggaat ttaaggacca 60

tagtacagct atggatactg aaccaaaccc gggaacatct tctgtgtcaa caacaaccag 120

cagtaccacc accaccacca tcaccacttc ctcctctcga atgcagcagc cacagatctc 180

tgtctacagt ggttcagacc gacatgctgt acaggtaatt caacaggcat tgcatcggcc 240

ccccagctca gctgctcagt accttcagca aatgtatgca gcccaacaac agcacttgat 300

gctgcatact gcagctcttc agcagcagca tttaagcagc tcccagcttc agagccttgc 360

tgctgttcag gcaagtttgt ccagtggaag accatctaca tctcccacag gaagtgtcac 420

acagcagtca agtatgtccc aaacgtctat caacctctcc acttctccta cacctgcaca 480

gttaataagc cgttcccagg cttccagttc taccagcggc agtattaccc aacagactat 540

gttactaggg agtacttccc ctaccctaac ggcaagccaa gctcaaatgt atctccgagc 600

tcaaatgctg attttcacac ccgctaccac tgtggctgct gtacagtctg acattcctgt 660

tgtctcgtcg tcatcgtcat cttcctgtca gtctgcagct actcaggttc agaatttaac 720

attacgcagc cagaagttgg gtgtattatc tagctcacag aatggtccac caaaaagcac 780

tagtcaaact cagtcattga caatttgtca taacaaaaca acagtgacca gttctaaaat 840

cagccaacga gatccttctc cagaaagtaa taagaaagga gagagcccaa gcctggaatc 900

acgaagcaca gctgtcaccc ggacatcaag tattcaccag ttaatagcac cagcttcata 960

ttctccaatt cagcctcatt ctctaataaa acatcagcag attcctcttc attcaccacc 1020

ttccaaagtt tcccatcatc agctgatatt acaacagcag caacagcaaa ttcagccaat 1080

cacacttcag aattcaactc aagacccacc cccatcccag cactgtatac cactccagaa 1140

ccatggcctt cctccagctc ccagtaatgc ccagtcacag cattgttcac cgattcagag 1200

tcatccctct cctttaacag tgtctcctaa tcagtcacag tcagcacagc agtctgtagt 1260

ggtgtctcct ccaccacctc attcaccaag tcagtctcct actataatta ttcatccaca 1320

agcacttatt cagccacacc ctcttgtgtc atcagctctc cagccagggc caaatttgca 1380

gcagtccact gctaatcagg tgcaagctac agcacagttg aatcttccat cccatcttcc 1440

acttccagct tcccctgttg tacacattgg cccagttcag cagtctgcct tggtatcccc 1500

aggccagcag attgtctctc catcacacca gcaatattca tccctgcagt cctctccaat 1560

cccaattgca agtcctccac agatgtcgac atctcctcca gctcagattc caccactgcc 1620

cttgcagtct atgcagtctt tacaagtgca gcctgaaatt ctgtcccagg gccaggtttt 1680

ggtgcagaat gctttggtgt cagaagagga acttccagct gcagaagctt tggtccagtt 1740

gccatttcag actcttcctc ctccacagac tgttgcggta aacctacaag tgcaaccacc 1800

agcacctgtt gatccaccag tggtttatca ggtagaagat gtgtgtgaag aagaaatgcc 1860

agaagagtca gatgaatgtg tccggatgga tagaacccca ccaccaccca ctttgtctcc 1920

agcagctata acagtgggga gaggagaaga tttgacttct gaacatcctt tgttagagca 1980

agtggaatta cctgctgtgg catcagtcag tgcttcagta attaaatctc catcagatcc 2040

ctcacatgtt tctgttccac cacctccatt gttacttcca gctgccacca caaggagtaa 2100

cagtacatct atgcacagta gcattcccag tatagagaac aaacctccac aggctattgt 2160

taaaccacag atcctaaccc atgttattga aggctttgtg attcaggagg gattggagcc 2220

atttcctgtg agtcgttcct ctttgctaat agaacagcct gtgaaaaaac ggcctctttt 2280

ggataatcag gtgataaatt cagtgtgtgt tcagccagag ctacagaata atacaaaaca 2340

tgcggataat tcatctgaca cagagatgga agacatgatt gctgaagaga cattagaaga 2400

aatggacagt gagttgctca agtgtgaatt ctgtgggaaa atgggatatg ctaatgaatt 2460

tttgcggtca aaacgattct gcactatgtc atgtgccaaa aggtacaatg ttagctgttc 2520

taaaaaattt gcacttagtc gttggaatcg taagcctgat aatcaaagtc ttgggcatcg 2580

tggccgtcgt ccaagtggcc ctgatggggc agcgagagaa catatcctta ggcagcttcc 2640

aattacttat ccatctgcag aagaagactt ggcttctcat gaagattctg tgccatctgc 2700

tatgacaact cgtctgcgca ggcagagcga gcgggaaaga gaacgtgagc ttcgggatgt 2760

gagaattcgg aaaatgcctg agaacagtga cttgctacca gttgcacaaa cagagccatc 2820

tatatggaca gttgatgatg tctgggcctt catccattct ttgcctggct gccaggatat 2880

cgcagatgaa ttcagagcac aggagattga tggacaggcc cttctcttgc tgaaagaaga 2940

ccatctcatg agtgcaatga atatcaagct aggcccagcc ctgaagatct gtgcacgcat 3000

caactctctg aaggaatctt aacaggaaca tgaagccttg ataaaacagc agttttactt 3060

ttctcacaaa aacttgtaag gtaaaggcct aacttggtct agaatatgac acttattgtg 3120

gtggatagcc aagcacattg ggatctccac atcaaatact gacatttctt ctacaggtat 3180

aataattcat catgcatttt cataattaat aaacattggt aaaattaatt ttacaggtta 3240

