KR101802407B1 - Expression validation of protein expressed uniquely by scrapie prion protein and use for development of biomarker for the diagnosis of Creutzfeldt-Jakob disease - Google Patents

Expression validation of protein expressed uniquely by scrapie prion protein and use for development of biomarker for the diagnosis of Creutzfeldt-Jakob disease Download PDF

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KR101802407B1
KR101802407B1 KR1020150052375A KR20150052375A KR101802407B1 KR 101802407 B1 KR101802407 B1 KR 101802407B1 KR 1020150052375 A KR1020150052375 A KR 1020150052375A KR 20150052375 A KR20150052375 A KR 20150052375A KR 101802407 B1 KR101802407 B1 KR 101802407B1
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protein
acsbg1
creutzfeldt
jakob disease
kit
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KR20160122886A (en
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이영선
주영란
김수연
김지경
최보영
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대한민국
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Abstract

The present invention relates to the verification of specific protein expression in brain tissue of transformed prion-infected mice. Identification of information and biological pathways of whole proteins identified in transformed prion-infected brain tissues can be used to identify mechanisms of prion in vivo and to identify additional candidate diagnostic markers It is worth using as a selectable database. Blood fluid, such as blood, circulates in almost all tissues of the body and is secreted by specific tissues or contains proteins from the dead cells of the tissues, reflecting the state of the organ, and can also be easily collected for non-invasive ) Can be diagnosed. The present invention has confirmed the expression of ACSBG1 protein by selecting a candidate diagnostic marker which has not been reported in prion disease in a biological pathway analyzed from infected mouse brain tissue protein information. The ACSBG1 protein expression was increased in the transformed prion-infected mouse compared to the normal group, and thus it was useful as a method for screening candidates for Crotsfeldt-Jakob disease diagnosis kit or biochip by using the ACSBG1 protein expression change and a method for screening candidates for treatment of Crotsfeldt- Can be used.

Description

Expression validation of protein expression uniquely by scrimie prion protein and use of a biomarker for diagnosis of Creutzfeldt-Jakob disease.

The present invention relates to a specific protein expression test in brain tissue of a transformed prion-infected mouse, and relates to a method for screening for a protein specific for ACSBG1 (acyl-CoA synthetase bubblegum family member 1) protein or an antibody or an aptamer (Creutzfeldt-Jakob disease) diagnostic kit and a method for screening candidates for treatment of Creutzfeldt-Jakob disease.

Creutzfeldt-Jakob disease (CJD) is one of the transmissible spongiform encephalopathy (TSE) that occurs in humans. It is caused by a scrapie prion protein (PrP Sc ) , But the physiological function of the prion has not yet been elucidated. Normal prions are soluble and are degraded by protein hydrolytic enzymes and have a large number of a-helix structures but insoluble transformed prions with a b-sheet structure And is resistant to proteolytic enzymes. It occurs in 1-2 people per 1 million population and after a very long incubation period ranging from months to years, causes central nervous system abnormalities and causes a neuropathologic lesion in the brain tissue, vacuolation. Inflammatory and disease-specific immune responses are not present at all, and once a condition is manifested, it has a fatal prognosis of 100% death. Dementia, myoclonus, ataxia, characteristic periodic EEG findings, and the like. Approximately 85-90% of human prion diseases are sporadic CJD and hereditary CJD is approximately 10-15%. Variants that are transmitted to humans through ingestion of carcass of animals infected with degenerated prions, blood transfusions, organ transplants, About 1%. CJD is a system in which modified prions (PrP Sc ) enter the body to transform normal prions (PrP c ) into modified prion structures and form oligomers or aggregates between transformed prion molecules, resulting in central nervous system degeneration and neurological symptoms . ≪ / RTI >

Similar diseases include bovine spongiform encephalopathy (BSE) and positive scrapie, and human prion diseases are caused by the culprit, which is spread by consciousness, Kuru) was first announced in the 1950s. CJD has not developed preventive and therapeutic methods until now. Especially, it has been reported that the intake of beef infected with mad cow disease is related to variant CJD (variant CJD, vCJD) Europe and many other countries have established a surveillance system for them and are strengthening research on diagnostic methods and treatment methods. Currently, CJD can be confirmed only by brain autopsy or biopsy. The detection of 14-3-3 protein, which is used as a diagnostic indicator, has a low specificity and is limited in detection of other brain diseases such as Alzheimer. In addition, in the case of vCJD, only about 50% of the samples are positive for 14-3-3 protein. Therefore, it is necessary to find a diagnostic indicator with high sensitivity and specificity for the early diagnosis of prion disease.

Acquisition of the National Institutes of Health by the Center for Disease Control In the CJD diagnosis method performed by the Common Infection Division, 14-3-3 protein detection method and prion gene polymorphism analysis are performed by Western blot using the standard method designated by WHO. 14-3-3 protein has been reported to be expressed in two-dimensional electrophoresis (2-DE) from cerebrospinal fluid of CJD patients and has been used as a diagnostic indicator for CJD until now. The biomarkers found in 2-DE include cystatin C, H-FABP family, and Apo E (apolipoprotein E). Recently, transferrin, neuron-specific enol (Neuron-specific enolase) and urinary α1-antichymotrypsin (α1-ACT) have been proposed as new biomarkers for CJD and have been verified by various experiments.

Acyl-CoA synthetase is a protein that plays an important role in metabolism and myelinogenesis of very long chain fatty acids (VLCFA). As an energy producer, it affects the mitochondrial matrix as respiration and as an electron donor. ACSBG1 (acyl-CoA synthetase bubblegum family member 1) is known to interact with 14-3-3 beta used for laboratory diagnosis. It is expressed intensely in the brain, adrenal gland and testis and is an important enzyme in the early stages of β-oxidation of VLCFA. ACSBG1 has been reported to affect neurodegenerative diseases such as X-linked adrenoleukodystrophy (X-ALD) and is known to be associated with the accumulation of VLCFA. It is known as Lorenzo oil disease, and the concentration of saturated fatty acid increases continuously in the blood and accumulates in the living body. In particular, the accumulation of saturated fatty acids in the myelin surrounding the nerve cell surface dissolves myelin and results in loss of function of the nerve cells.

In addition to the above biomarkers, the present inventors predicted that the increased expression of ACSBG1 in the brain tissue would affect the function of nerve cells, and thus a transformed prion-infected mouse was produced. Identification of information and biological pathways of whole proteins identified in transformed prion-infected brain tissue is valuable as a database to identify mechanisms of prion in vivo mechanisms and to select additional candidate diagnostic markers. Blood fluid, such as blood, circulates in almost all tissues of the body and is secreted by specific tissues or contains proteins from the dead cells of the tissues, reflecting the state of the organ, and can also be easily collected for non-invasive ) Can be diagnosed. The present invention has confirmed the expression of ACSBG1 protein by selecting a candidate diagnostic marker which has not been reported in prion disease in a biological pathway analyzed from infected mouse brain tissue protein information. The expression of the ACSBG1 protein can be usefully used as a kit for the diagnosis of Creutzfeldt-Jakob disease or a biochip for screening candidate substances for treatment of Creutzfeldt-Jakob disease using the same To complete the present invention.

An object of the present invention is to provide a kit for the diagnosis of Creutzfeldt-Jakob disease comprising an antibody or an aptamer which specifically binds to ACSBG1 (acyl-CoA synthetase bubblegum family member 1) - < / RTI >

In order to achieve the above object, the present invention provides a kit for the diagnosis of Creutzfeldt-Jakob disease comprising an antibody or an aptamer specifically binding to an acyl-CoA synthetase bubblegum family member 1 (ACSBG1) Lt; / RTI >

The present invention also provides a biochip for the diagnosis of Creutzfeldt-Jakob disease comprising an acyl-CoA synthetase bubblegum family member 1 (ACSBG1) antibody or an aptamer.