catgaaacat tgaaagactt gttacagagg gccatgatat ttttcaaaga aatgtgttat 3300

actagataat ttttttaaag gtgatgttta tcattaatat aaagaatcct tttaaaagta 3360

atttaatgat ttacatttct cctcttttga ttcaattttc ttatacattt tttctaccct 3420

attagttttc taaaggttgt catgagaggt atattatgga ataatttagt agtccagtga 3480

cagaatcgta tgaaatcagt gtacatttta aaaaacatgt cttttagaca tatgctttat 3540

ctataaaaaa ggaattgtgt tctagtatga acaatactga tctggaagtg agaagagtta 3600

gtttctattc caaacttgac caagaatttg gtttgactga gaacgttttc ctctcagttt 3660

ttgtacattt atttagagca gtggttctca gtggaggtca gttttgatcg ccaggggaca 3720

tctggcaatg ttgagacatt ttggttgtca cagcttgggg gtgggttcag gggagggttg 3780

ctactggtgt ctagtagtta gaagccagag atgtttctaa acatcttata atgcacagga 3840

cagcacccct ccactgtaaa gaattattgg ttcaaaaata tcggtactgc caaggttgag 3900

aaactctgat atagaaggag tgataaatat tgttttcacc caaaggaata cttttaaagg 3960

atgaagctta ctaaacatat atgatggaag tattattcag ataacattaa tattctgctg 4020

aataattttt tctagtttaa tcatactaga aaaagaaaaa aaatctacaa attgtcctat 4080

aaaataagga caaacatgca aataatttaa ctctcagaaa gtactaattc attctgatta 4140

tctttcatac ctctgtgctc ctctgcactg acgaagacat aatatgatta tacctatgaa 4200

ctagtgcaca gccttttctg gcaagaaaat agtttgtagc agatacgtgg ttgctctttg 4260

gatttttttc tattgttgaa catgctggga ctagctagaa tgcacattcc tacttccttt 4320

accaaacgtt tgcatgcttc ctgcaaagca cttaccaagt gatttctctt gaaccatcgg 4380

atataatttt gtatgtacat gtttgaggaa aaaaatgtaa agcaaaacct tttactgaac 4440

agtgttctat agaattatga cactaaaaca aaattgtttg tggaagccct gaaagcttta 4500

tagtcctgga catcaaaaat tttatttgag atgatgaatg ttttgttttc atcttttctt 4560

atattaccac aattgagata ttttagtaat tgaaggaaca tacacagata tttggcagaa 4620

gtcgagtaag gaggggaaaa aaagagtccg tgagtttcag tcattttcac tgctcttttc 4680

aaaaagattg tgttgagctg gtagaagact aaagatgtca ctgaagacat cacagatact 4740

atatttatct tttggctttg tgtacattag agaatgttga ttatttttat acaaaaatac 4800

agcgggtaat ttttttaatc tttagatgcc tcttgtttga atgtatgctt tgtggaattc 4860

tttgtgtagt aatgttttaa aaaaagatgt ttactgatag ttacatgtag gattagaata 4920

tgtaatataa tataaggctc atgttccaga cctacgatag cttgtagtct atgttacgta 4980

tttctttata tcacattttt aatcattgga ttaaagtatc aaggaaagct aggtactcta 5040

taatgagttt tcatttatta gcagttaatc atcatgacag aattgtcata tgcttgactt 5100

ttccctcttc ttggaatttc agaacacaaa tacaggctaa gcattagtaa gagatggccc 5160

acagtatgag agagagaggt gcaacggaaa atctcgcctg gaattaaaac ttttcataga 5220

ttatccacgg ttaatacaaa atttattata tggggataga ctgctccagc aataatgatt 5280

acatcctata actgtattac ctatggcctt taaggtatca attttgaact gtgttgtagg 5340

ctctcctttt atttgttctc tttcctaata gcagccattc tgtacttatt gaaagcccct 5400

gtgcctactg ctgtcttaag tattcaggag gggcttacaa gagggttttc tattggagaa 5460

taccgtataa tcttaaatct agtccagatc tctgttgtcc ccactcaaaa catacacaaa 5520

atatgcactt gcttttttca agtgagtttt tatttaaaaa tggcttgttt gctatcacat 5580

tggtgcagct gtttctttca agatgagtta atcatcttaa tttcaaagct tcagctatat 5640

ataatggata tatagacaac actgagcatc cacctctctc ctgagcttta aagcagagtt 5700

tcagtatgat ataggtgggg agagtaaatt gttttcatat cctttcatac tactactaat 5760

agttttagga ttttgactgg ggagagataa tgacaaacag aaagggaaca tggaggttct 5820

tcctactttt gctacctaag tttgcatttt ctgacttcct tgcagtgttg cactctttgt 5880

cccattggga taaaaagcat aagtttgaaa ttttgcttta agccttgtgt tcctggggaa 5940

gttaaacaac taagagagct gatttgtaaa aattattttt tatatgacat taatattcat 6000

caagccttgt gtaggcatgt gtaagacaca gctatgcagc tttgagtagt caatatagta 6060

tgagatagag tgttgtccca aatcctcctg tcacttttta agtagcatat tatttccctg 6120

atggtcctgt tactttgctg ttgaatgctc taaacagaac tttttaaaag gtgtgtttta 6180

agagcagtca cctaggagta gacaaggtgg aatgggagga gagaaatggt aatgcaaaag 6240

cttgagcatg ggaagagtca gaggaggagg ccatcatcct tgttagctta gcctacttca 6300

acactgagca catttctgca cttttgaagt gaaattcatg ttttacttag aagaaataat 6360

tttctttcat tagggatccc agttgatttt tgtttcctgg tgtatcaaaa tacttagaac 6420

tatgaaacaa gtattattgt gatcatgcct ttgaataatt tttgacgtag cttatcttca 6480

tgtatcaagt ataaaattat aatgagacat ctattcacaa atacaagtct tagattgaat 6540

tgaaatgtgt tatagtgccc tgtctcccac tgacttgttc agttaaatgt cttaaagtac 6600

attatgtaca tcttcaggct tttggtacca caatggcaca agtatggtag ggaggcaata 6660

tagtcttagg ctatatgcct atattaagtg tgtataaaca atttttgaaa gaatacacta 6720

ttatagatgt atgtgagtga tgctgacctg acagccatat ccagtggatg aaactgactg 6780

gacacactgt taaaatgttt taaagatgta ttttcagcca gaacagcctg gttatagttt 6840

gtggttttca ccttggtgga ttgcaggaac acatgcagcc tactggcatt gagcattagc 6900

taatggcatg aaagggcctc atctcactac ctctctaagg cctctagctc caagaaaacc 6960

atgaaaactt ctttcttgga gagatctttg tctcagaatc cttagagagg atttcgtatg 7020

ggggctaact ttaggaaggg aggcagctgg ggcaggactt tctgatacct gacagtcatg 7080

ttccagagca acctttgggc agtggaaact ggcgcatcta tgcaaaatga ttgctcaatc 7140

tctatcttgt gtactacata tgtaactagc tgggccctaa ggaaggtttt ctagggggaa 7200

ggatagggaa gtagaggagg agacaagtag gaggaacaaa gcattctaga cccaagagga 7260

tagaagatat ttaggataga tatggctttc atccatagtt caaaataatg cgttttgtta 7320

gatgccagtt atagcagtaa ataggttata gtttttatat gtcaagattt acctgtaatc 7380

agactcattc tttcactctc tatacccact gtctccatgc ttgggagcat ggatattaat 7440

agttccagtg atgtagaagt tagtgatttt tgatttctga aaaaggtgag aaccttttat 7500

tacagttgga gaatatttgt caaaaattca aaggttgttg taattgagtt gccagaatta 7560

cagagtttcc attttcagat atcacagttg aatcacctct gtagattgtt ataaagagag 7620

gcattttaag atagtatttt atttgctagg ttgtgtctca gtctaagaat tgggaaaaga 7680

agagctatag gtttctcttt cctagtctgg atttcagtaa acacaagcct acctctgctt 7740

ctttggttca cagcagtgtg gatcatgaaa tgaactgttt acccacattc atcaatattg 7800

gtattttaca aatctacttg gagcatttaa tttcatctca aagattgtga tccactttag 7860

ataagcacaa atacagtatt aggaaaagta aatatgcaat cttactaaaa tttcaacttg 7920

ttaagctgta tatcttaaaa gaaattattt ggggctgggc atggtggctc acacctgtaa 7980

tcccagcact ttgggaggct gaggtgggta gatcacctga ggtcaggagt tcgagaccag 8040

cctgaccaat atggtgaaac cctatctcta ctaaaaacac aaaaattagc tgggtgtggt 8100

ggcatgcacc tgtaattcca gctacttggg aggctgagac aggagaattg cttgaaccca 8160

ggtggtggag gttgcagtga gccaagatca cacccctgca ctccagcctg ggtgacagag 8220

cgagactcca tctcaaaaaa acaaaacaaa aaattatttg ggaagatacg tcctctttta 8280

ttagaagttc ataaaatgta tcatatagtt ttgttcacag tagttatata agctttcttc 8340

aaataaattt aaaattagat taccttcttt ggaaaaagaa tttcctaaat ttttaagaat 8400

tttcaaagtt ttacatatta gtttttagaa cctaatccgt tttaaaattg tactatgaga 8460

aagctttttt ttgaaagttg taaagcatta atacaaataa tacaaatata attattacca 8520

tcacattcca gagaatatgg ctttttctaa actttcaatt tagaaaacat acattaaggg 8580

agaatctctg ccctcctttt cagctctgaa gatcagcttt tctactcaga cacatgcaca 8640

caccccttcc aagtgtcatg tttatgggaa catttgggaa atgttttcca gatgttttat 8700

tttttccctt ttatagtttg ttgacattta attttactta aagatgacaa ttttaatcgg 8760

aaatgttaga ggtacaacat agtgaggttc tagctagctt tatacttttg aaaaatattt 8820

ttgtttctac tgctttttac aagtactagt cctctcagtg atactggtgg tgttcagtat 8880

gaatccatag aaagaaaaca aaatttgttg tttaaaaaaa gcagagtaat gaatgaattt 8940

cagttttgaa aacaacataa tttgaaaaca ctgttatact aacatggcaa ggtgttaatt 9000

aaatataaga gtaaggtagt aagttctttt agagcacctg tttaaattta ctccagtaat 9060

catcttaagg attgatagtc accatcactt attggcttaa aagttatatt tcatggaata 9120

ttatcagtgt taaatccaag ctttgtggag ctttaagtga tggtggtgaa aaagttggtg 9180

tttatgagag agtggtgggg tgtctagtca ttagtgaagt taaacatcaa cctgttttag 9240

aaagaatttt ttagtcttgc ctaaagtaaa ccagaagtgt ctagtgttta aatctttatt 9300

tagaatgctt ctcttaaaag tattttttgt tttgggtagt attaaataat cagtaaataa 9360

tctatttcag tagtaaataa tgaattaaga tgatgatgaa tgaggattaa cacactggtc 9420

tggagactgg ggttttattt cagtgggtta gctgtgtgtg acatgttggg caattactca 9480

gctgttttaa cagcttccag atatgcagta tggtgcctgt actactcaaa agttgatttt 9540

ggtttaattc atctttaagg tacctcccag ctctaaaact atgattctag gctgtgtaat 9600