The present invention also provides a kit for the diagnosis of Creutzfeldt-Jakob disease comprising a pair of primers that specifically amplify an ACSBG1 (acyl-CoA synthetase bubblegum family member 1) gene, or an antisense nucleotide complementarily binding to the ACSBG1 gene .

In addition,

1) measuring the expression level of ACSBG1 (acyl-CoA synthetase bubblegum family member 1) protein from the sample isolated from the subject; And

2) a step of selecting an individual having a higher expression level than the normal expression level of the protein of step 1), to provide information on the diagnosis of Creutzfeldt-Jakob disease.

In addition,

1) treating a test compound or composition with an ACSBG1 (acyl-CoA synthetase bubblegum family member 1) protein or a cell expressing the ACSBG1 protein; And

2) screening a candidate compound for treatment of Creutzfeldt-Jakob disease, comprising the step of screening a test compound or a composition that decreases the expression or activity of the ACSBG1 protein of step 1).

The present invention relates to the verification of specific protein expression in brain tissue of transformed prion-infected mice. Identification of information and biological pathways of whole proteins identified in transformed prion-infected brain tissues can be used to identify mechanisms of prion in vivo and to identify additional candidate diagnostic markers It is worth using as a selectable database. Blood fluid, such as blood, circulates in almost all tissues of the body and is secreted by specific tissues or contains proteins from the dead cells of the tissues, reflecting the state of the organ, and can also be easily collected for non-invasive ) Can be diagnosed. The present invention has confirmed the expression of ACSBG1 protein by selecting a candidate diagnostic marker which has not been reported in prion disease in a biological pathway analyzed from infected mouse brain tissue protein information. The ACSBG1 protein expression was increased in the transformed prion-infected mouse compared to the normal group, and thus it was useful as a method for screening candidates for Crotsfeldt-Jakob disease diagnosis kit or biochip by using the ACSBG1 protein expression change and a method for screening candidates for treatment of Crotsfeldt- Can be used.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a diagram showing the infection of a modified prion-infected mouse of the present invention. FIG.
CTRL: normal mouse brain emulsion inoculation mouse (control group);
ME7: ME7 scrapie infected mouse brain emulsion inoculation mouse; And
PK: Proteolytic enzyme.
2A shows a biological pathway of a protein whose expression is increased in a normal group.
FIG. 2B is a view showing a biological pathway of a protein whose expression is increased in the transformed prion infection group.
Figure 3 shows the difference in expression of ACSBG1 in the control and transformed prion-infected mice:
CTRL: normal mouse brain emulsion inoculation mouse (control group); And
ME7 scrapie infected: ME7 Scrapie infected mice Brain emulsion vaccination mice.

Hereinafter, the present invention will be described in detail.

The present invention provides a kit for the diagnosis of Creutzfeldt-Jakob disease comprising an antibody or an aptamer specifically binding to an acyl-CoA synthetase bubblegum family member 1 (ACSBG1) protein.

The present invention also provides a biochip for the diagnosis of Creutzfeldt-Jakob disease comprising an acyl-CoA synthetase bubblegum family member 1 (ACSBG1) antibody or an aptamer.

Preferably, the ACSBG1 is composed of the amino acid sequence of SEQ ID NO: 1, or is encoded by the nucleotide sequence of SEQ ID NO: 2.

The antibody is preferably bound to a solid substrate and preferably further comprises an antibody for detection bound to the protein. The antibody for detection may be any antibody selected from the group consisting of a coloring enzyme, a fluorescent substance, a radioactive isotope and a colloid. But it is not limited thereto.

The fluorescent substance may be any fluorescent substance used in immunoassay, for example, a coumarin-based fluorescent substance (e.g., 7-hydroxycoumarin, 4-methyl-7-hydroxycoumarin, etc.); Xanthene series fluorescent materials (e.g., ROX, TET, Texas Red, fluorescein, tetramethylrhodamine, etc.); Cyanine-based fluorescent materials (e.g., Cy3, Cy5, etc.); Bodipy-based fluorescent materials (e.g., Bodipy FL, Bodipy 530, Bodipy R6G, Bodipy TMR, etc.); (E.g., Alexa Fluor 488, Alexa Fluor 647, Alexa Fluor 568, Alexa Fluor 594, Alexa Fluor 546, Alexa Fluor 350, Alexa Fluor 555, Alexa Fluor 532, etc.); (DyLight 405, DyLight 488, DyLight 549, DyLight 594, DyLight 633, DyLight 649, DyLight 680, DyLight 750, DyLight 800, etc.) But is not limited thereto.

Kit systems for use with the present invention include, but are not limited to, ELISA plates, dip-stick devices, immunochromatography methods and split-split immune assay devices, flow-through devices, and the like. Preferably, a strip-type or device-type diagnostic kit using immunochromatography can be used. In the diagnosis using the immunochromatography method, the antigen contained in the serum of the sample is reacted with a tracer antibody bound to a colloidal gold particle, and then a nitrocellulose membrane It is possible to visually distinguish between positive and negative by binding with a capture antibody immobilized on the inner surface of the micropores during the movement through the micropore of the micropores.

The kit may include, but is not limited to, an enzyme-linked immunoassay (ELISA) or a lateral flow immunographic assay. As described above, when a sandwich immunoassay and a lateral flow immunographic assay are used, an antibody pair that specifically binds to the protein of the present invention is used to prepare an antibody pair, can do.

The kit of the present invention is not particularly limited as long as it is used in a kit. However, for example, if using ELSIA, a solid-phase supporter; An antibody that specifically binds to the protein of the present invention; And an enzyme-linked immunosorbent assay (ELSIA) reaction solution containing an enzyme-labeled antibody solution for reaction with an antigen and a coloring solution showing an enzyme reaction. More specifically, the enzyme-labeled antibody solution may contain 50 to 150 μl per well of the plate at an appropriate concentration of goat anti-mouse Ig-HRP. The coloring solution may be selected from the group consisting of tetramethylbenzidine (TMB) The reaction blocking solution may be selected from the group consisting of 1N HCl or 1N H 2 SO 4 .

The kit for the diagnosis of Creutzfeldt-Jakob disease according to the present invention can be used as a kit for a reverse transcription polymerase chain reaction (RT-PCR) kit, a competitive PCR kit, a real-time PCR kit, an RNase protection assay (RPA) A kit, a Northern blotting kit, or a DNA chip kit. However, if the mRNA or DNA is complementarily bound to the mRNA or DNA, it may be prepared by any known method known to those skilled in the art. have.

In a specific example of the present invention, the present inventors prepared a modified prion-infected mouse and performed western blotting to confirm the infectivity. As a result, the infectivity of the transformed prion-infected mouse was confirmed. Protein hydrolytic enzymes (PK) reduce the size of the amino acid by cutting off the amino acid end, resulting in un-glycosylated, mono-glycosylated, di-glycosylated, (See Fig. 1), proteins were separated from brain tissue of transformed prion-infected mice and classified and analyzed by function. As a result, it was confirmed that a biological pathway (FIG. 2A) of the protein whose expression was increased in the normal group and a biological pathway of the protein whose expression was increased in the infection group (FIG. 2B) , The ACSBG1 protein was selected as a candidate diagnostic marker and the expression of ACSBG1 was confirmed. As a result, at the expected molecular size of ACSBG1 of 80 kDa, ME7 scrapie infected mouse (See Fig. 3).

Therefore, the ACSBG1 protein of the present invention was produced by transforming prion-infected mice, and the proteins were isolated from the brain tissue that confirmed infectivity, and the biological pathways of the identified proteins were identified. The proteins that were not reported in prion diseases were selected The ACSBG1 protein was selected as a candidate diagnostic indicator and the expression of the ACSBG1 protein was confirmed. As a result, the expression of the ACSBG1 protein in the transformed prion-infected mouse was found to be higher than that of the normal group. Thus, And can be usefully used as a kit for detecting a felt-Jakob disease or a biochip.

The present invention also provides a kit for the diagnosis of Creutzfeldt-Jakob disease comprising a pair of primers that specifically amplify an ACSBG1 (acyl-CoA synthetase bubblegum family member 1) gene, or an antisense nucleotide complementarily binding to the ACSBG1 gene .

The ACSBG1 is preferably composed of the nucleotide sequence of SEQ ID NO: 2.

The kit for the diagnosis of Creutzfeldt-Jakob disease according to the present invention can be used as a kit for a reverse transcription polymerase chain reaction (RT-PCR) kit, a competitive PCR kit, a real-time PCR kit, an RNase protection assay (RPA) A kit, a Northern blotting kit, or a DNA chip kit. However, if the mRNA or DNA is complementarily bound to the mRNA or DNA, it may be prepared by any known method known to those skilled in the art. have.

ACSBG1 of the present invention was prepared by isolating proteins from brain tissues which were confirmed to have infectivity by preparing a transformed prion-infected mouse. The protein pathway of the identified proteins was confirmed, and proteins not reported in prion diseases were selected from the analyzed pathways, Was selected as a candidate diagnostic indicator, and the expression of ACSBG1 protein was confirmed. As a result, expression of the ACSBG1 gene was amplified in the transformed prion-infected mouse as compared with the normal group. Thus, a primer pair specifically amplifying the ACSBG1 gene, Complementarily binding antisense nucleotides can be usefully used as a kit for the diagnosis of Creutzfeldt-Jakob disease.

In addition,

1) measuring the expression level of ACSBG1 (acyl-CoA synthetase bubblegum family member 1) protein from the sample isolated from the subject; And

2) a step of selecting an individual having a higher expression level than the normal expression level of the protein of step 1), to provide information on the diagnosis of Creutzfeldt-Jakob disease.

The sample of step 1) is preferably one selected from the group consisting of blood, serum, plasma, cells or tissue, but is not limited thereto.

The expression level of the above step 1) can be determined by Western blotting, Enzyme-linked immunosorbent assay (ELISA), RT-PCR, immunohistochemical staining, immunoprecipitation and immunofluorescence and immunofluorescence. However, the present invention is not limited thereto.

The ACSBG1 protein of the present invention was prepared by isolating proteins from brain tissues in which infectivity was confirmed by preparing a transformed prion-infected mouse. The protein pathways were identified by the identified proteins, and proteins not reported in prion diseases were selected from the analyzed biological pathways. Protein was selected as a candidate diagnostic indicator and the expression of the ACSBG1 protein was confirmed and the expression of the ACSBG1 protein was increased in the transformed prion-infected mouse as compared with that of the normal group, thereby providing a degree of diagnosis of Creutzfeldt-Jakob disease using the ACSBG1 protein Which can be used as a detection method for proteins.

In addition,

1) treating a test compound or composition with an ACSBG1 (acyl-CoA synthetase bubblegum family member 1) protein or a cell expressing the ACSBG1 protein; And

2) screening a candidate compound for treatment of Creutzfeldt-Jakob disease, comprising the step of screening a test compound or a composition that decreases the expression or activity of the ACSBG1 protein of step 1).

The expression or activity of the protein in step 2) is preferably measured by any one method selected from the group consisting of Western blotting, enzyme-immunochemical detection, RT-PCR, immunohistochemical staining, immunoprecipitation and immunofluorescence , But is not limited thereto.

The ACSBG1 protein of the present invention was prepared by isolating proteins from brain tissues in which infectivity was confirmed by preparing a transformed prion-infected mouse. The protein pathways were identified by the identified proteins, and proteins not reported in prion diseases were selected from the analyzed biological pathways. Protein was selected as a candidate diagnostic indicator and the expression of the ACSBG1 protein was confirmed and the expression of the ACSBG1 protein was increased in the transformed prion-infected mouse as compared with that of the normal group, thereby screening candidates for the treatment of Creutzfeldt-Jakob disease Can be usefully used.

Hereinafter, the present invention will be described in detail with reference to examples.

However, the following examples are illustrative of the present invention, but the present invention is not limited thereto.

< Example  1> modified prion (scrapie prion  protein, PrP Sc ) Production of infected mice

20 μl of 10% ME7 scrapie brain homogenate was injected intraperitoneally into a 4-week-old male C57BL / 6 mouse (Sumatoco Bio Korea), and symptoms were continuously observed. After 171 days of behavioral symptoms . Control (CTRL) mice were injected intraperitoneally with 20 μl of normal mouse brain homogenate in mice of the same age and sex, and brain was extracted on day 171. The mouse brain tissue was perfused with 20 ml of cold PBS (phosphate-buffered saline) buffer solution and then weighed to make a brain homogenate. 10% (w / v) of PBS was added and pulverized and stored at -70 ° C. until the experiment.

< Example  2> modified prion ( PrP Sc ) Infection of infected mice

Western blotting, which is a method for confirming the relatively high specificity using the antigen-antibody reaction, was performed to confirm whether the transformed prions produced in Example 1 were infected with the modified prions.

Specifically, RIPA (radioimmunoprecipitation assay) buffer solution was added to the 10% (w / v) brain homogenate obtained in Example 1, and the protein hydrolyzate (proteinase K, 1 mg / ml) And reacted at 37 ° C for 1 hour. The enzyme was stopped by 200 mM Pefablock, and centrifuged again. The supernatant was removed and the pellet was recovered and subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). SDS-PAGE was confirmed by western blotting using a polyacrylamide gel, in which the separated proteins were transferred to a polyvinylidene fluoride (PVDF) membrane in such a manner as to be able to be separated in proportion to the protein molecular weight regardless of the amount of protein charge. The protein transferred to the membrane was made to be 5% skim milk in PBST (PBS containing 0.001% Tween-20) and blocked for 1 hour. The membranes were washed three times with PBST for 5 minutes each, followed by primary antibody reaction with anti-SAF32-HRP (1: 400) for 2 hours, and then washed three times with PBST for 10 minutes each. After secondary antibody reaction with anti-mouse-HRP (1: 2000), the cells were washed with PBST three times for 15 minutes each, and then images were obtained by X-ray film using ECL (Enhanced Chemiluminescence) solution.

As a result, as shown in Fig. 1, the transformed prion-infected mouse was confirmed to be infectious, and protein hydrolytic enzyme (PK) treatment resulted in truncation of the amino acid end, resulting in reduced protein size, unglycosylated, It was confirmed that bands were detected in the form of mono-glycosylated and di-glycosylated sugars (Fig. 1).

< Example  3> modified prion ( PrP Sc ) Isolation of Mouse Brain Tissue Protein from Infected Mouse

Proteins were isolated from the brain tissue of transformed prion-infected mice prepared in Example 1 and classified and analyzed by function using in-gel digestion method.

Specifically, the 10% brain emulsion solution obtained in Example 1 was dissolved in RIPA, centrifuged, and the pellet was taken out by one-dimensional electrophoresis. One lane was divided into ten pieces, -gel digestion. Decolorized with 50% acetonitrile, reduced with 10 mM dithiothreitol and alkylated with 55 mM iodoacetamide. The peptide was cut into a peptide form by treatment with trypsin for at least 12 hours at 37 DEG C, and the peptide was extracted with formic acid. The sample was separated by liquid chromatography and analyzed by tandem mass spectrometry. Peptides were identified using SEQUEST based on the mouse protein database and the biological processes were analyzed by classifying identified proteins using functions of Cytoscape and Gene Ontology (GO) .

As a result, as shown in Fig. 2A and Fig. 2B, the expression of the protein in the normal group (Fig. 2A) and the expression of the protein in the infectious group (Fig. 2B) ).