ggggttattc ctactttatt ctctttcctt ttttaagggt tcattttata cttaataagc 9660

atccatttct tgggtcacct acagtctttg ttctcctaag gattaaaata gaaaattcat 9720

acataacaag caaatgatga cattttccta aatgctcctt attggttaac cactgaatat 9780

atgaacacat atgaatattg tcattcatgt acttaaattc atttagcaaa ctatttgaac 9840

acttacatgt gcagtgtttg gtgaacatga catgaggaac tagtagtaag taaaatcttc 9900

cccccaaaat tcattgtggc ttaaataaat atgaacataa tcattactac ttaatatact 9960

gagagggaat cttaataaac ttggaactgg gagggaatat ttgtatacat tgggtaaagg 10020

gttaggctag atgacatcta aggggtctga gtgaatcata tcataatttt tataacacat 10080

ttcacatact aaacatcagt tggccccata cctgattaag ttacaaaatt taggagactt 10140

aacattaagg acttacaggt tgagacagcc cgtatttcac aacattattt tgacacttga 10200

ctctattcca gagttgttgc tatacaaggc atgtggcaga acaaaaaaaa agctggtgtt 10260

gatataagag ctttttaccc agtattgaca gtgagcaact ttctttcttt tttttttttt 10320

ttcttttttt tttttttgag atgggttcgc tctgttgccc aggctggtgt gcagtggtgc 10380

gatctcagct cactgcaacc tccacctccc gggttgaagc gattgtcttg cctcagcctt 10440

ccaagtagct ggaattacag gtgcccgccg ccacacctgg ctaatttttg tatttttagt 10500

agagacgggg cttcaccatg ttggccaggc tagtctcgaa ctcttgacct caagtgatcc 10560

acctgccttg gcctccctaa gtgctgggat tacaggcatg agccaccaca cctgtccgac 10620

agtgtagcaa ctttctaaaa ctgaaaaatc tcaaaggaga tcattggaac tgacttgttc 10680

atttattttt tgtttttaaa ttaagaaaga ttacacaaaa taagtgttac tgtactttaa 10740

gctattacaa atatccaact tttaaagata tgtaagaatc agtaatattc tagaaagcac 10800

atatatagta aaagggcatc ctttaaatgt agaacgggta aacatgaaac agttccatgc 10860

ttgaattgtt aagtatctag ggggtaaaca ttgaatggga gaatcattta ttgggttaag 10920

gtcccttcct tgtcattctg ggatctgtga atcacattgt aattcctgtt gacaaagctt 10980

tacttgttaa catcagttga tactgacatt ctccataaag atatagaatg aaaatatcta 11040

ttaaaaatag tttatcattg ttttagcttt tttgttttgt ttgttttgag acagagtctc 11100

actgtcaccc aggcttgagt gcagcggtgt gatcttggct caatgcaacc tccacctccc 11160

aggttcaata gattctccca ccttggcctc ccaagtagct gggattactg gcatgcacca 11220

ctatgcctgg ccagtttttt gtatttttag tagagatggg gtttcaccat gttggccagg 11280

ctggactcga actcctgacc tcaagtgatc cgcccacctc agcctcccaa agtgccggga 11340

ttataggcat gagccactgc gcctagcctg ttgcagcttt ttaaagcagg aaaatatcca 11400

tataaactgt tgggttagaa tctatattag aatctttcaa actaattgaa aacaggaaga 11460

ctatcatcta agtagccaga taatctgggt ttcaaaaagt tattccatgg tactggttta 11520

aaaaatactt ttcaagtgtt ttaattttta aagtgtaact aattcttcaa atatgttatg 11580

ctgttaaaat atgtattcca taagtacttt ttgtatatgt attcttaaat tttaaaaagt 11640

caactgaatg cgcaaagatg atataatttt ggatgtagac atttaaacta gattcccagt 11700

cctctccttc aaaagcttgg tctttgtttt tcctataggg aaaaaagtca aaataagttc 11760

caaaaactat cctcaaagta gtattgtgct tgtagtaaat gaaggttgga tggatggata 11820

ctgacaatgg tggcaggcat ttcaagcctt ttaaattagt actttttgtc gtcttgctta 11880

ttaaaatttt gttaatttta gcaaagacca attgttgtga taaactggtg ttttttggat 11940

gcttcaagca cacgttaacc aattttttaa ttcccctttt ggttcctccc attgttctaa 12000

aataggactt tcatattatt aaaacctcaa aagatgatcc acccaggatg aacaaagatc 12060

accaagggga aagaaaacat tttttatctt tacagaaaac atgttaagat tatatataga 12120

tgtattcttt acattggata ttgtattaga gtcctcctta caagaaatga aatagttttt 12180

agcactctta gcattagagt tcctagattg gtgttgatag ctacagtttt aaaatgtata 12240

acctgaaaat gaaggttaat tttgcattgt aagagcacat ttgatctatg taaaaagtgt 12300

ccatttggtg tattttttta aaaaagagaa agcactttca tattaagtag catgtgtatg 12360

aatttagatt ttcatatttg ttgtgtctgt attcagtgaa gtaaattgag catttaaatg 12420

tttgttgatg gcaacattaa ctattaaatt aaagcacctt atactctgct gcttaacttg 12480

cttgtaattg cacctttgtt acctgcacat tttcatatag aatattgttg taacattgct 12540

tcatgtgggt ctggatggaa gattagtggg cctacaggat catttattta tattgtttat 12600

attacaataa tatattgtag atcagttgta agttcatttc tttacaaata aaagcctctt 12660

ccatttgact ggaaaaaaaa aaaaaaa 12687

<210> 5

<211> 5368

<212> DNA

<213> genus, species, homo sapiens (human)