< Example  4> Prion  Screening of disease candidates diagnostic indicator substances

A node having a p-value < 0.05 in the biological pathway analyzed in Example 3 was selected to preferentially select 317 proteins. Among the proteins expressed in the normal group, seven proteins having an average score of 20.0 or more, a protein having a spectral count of 3.0 or more, and a protein having a spectral number ratio of 2.0 or more were selected. In addition, we selected 36 proteins specifically expressed in the infected group with the same criteria as those of the normal group among the 317 proteins. Finally, considering the ratio of the number of spectra in the normal group and the infected group and the protein not reported in the prion disease ACSBG1 (acyl-CoA synthetase bubblegum family member 1, gi_number: 16716465, protein molecular weight: 80374.56) protein was selected as a candidate diagnostic indicator substance. ACSBG1 was not identified in the normal group, but was scored as 70.2 in the ME7 scrapie infected group and 7 as the number of spectra.

< Example  5> Identification of Candidate Diagnostic Indicators by Biological Verification

Expression of the protein selected in Example 4 was confirmed by Western blotting.

Specifically, in the same manner as in <Example 2>, normal and infected mouse brain emulsion was used for sampling and blocking. The antibody was diluted 1: 200 with ACSBG1 (sc-130090; rabbit polyclonal; Santa Cruz Biotechnology, USA), washed with PBST three times for 10 minutes after primary antibody reaction at room temperature for 2 hours. After secondary antibody reaction with anti-rabbit-HRP, the cells were washed with PBST for 15 minutes three times, and images were obtained using ECL solution using ChemDoc XRS (Bio-Rad). For the comparison between the experimental groups, the first antibody reaction using β-actin (1: 1000, cell signaling) antibody was also performed simultaneously.

As a result, as shown in Fig. 3, expression at 80 kDa, the expected molecular size of ACSBG1, was increased in ME7 scrapie-infected mice compared to the normal group (Fig. 3).

<110> Korea centers for disease control and prevention <120> Expression validation of protein expressed uniquely by scrapie          prion protein and use for development of biomarker for the          diagnosis of Creutzfeldt-Jakob disease <130> 2015P-01-044 <160> 2 <170> Kopatentin 2.0 <210> 1 <211> 721 <212> PRT <213> Mus musculus <400> 1 Met Pro Arg Gly Ser Glu Ala Gly Tyr Cys Cys Leu Ser Arg Asp Ser   1 5 10 15 Asn Met Pro Asp Ser Arg Asp Asp Gln Gln Gln Gly Ala Ser Leu Gly              20 25 30 Thr Ser Gln Asp Asn Ser Gln Thr Ser Ser Leu Ile Asp Gly Gln Thr          35 40 45 Leu Ser Lys Glu Ser Pro Ser Gly Leu Glu Leu Ser Ala Pro Glu      50 55 60 Lys Ala Arg Ala Ala Leu Asp Gly Ala Glu Ala Leu Trp Thr  65 70 75 80 Thr Arg Ala Asp Gly Arg Val Val Arg Leu Arg Leu Glu Pro Phe Cys Thr                  85 90 95 Gln Arg Pro Tyr Thr Val Gln Met Phe Tyr Glu Ala Leu Asp Lys             100 105 110 Tyr Gly Asn Leu Ser Ala Leu Gly Phe Lys Arg Lys Asp Lys Trp Glu         115 120 125 Arg Ile Ser Tyr Tyr Gln Tyr Tyr Leu Ile Ala Arg Lys Val Ala Lys     130 135 140 Gly Phe Leu Lys Leu Gly Leu Glu Arg Ala His Ser Val Ala Ile Leu 145 150 155 160 Gly Phe Asn Ser Pro Glu Trp Phe Phe Ser Ala Val Gly Thr Val Phe                 165 170 175 Ala Gly Gly Ile Val Thr Gly Ile Tyr Thr Thr Ser Ser Pro Glu Ala             180 185 190 Cys Gln Tyr Ile Ser His Asp Cys Arg Ala Asn Val Ile Val Val Asp         195 200 205 Thr Gln Lys Gln Leu Glu Lys Ile Leu Lys Ile Trp Lys Asp Leu Pro     210 215 220 His Leu Lys Ala Val Val Ile Tyr Gln Glu Pro Pro Pro Lys Lys Met 225 230 235 240 Ala Asn Val Tyr Thr Met Glu Glu Leu Ile Glu Leu Gly Gln Glu Val                 245 250 255 Pro Glu Glu Ala Leu Asp Ala Ile Asp Thr Gln Gln Pro Asn Gln             260 265 270 Cys Cys Val Leu Val Tyr Thr Ser Gly Thr Thr Gly Asn Pro Lys Gly         275 280 285 Val Met Leu Ser Gln Asp Asn Ile Thr Trp Thr Ala Arg Tyr Gly Ser     290 295 300 Gln Ala Gly Asp Ile Gln Pro Ala Glu Val Gln Gln Glu Val Val Val 305 310 315 320 Ser Tyr Leu Pro Leu Ser His Ile Ala Ala Gln Ile Tyr Asp Leu Trp                 325 330 335 Thr Gly Ile Gln Trp Gly Ala Gln Val Cys Phe Ala Asp Pro Asp Ala             340 345 350 Leu Lys Gly Thr Leu Val Asn Thr Leu Arg Glu Val Glu Pro Thr Ser         355 360 365 His Met Gly Val Pro Arg Val Trp Glu Lys Ile Met Glu Arg Ile Gln     370 375 380 Glu Val Ala Gln Ser Gly Phe Ile Arg Arg Lys Met Leu Leu Trp 385 390 395 400 Ala Met Ser Val Thr Leu Glu Gln Asn Leu Thr Cys Pro Ser Asn Asp                 405 410 415 Leu Lys Pro Phe Thr Ser Arg Leu Ala Asp Tyr Leu Val Leu Ala Arg             420 425 430 Val Arg Gln Ala Leu Gly Phe Ala Lys Cys Gln Lys Asn Phe Tyr Gly         435 440 445 Ala Ala Pro Met Thr Ala Glu Thr Gln Arg Phe Phe Leu Gly Leu Asn     450 455 460 Ile Arg Leu Tyr Ala Gly Tyr Gly Leu Ser Glu Ser Thr Gly Pro His 465 470 475 480 Phe Met Ser Ser Pro Tyr Asn Tyr Arg Leu Tyr Ser Ser Gly Arg Val                 485 490 495 Val Pro Gly Cys Arg Val Lys Leu Val Asn Gln Asp Ala Asp Gly Ile             500 505 510 Gly Glu Ile Cys Leu Trp Gly Arg Thr Ile Phe Met Gly Tyr Leu Asn         515 520 525 Met Glu Asp Lys Thr Cys Glu Ala Ile Asp Ser Glu Gly Trp Leu His     530 535 540 Thr Gly Asp Met Gly Arg Leu Asp Ala Asp Gly Phe Leu Tyr Ile Thr 545 550 555 560 Gly Arg Leu Lys Glu Leu Ile Ile Thr Ala Gly Gly Glu Asn Val Pro                 565 570 575 Pro Val Pro Ile Glu Glu Ala Val Lys Met Glu Leu Pro Ile Ile Ser             580 585 590 Ser Ala Met Leu Ile Gly Asp Gln Arg Lys Phe Leu Ser Met Leu Leu         595 600 605 Thr Leu Lys Cys Thr Leu Asp Pro Glu Thr Ser Glu Pro Thr Asp Ser     610 615 620 Leu Thr Glu Gln Ala Val Glu Phe Cys Gln Arg Val Gly Ser Lys Ala 625 630 635 640 Ser Thr Val Ser Glu Ile Val Gly Gln Arg Asp Glu Ala Val Tyr Gln                 645 650 655 Ala Ile His Glu Gly Ile Gln Arg Val Asn Ala Asn Ala Ala Ala Arg             660 665 670 Pro Tyr His Ile Gln Lys Trp Ala Ile Leu Gln Arg Asp Phe Ser Ile         675 680 685 Ser Gly Gly Glu Leu Gly Pro Thr Met Lys Leu Lys Arg Leu Thr Val     690 695 700 Leu Glu Lys Tyr Lys Asp Ile Ile Asp Ser Phe Tyr Gln Glu Gln Lys 705 710 715 720 Gln     <210> 2 <211> 57006 <212> DNA <213> Mus musculus <400> 2 tgcagcggaa aaaccaggaa gaaaaggaaa gttttagtcc tgagcaattt actgcctagg 60 aacattctga gcttctgaaa acagagagga aatggggccc tccgggtcac ttcaggcacc 120 gcccacctca gccccatgcc cgccccccgc tgccccctca aggccagcct ctcactgtgg 180 gagaccgcac tgtcttcggt gaatgcacac agcagatgcc acgcggttct gaagcaggat 240 actgctgtct gtccagggac tccaacatgc ctgacagcag agatgatcag cagcaggggg 300 ccagcttggg gacctcccaa gacaactcgc aaaccaggtg ggttcagtgc tccaaggtca 360 gcacgtgggg cttggggctc catatcacga tgtgatgctg gaggacctga agcaagctgc 420 ccctctttct gagtctcggt ttcttcgcca gatctctgtg aaggaaaaag agcagggcag 480 ggacatttat ccttcggagt gtgattcagt gtgtctgttc ctctgttggc cgttagagat 540 tcaactgagg tacaagtcac attggtatta agaaagactt ttttttcagt cagcaagcac 600 gttcctttgt ctccccatgg ttggaggtga cctggtatat ggtaggagcg ggcaggcaat 660 gccgaaaag ttcgagggag cacccatggt ttccattcca gcctgctatg accttggaaa 720 gtctctggcc ctctctgtcc gtacctgaac agagattgag gttatctctg atatggagga 780 actgcatttg tgtcccataa gagagcaaga acagaagagt ggaagatagg gctcagagag 840 agcagtgtgc taaacagctg ggctgtctac agggtaggtc aaggacaagg aatctgcaag 900 gtttgaaaac gctcaagaac atgaccactg tccttgtaag gactatgtgg gaagaggtgt 960 gacagatgag aagacatgtt aatgtgcccc attaggtccc ctggacaaac tcatcaggag 1020 agtgggctgc tgtttaccta gaaacagtat cccttccccc tgtatggatg gcaagtgtag 1080 gggagcaact gtggccctga acaattctat gctcttggca taggaaggcc tgggccctgg 1140 aggttctggg tctattctgg gtagcatcac tcatgatgtc atgaggtcag tgagcagggg 1200 ccatcccgac atcccaggtg atatttccag atgtctggaa agttcagctc tcaaaaatga 1260 agaatagcgt ctgctggttt tcaaagtccc ttgttccact tagtgagggc tggtgagttt 1320 gtccagagaa cctaataggg cacgcatgtt aagtcttccc atttgtccca tctctttctg 1380 catagggtca gtagctccag tgtttgctga agtccatgta tcgggtctat gttgacaggg 1440 acttagccaa tagggaggaa ttttgaacat ctgagctcac aggctctagg tctgcctcac 1500 tgataacaca gatagaggtg gtcaaacctg ggagccatct tcagcttggg cagcaaagga 1560 ctctgtcgtg ggaaggagca caggttcagg aggtccctgc aggacagaca gggactggaa 1620 gccagggtta gagtgtcagc tctatcgcct gctttctgag caacattgag