<400> 5

gtttggcttc acggaaccct gtacgcatgc tcctacgctg aactttagga gccagtctaa 60

ggcctaggcg cagacgcact gagcctaagc agccggtgat ggcggcagcg gctgtggtgg 120

ctgcggcggg tccgggccca tgaggcgacg aaggaggcgg gacggctttt acccagcccc 180

ggacttccga gacagggaag ctgaggacat ggcaggagtg tttgacatag acctggacca 240

gccagaggac gcgggctctg aggatgagct ggaggagggg ggtcagttaa atgaaagcat 300

ggaccatggg ggagttggac catatgaact tggcatggaa cattgtgaga aatttgaaat 360

ctcagaaact agtgtgaaca gagggccaga aaaaatcaga ccagaatgtt ttgagctact 420

tcgggtactt ggtaaagggg gctatggaaa ggtttttcaa gtacgaaaag taacaggagc 480

aaatactggg aaaatatttg ccatgaaggt gcttaaaaag gcaatgatag taagaaatgc 540

taaagataca gctcatacaa aagcagaacg gaatattctg gaggaagtaa agcatccctt 600

catcgtggat ttaatttatg cctttcagac tggtggaaaa ctctacctca tccttgagta 660

tctcagtgga ggagaactat ttatgcagtt agaaagagag ggaatattta tggaagacac 720

tgcctgcttt tacttggcag aaatctccat ggctttgggg catttacatc aaaaggggat 780

catctacaga gacctgaagc cggagaatat catgcttaat caccaaggtc atgtgaaact 840

aacagacttt ggactatgca aagaatctat tcatgatgga acagtcacac acacattttg 900

tggaacaata gaatacatgg cccctgaaat cttgatgaga agtggccaca atcgtgctgt 960

ggattggtgg agtttgggag cattaatgta tgacatgctg actggagcac ccccattcac 1020

tggggagaat agaaagaaaa caattgacaa aatcctcaaa tgtaaactca atttgcctcc 1080

ctacctcaca caagaagcca gagatctgct taaaaagctg ctgaaaagaa atgctgcttc 1140

tcgtctggga gctggtcctg gggacgctgg agaagttcaa gctcatccat tctttagaca 1200

cattaactgg gaagaacttc tggctcgaaa ggtggagccc ccctttaaac ctctgttgca 1260

atctgaagag gatgtaagtc agtttgattc caagtttaca cgtcagacac ctgtcgacag 1320

cccagatgac tcaactctca gtgaaagtgc caatcaggtc tttctgggtt ttacatatgt 1380

ggctccatct gtacttgaaa gtgtgaaaga aaagttttcc tttgaaccaa aaatccgatc 1440

acctcgaaga tttattggca gcccacgaac acctgtcagc ccagtcaaat tttctcctgg 1500

ggatttctgg ggaagaggtg cttcggccag cacagcaaat cctcagacac ctgtggaata 1560

cccaatggaa acaagtggca tagagcagat ggatgtgaca atgagtgggg aagcatcggc 1620

accacttcca atacgacagc cgaactctgg gccatacaaa aaacaagctt ttcccatgat 1680

ctccaaacgg ccagagcacc tgcgtatgaa tctatgacag agcaatgctt ttaatgaatt 1740

taaggcaaaa aaggtggaga gggagatgtg tgagcatcct gcaaggtgaa acgactcaaa 1800

atgacagttt cagagagtca atgtcattac atagaacact tcagacacag gaaaaataaa 1860

cgtggatttt aaaaaatcaa tcaatggtgc aaaaaaaaac ttaaagcaaa atagtattgc 1920

tgaactctta ggcacatcaa ttaattgatt cctcgcgaca tcttctcaac cttatcaagg 1980

attttcatgt tgatgactcg aaactgacag tattaagggt aggatgttgc ttctgaatca 2040

ctgttgagtt ctgattgtgt tgaagaaggg ttatcctttc attaggcaaa gtacaaaatt 2100

gcctataata cttgcaacta aggacaaatt agcatgcaag cttggtcaaa ctttttccag 2160

caaaatggaa gcaaagacaa aagaaactta ccaattgatg ttttacgtgc aaacaacctg 2220

aatctttttt ttatataaat atatattttt caaatagatt tttgattcag ctcattatga 2280

aaaacatccc aaactttaaa atgcgaaatt attggttggt gtgaagaaag ccagacaact 2340

tctgtttctt ctcttggtga aataataaaa tgcaaatgaa tcattgttaa ccacagctgt 2400

ggctcgtttg agggattggg gtggacctgg ggtttatttt cagtaaccca gctgcaatac 2460

ctgtctgtaa tatgagaaaa aaaaaatgaa tctatttaat catttctact tgcagtactg 2520

ctatgtgcta agcttaactg gaagccttgg aatgggcata agttgtatgt cctacatttc 2580

atcattgtcc cgggcctgca ttgcactgga aaaaaaaatc gccacctgtt cttacaccag 2640

tatttggttc aagacaccaa atgtcttcag cccatggctg aagaacaaca gaagagagtc 2700