taaattgttg 1680 acgctctccg attttcaatg tgtctctctg tgcaattgaa gatgaccttg acttcttcct 1740 tgaccttgat catatatgaa gatggtttca gagatccagg ggcacacaca ccacagacac 1800 agtaacccca gaacaaatcc cagggcatct tggcagcaga ctgcaaagca gacttgagct 1860 ctagtcgggc tccgctcctg acttattctg tgactcaagg caagacatac tctctctgag 1920 cctcactttc cccatctatc caacagggga agtgaatata ccctggcaac cttgcagtgt 1980 aactgcaaga agagaccatt gatctcttat tggagtcact gaagttgcca catctggagc 2040 caggcacagc tgcttctggg tagggttgga agtgcctgaa acctcctatt agctgagaga 2100 tcctagcttc ttcttccctc cctcagctgc agaggggttg gctggaggtg gaggaacagg 2160 cggcattggg cagctgaggg gtgcctagaa ctggtatgct gcttcttgag gctcatccat 2220 ggctgtaggc catgccattc attcaccaga aactttactg agcatgccct gggtgattag 2280 atcttggtca tgtcaagtcc caggcttggg agaaaaattc caatggaaga aagtcctcag 2340 aggaacccaa gaagaagaca attttgaaag gttatggatc ccagggaagt aaccagtaca 2400 ggggaatgca ggagtggcca tcagaccagc aggctgaggc caggctcatt cagagcctgg 2460 tctagagaat ctgaggtgcc ctacttgtga tagacagggc caggacagcc agccagagag 2520 gtagaggaag ccaatagggg catctcttca tccttggaag gttcccaagc cactcatagt 2580 aggaagtaag gaacagatat ggtggtggcc acaaggggac tggacatgag gtagctggga 2640 gaggtgaggc agaagattgg tctgtagaat tgctgcctcc aaactgcccc caacctcaca 2700 cacgagaaat acctgctata ctcttagctc tgcctgaaaa aaaaagccag tgttgggagg 2760 gccccccccc cagaggccat catggaagat agtccaggag gctgcagcaa gctaggcaag 2820 aaaggctctt ctctcagaca aagctcgcca aggataacag ggagatatca ttccatctca 2880 gggacgtgct aaccttagag gaaccatcag catggaatgt tatcacacgg tacaagtgtc 2940 ctcggcactg ggaacactca gggccttcca gttcctcttc acgccactgt ctatttgcct 3000 agtgcagaag tgtggcctgc ctttcttttc ttcgtgcttt gccatctcta ttgttttcct 3060 gttttataac atactcatat actcaaaatc atttgttgtt gattacttag gaattgcact 3120 aaacacgaga gtatatcaga gactacaaag caagcctctg ccttaaggag tttacatgca 3180 tgccaagaag aaagacatta ataaacaatt atgcaattag tcattaaact tacagtttaa 3240 tgataccaaa aagtaatttt aaaatgtgaa gaaaaaaatg ctgatatgaa tgtgtgtagt 3300 gtgtggtgtg agtgtaagtg tgagtgtgtg tgtgtgtgtg tgtgtgtggt aggtattagt 3360 ttggctttca tgagtacatg ggttctgtac ccagtaccaa agggcctggg agactgatga 3420 gagtgttcat ccagaaaagt tgcttgccac caagcttaac aatttaagtt caatcccaga 3480 acataaatgg tagaagtaga gaaccagcca ctgcaggttg tcctctgacc tctacatgca 3540 aataaatttt ttgatgtagt aaagaaaata aaggaaaaaa tactcaggac cagggcatta 3600 tacaccttaa atggtaaaga tgtcaactgc tatgctgtgt ttattttacc acaataaaaa 3660 atggttttta atgttggcaa tcaaaaaaat agaagtaatg taatagagat ccaatttagt 3720 cttggagatt aaagaagact tgaaaatctg agacctgaaa aataaacaga atctcactag 3780 gcaaagtgtg ctatgaaaga ggggaaggac cttggtgttc gaggtcagtg gaatgaggca 3840 gagctggaac tctaacagcc ataggaagtc aaggcattgg tatcaagaga ttccaaagga 3900 cagtatctgt cagggaagtc agagaccctg aggcaaacta ggcctggtcc cagacacgtg 3960 aaagtaattt gaatgtcttc agactaatcc aggccaactc tgggtccttg taggttctct 4020 gcttgtgggt gcttgtgggt gctgggatca aagctaatca ctgtgccctt aggcaagccc 4080 ttctgaacaa cagataccct cccaggtctg atgggttctt acccattctg ttataacacc 4140 cilgagaat caatctattc taacccaaat acctccaggg aaggcgaatt tgctaactaa 4200 tgtgttatct ctgtaaggac agtggtggta gaagaaagtt ctgtttcaca ctgagtcctt 4260 taagacaccg ctgtggtgca gaaaaaatgg aagggtcaga caatgagaag ggggtggggc 4320 aggattgtgc gatgctcttt taggcaccag gtgactgtca cacttaggaa ctcaaagcag 4380 ctgggattac ctgcataaaa caggacccat caacattcag tcataggtga gggaggggct 4440 catggggctc gatccctcct ccaggtgtga ttgacagctg tgggatcaga agtcattgtc 4500 tttagtggta cagccactgc atgctgcaat ggatgctaca gtgggtctct ctctctctct 4560 ctctctctca atcatgcaat catgataatg tggagctgga gagatggctc agtgtaagag 4620 cacatactgc tcttgcagag gtcccaagtt cagttcccct cacccagtat aactccacct 4680 tcagggggat cttacacttc tggccaccac aacacacaca taattaataa taataaaaat 4740 cttttcaaat gatagtagaa gggcacctgt tgacaagaga gacttgtggc aaggaatggg 4800 gataagctag ggaaggtggt gggtgtgacc agaatgtata agagtgtgtg tgtgtataca 4860 caatgtaaat acatataaga ttgtgattgc ccggggatcc attaatttaa aaattaatta 4920 ataatttaat taattaatta aaagaaaaat agtctcctgg attgccgact atggctcctg 4980 acagaagtat gtgtagacaa cacttaggag cagggtgctg ggaaactata aaggttatca 5040 caggagagcc atctggactt gctccctaaa ctacttctca cagaatgtcc atctgaagtg 5100 ctgagtgggt gggtggggtt ttattaatgg aggtaaatgt ttcatacggt gaaacaaact 5160 gacttcaagg tgcacagcgt gtgattggca acccaagtgg aatatagaac atacccatca 5220 ttccagaaaa gtccttcatg cctatccact tctctgcctg tttttgaaca tcatgtgtat 5280 aagccaacct gctcagaatg tacccttctg tgcctggctt ttgttgcacc ttacgcttga 5340 gatgtcgttt gtgtcatcaa gtgtttcaac agtacattct ttattctttg cgtgtgccat 5400 gagaatttct tctttattct gtcacggctc atttgagttt cctggtttat ttggtgtttg 5460 tcacatggcc actgccacca ccctgaactg ctgccacctt cttcagaacg agtgtgattg 5520 tactcggggt ctgcttggta atacaaggtc atcttcctgt cctggggtcc ttaatcacac 5580 ccagaaaact ccatcctccc aaaaatgtca tatttaaaag ttgcttgtga ttaggatgtg 5640 ggtaatccga gagacatttt gattactatc ctggtcttgg ctggaggcct cagttctttg 5700 ccaatggttt actttgtgag ttactcaagt gtcctcactg cacggtggct ggtcaccagg 5760 actactagag aatcatccac atgtctggag gccatgctgg gaagtgaccc tctggaactc 5820 ctcaggatac atgtttttct atgggccttc tctgcatgat gtcaccatca tgccttccag 5880 ctaggtagac ctctgacaca gtgggtcagg gatcccaagt cacatgtgtg gagagagtca 5940 gatagaaact atttcttttt cagacccagc cttggaagct aggcatttcc ctgatgtgct 6000 tgttggaaac taggcattgg tcttttcctt attaaggggt gctggtggga ttaattagac 6060 ttcacctact gatggaatca gtgttcagaa tttacaagat ggccaggcat ggtggcacat 6120 gcttgtaatg ccagccctca ggaggcatga gggttggaag ctcaagctag gatgggctag 6180 tgagggagat cctatctcaa aagagaagtc tccaacatgc tttcaatcta tggtagcagt 6240 ttttgtgaca agtcctccta tctttgattt gtaggaaaac agccattctt ttactatcct 6300 tttaaaatcc aaatgttcgc tctagccaca ctgacatccc tggtctctat ggtttcctgt 6360 ggccagtggc cagtcacttc tgcccttgtt ccttttgtgt aatgtgtcct ttctctctgt 6420 cctccatttc ctttaaaata ctgtcctcgt ggctttttgt ttgtttgtgt ttgtttttgt 6480 ttttgtttca aggggtttga ccgtgacggg ttcgagtggg atttttcttt cccttttgtt 6540 aaagcccagc tgaggctcag tgacatgaaa attctagatt ttcctgtgta atttgtaggt 6600 taatttttta aaccaaactt aaaaaatata gacatggttt cttcaaatat gtctctatgc 6660 cccattctcc ctttcttctc aggaatttgt gtgtgtgtgt gtataatgta tattggagca 6720 ttaaatatgt atgtattata aatgtatgtt aaagtattta ataggtcact ataaatccct 6780 ttattttgaa aaatattatg acttcactga aatacagtca catatgtgaa atttactccg 6840 