aggataaaaa atacatactg tggtcggcaa ggtgagggag atagggatat ccaggggaag 2760

agggtgttgc tgtggcccac tctctgtcta atctctttac agcaaattgg taagattttc 2820

agttttactt ctttctactg tttctgctgt ctaccttcct tatatttttt tcctcaacag 2880

ttttaaaaag aaaaaaaggt ctattttttt ttctcctata cttgggctac attttttgat 2940

tgtaaaaata tttgatggcc ttttgatgaa tgtcttccac agtaaagaaa acttagtggc 3000

ttaatttagg aaacatgtta acaggacact atgtttttga aattgtaaca aaatctacat 3060

aaatgattta caggttaaaa gaataaaaat aaaggtaact ttacctttct taaatatttc 3120

ctgccttaaa gagagcattt ccatgacttt agctggtgaa agggtttaat atctgcagag 3180

ctttataaaa atatatttca gtgcatactg gtataataga tgatcatgca gttgcagttg 3240

agttgtatca ccttttttgt ttgtctttta taatgtcttc agtctgagtg tgcaaagtca 3300

atttgtaata ttttgcaacc ctaggatttt tttaaataga tgctgcttgc tatgttttca 3360

aacctttttg agccatagga tccaagccat aaaattcttt atgcatgttg aattcagtca 3420

gaaaagagca aggctttgct ttttgaaatt gcaactcaaa tgagatggga tgaaatccta 3480

tgacagtaag caaaaacaga accatgaaaa atgattggac atacaccttt tcaattgtgg 3540

caataattga aagaatcgat aaaagttcat ctttggacag aaagccttta aaaaaaaaat 3600

cactccctct tccccctcct cccttattgc agcagcctac tgagaacttt gactgttgct 3660

ggtaaattag aagctacaat aataattaag ggcagaaatt atacttaaaa agtgcagatc 3720

cttgttcttt gacaatttgt gatgtctgaa aaaacagaac ccgaaaagct atggtgatat 3780

gtacaggcat tatttcagac tgtaaatggc ttgtgatact cttgatactt gttttcaaat 3840

atgtttacta actgtagtgt tgactgcctg accaaattcc agtgaaactt atacaccaaa 3900

atattcttcc taggtcctat ttgctagtaa catgagcact gtgattggct ggctataacc 3960

accccagtta aaccattttc ataattagta gtgccagcaa tagtggcaaa cactgcaact 4020

tttctgcata aaaagcatta attgcacagc taccatccac acaaatacat agtttttctg 4080

acttcacatt tattaagtga aatttatttc ccatgctgtg gaaagtttat tgagaacttg 4140

tttcataaat ggatatccct actatgactg tgaaaacatg tcaagtgtca cattagtgtc 4200

acagacagaa agcacacacc tatgcaatat ggcttatcta tatttatttg taaaaatcca 4260

agcatagttt aaaatatgat gtcgatatta ctagtcttga gtttctaaga gggttcttta 4320

tgttatacca ggtaagtgta taaaagagat taagtgcttt tttttcatca cttgattatt 4380

ttctttaaaa tcagctatta caggatattt ttttatttta tacatgctgt tttttaatta 4440

aaatataatc actgaagttt actaatttga ttttataagg tttgtagcat tacagaataa 4500

ctaaactggg atttataaac cagctgtgat taacaatgta aagtattaat tattgaactt 4560

tgaaccagat ttttaggaaa attatgttct ttttccccct ttatggtctt aactaatttg 4620

aatccttcaa gaaggatttt tccatactat tttttaagat agaagataat ttgtgggcag 4680

gggtggagga tgcatgtatg atactccata aattcaacat tctttactat aggtaatgaa 4740

tgattataaa caagatgcat cttagatagt attaatatac tgagccttgg attatatatt 4800

taatatagga cctattttga atattcagtt aatcatatgg ttcctagctt acaagggcta 4860

gatctaagat tattcccatg agaaatgttg aatttatgaa gaatagattt taaggctttg 4920

aaaatggtta atttctcaaa aacatcaatg tccaaacatc tacctttttt cataggagta 4980

gacactagca agctggacaa actatcacaa aagtatttgt cacacataac ctgtggtctg 5040

ttgctgatta atacagtact ttttcttgtg tgattcttaa cattatagca caagtattat 5100

ctcagtggat tatccggaat aacatctgaa agatgggttc atctatgttt gtgtttgctc 5160

tttaaactat tgtttctcct atcccaagtt cgctttgcat ctatcagtaa ataaaattct 5220

tcagctgcct tattaggagt gctatgaggg taacacctgt tctgcttttc atcttgtatt 5280

tagttgactg tattatttga tttcggattg aatgaatgta aatagaaatt aaatgcaaat 5340

ttgaatgaac ataaaaaaaa aaaaaaaa 5368

<210> 6

<211> 4270

<212> DNA

<213> genus, species, Homo sapiens (human)