ttaaagtgta tagtttgttt ggtggtatat tcacaaggat gtgtaaccat aattatcacg 6900 gattttggaa tactgttttc acccggaccc ataagctctt tgatgccttt aatctgagga 6960 agccactaat gtgtctctgc cgagacgttc tactgagacg agctgatacc tgacctattt 7020 ccattttccc tctcacttag tgttaatgct ttcacaattt actggtgata taacatttat 7080 tatcatctcc ttccctttgt gtgggtggga cggtcttgag acaggctctc tctatgccac 7140 ccaggctgac cttgaactca aagcagttct cttgccacag cctcccaagt gccaggacta 7200 aaggtgcagc agtttttcat tccttctatg gctaaataat attccttttt atgttacagc 7260 ttgtttaccc ccttccatcc accgatggac acttgggtgt tataatttat cagactacta 7320 tgaatattca tgtctcagtt tgggtgtcaa tatctatttt tggttctcct ggatatactt 7380 ttatgaggga gttgctaagt cagatgaaat tcaatacata acctttaaat ttcttatttc 7440 cccttttgag acagggtgtc actatggact cttagctagc ctagacattt ctgtatagac 7500 caggctgtcc tcaaaatcag agacctgcct gcctcccaag ttttggggtt aaagaattca 7560 tcccatactc agctcaacac atgctcttga gaagcaggaa gagagaagct ggcgtctgag 7620 cagttgcagc cgcaagggct ccaggacttt cacatctttg ccaacacacc tccccttttg 7680 ttacagcaag cctggcaggt gtaagctgtt atttcgtgtg gctttgaatt gtgtgtccca 7740 atggctgatg ttaagtaacc tttaaagatt ttgtcaatgt ttatacttct tcagagagaa 7800 gtcttttcaa ttctcctttg tttattgttg attctgttta ttattgagtg gcaaaggtta 7860 gtttttgttg gtattggttt ggtttggttt ggtttggctt ctgtctgtag ccctggctgt 7920 cctggaattc actatgtaga ccaggctggc tttgaactca cactgatctg cctgtctctg 7980 cctcctaagt gttgggatta aaggcatgtg ccaccatcag ccttggcaag ggttctttac 8040 atattctaaa gacaagtctc ttgaccacat atctgatttg gagaatattt tctcttattt 8100 tcctgggtta actttttcgg gttcttgata gtgtcctttg aagctcagat agttttaatt 8160 ttgattacgt ctaacttatc tgcttttctt ttgttgctta tatttttggt ggcaagtata 8220 agaagtcatt gcttaattca cggctataaa atttacttct ttcaaagttt ttatagcttt 8280 agctttttca ttaaataatt gagttcacac tgaataatta gatcacataa tttatattag 8340 tttggcccat cttgaatgtg tgtgtgtgtg tgtgtgcgcg cgtgcgtgcg tgcgtgcatg 8400 tgtgtgtgtg tgtgtgtgtg tgtgtggttg gttggctgga gaagagcttt ggcttctttc 8460 tttctcagat gcctcagcct tactggaaag attcttattt ctctgttagg ttgtcttctc 8520 atcttagtga aaaccagttc attgtaaact gacaataatc ttaaattaat ttctgagttt 8580 tgctcaactg cacttgcccc tgtgaatact ctgtgatacc ttgtgtgcca tagttttgat 8640 tacagagggt attcaaaatg ttctggaatc aggagatata aatccttcaa ctttgttcta 8700 ttctgggtct tgaattttct tttttaaaaa catcatcatt attactatca ttattactaa 8760 ttattttatg tgtgtggata tcttgactgc gagtatgtat gcaagtgtat gtctggtgcc 8820 catggaggtc agaagacagc attggatttc ttggaactgg agttaccatt gtgagatccc 8880 atgtgagtgc tgggagttga actctggtcc tttggaagag ctgccagttc tcttaattgc 8940 tgaatcatct ctcttgttcc tggctcgttt cagtttttgc aaggatacat cccaggattt 9000 ttatagagat tatattgact tttggccaat attaaatcaa tcaatatctc tgtctctctg 9060 tcttggtctg tctctctctt tgtctctgtc tactcttatt aattttggag ccagggtctc 9120 acaatgtatc ccaggttggc tctccatcct gccaaagccc aagtgctgga ctataggtct 9180 atgccataat gaaggctgca ctaatgcttt ctcactccta aggcatctat ttagttcttc 9240 agcttctctc agtaatgttt tgtcctactg aatttaaaat ttctttcctc tctatttctt 9300 ggatatttct atactttcta ttatttgtct tcaagttcac taacacttgt tttcaatttg 9360 gatctgagac catctggtga acccttcatt tcagagattg tacttgtctt acattttgtt 9420 gtttggtgga gacttcctcc ctgggggatg gggagaaggc tcagtcagtg agaagctttg 9480 ctgtgctcag agagaaccta ccatgagctc tcccagtgcc cataaaagtc tgggtgggca 9540 tggcagtcct cttgacatcc tagcattcag gaggtagaaa gagggaatcc tgggacaagc 9600 tacctagcta gactcaccca attgataagc catgagttca agtgagagac cctggctcag 9660 tgactaagat agagtgtgat gggagaagac acccagtgtc aatctctgac ctccacaggc 9720 accacacacag gcatccacca aaacacactc aagtataccc agacacatac acacacacac 9780 acacacacac acacacacac acacacacac acacacacac acacacacac acacacacac 9840 acacacacac acacacaca acacacacac acacacacac acacacacag aggggggagg 9900 gaggatggta cctatccccc cctttaattc tctagtttta ataagtacta taaagtcctg 9960 gtggatatatt cactaccatg tctggaagag tggaaaaaaa ctttaaattc agtaggacaa 10020 aacattactg agagaagttg aagaactaaa tagatgcctt aggagtgaga aagcgttggt 10080 gcagccttca ttatggccag agcttgggct ctggcagaat ggagaggcca acctaggata 10140 cattgtgaga ccctggctct aaaacaaaca gcctttgttt ctatttctgc tactgccttt 10200 cccccttgat tgtgtatatt ttctgatata tagttattct tgtgtggatg gctgaaagga 10260 ctatgtgact gttagtttga aggatttaca aacacactag actagtgtct tagccaccat 10320 gtatgctttt cccagtcagt tgtttctagg aaattccaca ttgctgagac atacagaggg 10380 agtggagttg gggtcttcac ggagctgggt gcagggtcag ggatgaccca gcatgcacaa 10440 gtgaatagct caactttatt caagtcccaa caccagaaaa caaaataaaa ataaaaccca 10500 aaaaacccac catgaactct ggagtctgct cacctgggtg ccaagtgtcc ctgtcactag 10560 ctatgtggcc ttccacaatt tatcacaata cacttccatt acctcataaa atgggggtta 10620 cggtcagtta agacagctaa atgaaccagt cctataatgt aacaggaaca ttatctggta 10680 tataccagta ccttggaaat gctagagttg gagtcccctc tgcggcccaa aggcactcta 10740 accacccctt gcccgccaag gtctgagaag tacaggctgt ggttgtgggt cacttctttt 10800 ctcttctttg actggtgtgt accccagggc aggggtgaac cctgagaccc agcatcagca 10860 cactctggt acactgacgg aaggcttccc tcggcagaga ggtttatcta gacctggagg 10920 aacaggcagc acattcctcc cctcgtcccc ctgggagaca agcagagaat catctttcat 10980 cagagctggg atgctagccc acccatgtac ttctctctag atgaggcgac cgaggccccg 11040 agaagaaaag gagcacagtc agagtcacgc attcagcact tcaaataaaa atggccatca 11100 gtggagagtg ggtgatgctc atgtcatctt tgggctagaa ttatgtcaat gttgagcctc 11160 ccctgaggtc atcaggccca caaattgtat ctatcatgga tgccacatgc agctaagcct 11220 gaaaccctac gtgtgatgtc ttgagcctat ctgatagagc caacaactcc tgcaagttgt 11280 tctttgattt ccacaggtgt ggacacatgc ccatatatgg gcatgcagag aggggggagg 11340 ggaaggaaga aaaagaagaa agaaaggaag gaaggaagaa agaaagaaaa gaaagaaagg 11400 aagggagaaa gaagaaaaaa gacaagatgg ctcagtgggt aagagcactg tctgctcttc 11460 caaagatccc gagtccaaat cccagcaacc atctgtacag ctacagtact cacatacata 11520 agaagataga taaagctggg cgtggtggcg cacgccttta atcccagcac tcgggaggca 11580 gaggcaggca gatttctgag ttcgaggcca gcctggtcta caaagtgagc tccaggacag 11640 ccagggctat acagggaaac cctgtctcaa aaaaccaccc ccccccccaa aaaaaaaaac 11700 ccgaacaaac aaaaaaagaa gatagataaa tcttttttta aaaaaagagg cagcttcatt 11760 aggattttag aactggagtt atggcctttc ttattattca tagcatctgt cacaacactg 11820 gatatctgat atgaagtcag gaatcattgc agatcatgag agtgcccatt ctcgactggc 11880 ccttttgtcc ctgccctgtc ccactccaat caccagagtg gcccaacatg tccagggtat 11940 ctgagctcta gagcccacct tcctgagtct gtattgagtg tgcttcttca