<400> 6

gagggccgct gtcactcagc cccgcgggcc aatagaaaag gggtgaaccc cgccttcttc 60

ctgagttgtg ctgcgggcat gcgcactggg cgtccccacg ccaccgccca tcagctgaga 120

attgcagctg agggctccgg ggtaggtggg tgacggcggt cggaggtgta ggagggagcc 180

gtggaggtcc aggtgactgc ttagaaaact gcacagcatc tgatgaaatt agcgaataag 240

aacatcaacc atgtcttaca ctccaggagt tggtggtgac cccgcccagt tggcccagag 300

gatctcttct aacatccaga agatcacaca gtgttctgtg gaaatacaaa gaactctgaa 360

tcaacttgga acacctcaag attcacctga attgaggcaa cagttgcaac agaagcagca 420

gtatactaac cagcttgcca aagaaacaga taagtacatt aaagagtttg gatctctgcc 480

caccaccccc agtgaacagc gtcaaaggaa aatacagaag gatcgcttag tggcagagtt 540

cacaacatca ctgacaaact tccagaaggt ccagaggcag gctgctgagc gagagaaaga 600

gtttgttgct cgagtaagag ccagttccag agtgtctggc agttttcctg aggacagctc 660

aaaagaaagg aatcttgtat cctgggaaag ccaaactcaa cctcaagtgc aggtgcagga 720

tgaagaaatt acagaggatg acctccgtct tattcatgag agagaatctt ctatcaggca 780

acttgaagct gatattatgg atattaatga aatatttaaa gatttgggaa tgatgattca 840

tgaacaagga gatgtaatag atagcataga agccaatgtg gaaaatgcag aggtgcacgt 900

tcagcaagca aatcagcagc tgtcaagggc agcagattat cagcgcaaat ccagaaaaac 960

cctgtgcatc atcattctta tccttgtcat tggagttgcg attatcagtc tcatcatatg 1020

gggattgaac cactgaagtt ataaaggagc acactgtcgc actacattgt ctaaattatg 1080

taggaagatt cctgtaatca tgttttttta attattattt taaagctatt gtataaagga 1140

tggttcccat actttgttat ttttattggg ggggttgggg tggttccttt ggattaaatc 1200

tgatattttc taatactgaa agattttcta aatgtcactg ctgacataac tcccttggtc 1260

ttcaatttaa tagttgttaa gtttttgccc acattgcata tgcctttcat ttataattta 1320

tttaccctgc ttgacttagt tttgggaatt cgtaaattta aaggtgtgtg tattctgttt 1380

gcatctccct gtcactgtga cacacctaga tgtgtgttac ttcaattaaa attctcaaat 1440

ttaattttga tttgcttcag cagggaaaat attctcaata atgtaaaata attaaggtct 1500

atacatgggt tgtatttttc tggttcacaa cagcacaaag tgtctttcat ttttttgttg 1560

gttttctttt aagatctttt ttaccctgaa gtcggtgaat acttttctag tttatttgat 1620

actctttctg tgtatatatt aagcttttgc tgtagattgc ctagtaaaat tactaaggat 1680

aggttgtttt tacatatggt ctatttaagt ctgatgttta cgggggaaag tgtagttact 1740

aaaaatgttt aacataattt ggaagaagag tatgaacaac caataccaat acctattgcg 1800

tttggattct taagacccca gtttgttatt ccactaaact agttatctta accatatcat 1860

ctggttttgt gggccattat ttaccttccc ttatgtctta tagaataatg gttaatattt 1920

ttaggtcaaa attacttttg gaaagtaact ttcccacaat taactgtttt tgagcacctg 1980

acaaaattta gtgtttacct tgcgtgccat tttgtgtcat ccttcattaa aaaagcaatt 2040

ggaggtttgc cagttatctc acttcccttt ttaaatcaat gttgttttaa tgcactaatc 2100

tgaattctgt aaagaggatt atcttagttt atactttgta ttttataatg ttcttgtata 2160

gcagctcggt actgaaggcg gtgtttaact tggcaagctc tgagacttca aatgggaaca 2220

aatagtaagt agctaagtaa accacatctt tgcaaccaaa ataaagatga gttaaaaggt 2280

atctggttag gcctatttca tgaggactat gctctggtgg gaccacaggt cacctgatac 2340

ttagtgctgt gctgcctgaa acctcagcat ggagacatca ccacatgcac tgtggccatt 2400

cagcatcttt ctggagcacc agttcactcc aggtttttat tttagggtgt catcgatttt 2460

acctactttg tcagactggt agaagttgct ttgcatatca gaaaaactcc atttttttcc 2520

acaaaaggga ttacagaaaa ctcttttgtg agtgagtgat tggaacttag agactcctgt 2580

tgccagaatc agactgccct agaacagaat ggacaatgca gggaggagaa ttcacacaaa 2640

cagcacctgt tctgaggcct gtgccagccc accaggcctg ctcaaatgtg gtctttactt 2700

caagtgcaca gaggcacatg aggtttctgg tgataaacca gcgtcttacc gctgttttaa 2760

agtcccatcc ccatggcttt cacaatcagt tccgtttttt ttgctgtact tgataaaatg 2820

tttattctca tacaggtcaa gtacatttac ttctattcac agtgagtacc caataacaac 2880

aaaagcgctt acaaatttgg ggggcgtgat tttagtacct ttatttgaag tgtaatcatt 2940

ttaaattatt attattttaa ctggggcagt tatcagtggt ttaaacagga actttagtgg 3000

cttcaatttg tttaagaaac atattaagtt tgagggaaaa atttcccatg aaatatttgg 3060

aacgtaagag tagtattgat tagagaaaat taaataagaa acatagtatg gtagccaaat 3120

tttttaaaaa atcttgaact tttctgtagg tcagttttag aactgctgtg aaaagtgaag 3180

gttgccctgt ggagattaaa attagagttg ttttcataac tgacagcatg gtgaatccat 3240

ttgagtcaaa gtgaagaatt tcctcatcaa gtgactatac atttgttttt gtgtgctcaa 3300

aagaaatact caaacacaga ctgatattaa ccagccaggt aaattgaacg acaatgtggc 3360

attaggtatt tggctgttta ttggtcgtta aatactatgg ttttgcaata tgattgatgg 3420

taaaagagtg atgtcatatt gatactagag tagcttgttt ttttagtagg tgtgggacca 3480

tctcttttac aagtgcaact cagtctagga cagccatgga tgcagtgtct ggagttggac 3540

cctctgagcc cgctggctgc cccagtagca tctgcattgg tgaccaagga cactgcactt 3600

tgaagaggtc gccactgggt tatttagtgt cttcactgct tttgttaaaa attgtaaaat 3660

tttgtacaca aaaagttgtg tttttgaata tcaattgttt agacacacct acaatgataa 3720

ataagtgcct ttaaaggccc ctctttccat gaaatacatc tgtggtttag caaggaaagt 3780

acaaaatagt ttatgtagtt ggtataattt ttatttgtgt cttcatgtag aaaaaatgaa 3840

tgtcataata aaatataaaa cttacgtaaa gaaaataaag tcattgtcca ccttaatagc 3900

taaggtccac aagggtaact tatgcagcat ttattttttt tgaaagtcaa aattgaattt 3960

atttctttca catggctggt ttgctgcaat atgaagtttc agaatgggct gaagtaagtt 4020

gattgaggga tttgagttga atgacatttt caagttcatt taaatatgat aaaaattcat 4080

tggtggtaaa taacatctgt ctttcctgga aaaaaaaaag ttgtgtattt tcatgattca 4140

gttaaaacaa aaaatgagcc tgtgaatccc aggccttttt agtcctccat aacatttgaa 4200

cagtttgact tgtcagcaaa gaaatacact tatcaaattt taaaccaatg ggagcctgaa 4260

agtgttacag 4270

<210> 7

<211> 21

<212> DNA

<213> Artificial sequence

<400> 7

ggatcttgtc agtgacgttg t 21

<210> 8

<211> 19

<212> DNA

<213> Artificial sequence

<400> 8

gtagcgattc cggcactgt 19

<210> 9

<211> 20

<212> DNA

<213> Artificial sequence

<400> 9

acctgaggca gctcttttgc 20

<210> 10

<211> 25

<212> DNA

<213> Artificial sequence

<400> 10

ggatgtgtgt gctccttcta aagaa 25

<210> 11

<211> 23

<212> DNA

<213> Artificial sequence

<400> 11

tttctctcaa aggactacga gat 23

<210> 12

<211> 18

<212> DNA

<213> Artificial sequence

<400> 12

agcagccctc aggtcaaa 18

<210> 13

<211> 20

<212> DNA

<213> Artificial sequence

<400> 13

cagttctacc agcggcagta 20

<210> 14

<211> 20

<212> DNA

<213> Artificial sequence

<400> 14

ggtagcgggt gtgaaaatca 20

<210> 15

<211> 19

<212> DNA

<213> Artificial sequence

<400> 15

ccgaactctg ggccataca 19

<210> 16

<211> 22

<212> DNA

<213> Artificial sequence

<400> 16

ttgcaggatg ctcacacatc tc 22

<210> 17

<211> 25

<212> DNA

<213> Artificial sequence

<400> 17

ctcttttgtg agtgagtgat tggaa 25

<210> 18

<211> 21

<212> DNA

<213> Artificial sequence

<400> 18

ccctgcattg tccattctgt t 21

<210> 19

<211> 26

<212> DNA

<213> Artificial sequence

<400> 19

aactcctgta gccgaatcta ccgctc 26

<210> 20

<211> 22

<212> DNA

<213> Artificial sequence

<400> 20

tagccacccc caccccactt gc 22

<210> 21

<211> 28

<212> DNA

<213> Artificial sequence

<400> 21

tcaaaatcct ctggcctctc ctacgaac 28

<210> 22

<211> 22

<212> DNA

<213> Artificial sequence

<400> 22

cccctaccct aacggcaagc ca 22

<210> 23

<211> 22

<212> DNA

<213> Artificial sequence

<400> 23

caaacggcca gagcacctgc gt 22

<210> 24

<211> 29

<212> DNA

<213> Artificial sequence

<400> 24

actcctgttg ccagaatcag actgcccta 29

<210> 25

<211> 24

<212> DNA

<213> Artificial sequence

<400> 25

accatgagaa gtatgacaac agcc 24

<210> 26

<211> 23

<212> DNA

<213> Artificial sequence

<400> 26

cacgatacca aagttgtcat gga 23

<210> 27

<211> 23

<212> DNA

<213> Artificial sequence

<400> 27

tcagcaatgc ctcctgcacc acc 23

<210> 28

<211> 1421

<212> DNA

<213> genus, species, Homo sapiens (human)

<400> 28

gcctcaagac cttgggctgg gactggctga gcctggcggg aggcggggtc cgagtcaccg 60

cctgccgccg cgcccccggt ttctataaat tgagcccgca gcctcccgct tcgctctctg 120

ctcctcctgt tcgacagtca gccgcatctt cttttgcgtc gccagccgag ccacatcgct 180

cagacaccat ggggaaggtg aaggtcggag tcaacggatt tggtcgtatt gggcgcctgg 240

tcaccagggc tgcttttaac tctggtaaag tggatattgt tgccatcaat gaccccttca 300

ttgacctcaa ctacatggtt tacatgttcc aatatgattc cacccatggc aaattccatg 360

gcaccgtcaa ggctgagaac gggaagcttg tcatcaatgg aaatcccatc accatcttcc 420

aggagcgaga tccctccaaa atcaagtggg gcgatgctgg cgctgagtac gtcgtggagt 480

ccactggcgt cttcaccacc atggagaagg ctggggctca tttgcagggg ggagccaaaa 540

gggtcatcat ctctgccccc tctgctgatg cccccatgtt cgtcatgggt gtgaaccatg 600

agaagtatga caacagcctc aagatcatca gcaatgcctc ctgcaccacc aactgcttag 660

cacccctggc caaggtcatc catgacaact ttggtatcgt ggaaggactc atgaccacag 720

tccatgccat cactgccacc cagaagactg tggatggccc ctccgggaaa ctgtggcgtg 780

atggccgcgg ggctctccag aacatcatcc ctgcctctac tggcgctgcc aaggctgtgg 840

gcaaggtcat ccctgagctg aacgggaagc tcactggcat ggccttccgt gtccccactg 900

ccaacgtgtc agtggtggac ctgacctgcc gtctagaaaa acctgccaaa tatgatgaca 960

tcaagaaggt ggtgaagcag gcgtcggagg gccccctcaa gggcatcctg ggctacactg 1020

agcaccaggt ggtctcctct gacttcaaca gcgacaccca ctcctccacc tttgacgctg 1080

gggctggcat tgccctcaac gaccactttg tcaagctcat ttcctggtat gacaacgaat 1140

ttggctacag caacagggtg gtggacctca tggcccacat ggcctccaag gagtaagacc 1200

cctggaccac cagccccagc aagagcacaa gaggaagaga gagaccctca ctgctgggga 1260

gtccctgcca cactcagtcc cccaccacac tgaatctccc ctcctcacag ttgccatgta 1320

gaccccttga agaggggagg ggcctaggga gccgcacctt gtcatgtacc atcaataaag 1380

taccctgtgc tcaaccagtt aaaaaaaaaa aaaaaaaaaa a 1421

Claims (7)

1. A group of liver cancer gene markers is characterized in that: consists of an EP400 gene, an MAPK1IP1L gene, a NUFIP2 gene, a PHC3 gene, an RPS6KB1 gene and an STX7 gene.

2. The panel of liver cancer gene markers of claim 1, wherein: the EP400 gene is a polynucleotide sequence shown in a sequence table SEQ ID NO. 1, the MAPK1IP1L gene is a polynucleotide sequence shown in a sequence table SEQ ID NO. 2, the NUFIP2 gene is a polynucleotide sequence shown in a sequence table SEQ ID NO. 3, the PHC3 gene is a polynucleotide sequence shown in a sequence table SEQ ID NO. 4, the RPS6KB1 gene is a polynucleotide sequence shown in a sequence table SEQ ID NO. 5, and the STX7 gene is a polynucleotide sequence shown in a sequence table SEQ ID NO. 6.

3. Use of the reagent for detecting the expression level of the liver cancer gene marker according to claim 1 or 2 in the preparation of a product for detecting liver cancer.

4. Use according to claim 3, characterized in that: the product for detecting liver cancer is a real-time quantitative PCR kit, an RNA sequencing kit or a gene chip.

5. Use according to claim 3, characterized in that: the reagent for detecting the liver cancer gene marker is a primer and/or a probe for detecting the liver cancer gene marker.

6. Use according to claim 5, characterized in that: the primer is a primer for specifically amplifying SEQ ID NO:1 to SEQ ID NO:6 gene sequence primer; the probe is a probe which can be matched with SEQ ID NO:1 to SEQ ID NO: 6.

7. Use according to claim 6, characterized in that: the primer is a sequence shown in SEQ ID NO 7-18 of a sequence table; the probe is a sequence shown in a sequence table SEQ ID NO. 19-SEQ ID NO. 24.

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