tctacaaaac 12000 tcggacagca gtaaaatttc atcgggccgt tgtgaagatg gatatatcag atgtaaacat 12060 ggaaagattt gcacaatacc caatgagtgc ttggtacagg aaagtgaccg gcatatactg 12120 atcttctccc ctcatacttc ccctgccctc tcaggcttgc catcctggca acaggcaaac 12180 ttgcagcttt aatctcttag tttagcccag gccctaaatt gtcagtcata gccaggagaa 12240 ctgggaggcc ctttatctgt gacatctcag aaaatggagt accagaggag aatcagtcac 12300 atggttctga ggtcacatgg tcaaggcaaa gtctgggcca gaatccaaag attaatgttc 12360 agagaactcc gccctgggta tgattaattt gctatctgac tcctttggga gtttgtagta 12420 gaacttgatg attggtatat ttatttccta acacagacaa tgggatttag cttaacaatg 12480 ttgaacactt ggtctacata ctgccatctg ggttataagt gaactggcta gatggctgcc 12540 ccacccttgg agatgggagc caagtctcag ggaggatgaa tgttagaggg tgaggcctct 12600 cagtaaacaa cataatcctg ctaggatcca gtcaatcaag ggccccagga tccccaatac 12660 gttccttctg gtggctctgg gcatatgtta gatcatgtat gccttaccgt aaccctttcc 12720 cagagacaaa tactatcttc actttggccc agggccctat ccatgaggtg ataactggat 12780 ggtccacagc ctggtctgtt acccatgcta gcttcacctg ccctacccct tgtgcctcac 12840 atgaagtaat aataatcata gcttcaaccc tactcaaaat tcttcctgtg ccctctccag 12900 ggctgtctcc acagtatctc cctccccata gccttggaca catacatgct cagcagccat 12960 taattatctc tacaccacat ccagtttgct cacctctgtc caaataagtc catatgccat 13020 ctggatcata gaaggcatta actcaatact cagggtggga aggaggaccc cctagaagct 13080 gtcaattact aggtcaggga gaggggttat ttctaattct gagagtaagg actgagttaa 13140 aatttggtct cctggcaatt cctagtgcct aaatgctaac tgccttggac tagccaccca 13200 gttctaggaa gagctgtaac aagagaggac cctctcaggg tgcttcttgg tgcaggtagg 13260 acttggagaa gaaggaatct gttggaggag gctacatact ccccaagctc cccgaacctc 13320 atccatctgg aaaccctatc cagccatcca atgggctgaa ggctgaggct acctttgcct 13380 tcaagtacag acattgtcac tagtaaccta cagcgagaca cagtgttgat ctcaaccatt 13440 atgccatttc taagccagca ctgggacatc agaggtggtg ggagatgggg acatgacacg 13500 ggacacagac tccattggta gctctccagg tgggaggaaa gtctgaaggc ttttagtgcg 13560 atgaggtcgt caacagggcc tggacaccta aggtcccaca gattccagaa atagcaagag 13620 agttgcatga ttttgaaaat ggggtgctgg tagttttctg ccacccaagt tggcagagga 13680 ccttcccaca atctcctacc tccattactg tccccttccc cttcttttgt gggtaaaatt 13740 agggtttaaa gtctccctgt gctggctagg gcacctacag gctctcatct cagcccagga 13800 gtatttgggg tttggagttt agctcccagg ggataactga aactgaagat atggttgaat 13860 ttctcaaggg ggacaaggaa cagaaccatt taaggagcaa ctgaaagggg aggtgggcaa 13920 gggctctcct ggcagcagga ccctggtttt aaaaggcact aaaggagtca ctgcagctgc 13980 aagtgttagg ggcagtgagg ggcattagga agctgagggt gagatccctg gatgagggct 14040 ctcagaggta gtgtgggtag gatggatgaa tctgtgatgc aagtgccaga taaacctgtg 14100 tcctggctct gtcatcagag ccagcttaaa catgctagct cctcctcttg ccagctgtgt 14160 caacaacaga gcaagtctgg agctctctgg gcctccactc actcatctgg aaaatggggc 14220 tattaataac actgacatta caaggttgag gcggaggata caggaaacca gcccagccca 14280 tgtgacaagg tatgtgtact taggaaccgt gaaaatggaa cacttctcca aaagaaaaaa 14340 aaaatttgaa tgaacaagtc agtcttctgg gccagcccaa gaggccccag gacagcctct 14400 caggtatatg tcccagtagt agaagtaatt tctccaaagc tctgacaggg atccaacaag 14460 ctgtctctag tactgcacta acaggtccct gtcttgcttt tcgccccact tatgtaagag 14520 acacataggt gggcagctgg gtcctatgcc atgtaggtca gaggtatctg gagccacacc 14580 ttgccaggag ccccaaggca ggctggggcg ggtccccatt cacgcccaac ccttgtggct 14640 catcacccag aacctgggcc acagcgacag ccagttctca gattcaccat gccttagctg 14700 caggagcagg gggtacgcag ccctgagcct gctaacgagg aagagaaagg tgagcgggga 14760 gtttgggtag agcctgctag gcaagtcctt tactgtgagt cttctggggt tggggggtgg 14820 ggtctgtggg atggaagtga gggctggaga gagttctcag aagcctcatt aaggcagtcc 14880 atatagacca aggtgaagct gggatatgag gtcccatgta caaggaaggg aatagtattg 14940 caaagtgtgg cctcagcctt ctttgggggg ccccagcatt tggcaccact aggttgtgga 15000 gaaaccattc ctctctttgg gatttttaag atgcagggtg gcactgaggt aacaccttca 15060 ttaatagcat cctcaggatt tccagaagct tccattccct ttgcctctga gcagagaaag 15120 aggaacactt caagtaccag aggggtactt gaacctgacc ttgaagaaca gtggtcagaa 15180 gtctttgcaa agcaggagtc ctggttgatt tgctcttgca ggtgcaactg gctctaggga 15240 atattgagcc cagagcctca aggcagccac ctccagtgag tttgtgttgt tcccagattg 15300 aaggagcatg taggtttagg aggcagtaaa ttacaggtga ccagccccag aagcagcgag 15360 cgaggcatgc ctcaaaggag ggcaactacc cattgccctg ctaggcctgg cagtgactca 15420 tcatgtgtct ctgttgcagg gcggtctggt cgagttgggc ctctctgggg ctgcggtggc 15480 tttccccagg ggacgctgtg acacgccctc tgagcttctc cctctctccc tgtggagctc 15540 gaggggtaca gccaccccac agggaccatg ccagaagtgc ttcttctcag gtctgccagc 15600 tccatcctga ggaccgtgta aggaaccagg gtcccaccat cctcctgcct ccatccccat 15660 cgccatctcc tgtgggggag tacttagcac agcacctgag tcccagacct ggatttaagt 15720 cttttcctcc tactcactca ctctctccac cttactgtcc aggcttctca cttctcaaga 15780 gatccatgac aatcctccaa gaatatgatg acagatgact ctgtactggg cacttttcct 15840 gcctcatcct ctcacggtag cacagggcat ggattataaa acagctcaga gacacagggt 15900 gatttgacca tagcatacac ttgaaaagtt agttgaaccc tagcagatgc acagacgcca 15960 tgctaggctg tttcaagaaa acatttattg agcacccgtt gcatcactgc tgagggctaa 16020 ctgggaagaa taagaatatg taagaaatgg atcttcactt gcgagggatt catacagaaa 16080 gcaagggttc tcctttacct atgtatcaca tacagtgagg atgtggtgat ccaaccaaca 16140 tttactgggc taccacagtg atgactcatg gccagagaac agacgtgagt tgggaatgac 16200 cctactttcc aggagcttcc tgtgtttgag cttcaacaac caatctagaa ggtgttgggg 16260 gagagagagg agcgcttcct ggaggaagag ttaagaaggt tagagaggtt ctgcgtccag 16320 atggctccca ctagtctcct gcgaggcaag cctgggtgcc tacctgaggt ggaatgacgg 16380 ttaagaggaa ggcggtaagc aggccctgcc acacccctct gctctatatg caccatagca 16440 agttgctcaa acatacctgg caaggcggca aagaacccca agtcttgatt tttttttttc 16500 tggtgaaacg cacaacactg tgcttgtatg agaagaatgc tgggtaggtg aaagcagcca 16560 gtttcctgcc atcttggaat tcttcctcct tggaattcaa aacacagaca gaagtgagtc 16620 agctcaggtg cttctcccac tggccactgt cattttatta cttgaggaga cttcctcctc 16680 ctggaaacca gcactgattc cagaggaatc taaccttcca gagtgtcatg tgtaaatggc 16740 cggctcctat cctcattact gtctgtagtg tcatctcacc tgctgggctc accgcaggtc 16800 acctgtcctg gaaagagcag acagtcaaat ggaaagcctc catttttttc cacttattac 16860 tggggattga acatgagacc tccctcacac atgctaggtg cgtgctctgg catcccaggt 16920 ctctaaaatg tttttttttg gaggataggg ttttgataaa actatgagcc attccatttg 16980 gggatctctc ttctggttta acctatagtt cgagcacact tggctctctt gttttctatt 17040 ctgttataaa aatgatacct tgttagacat cagcatacat agacaaacaa caaaatccta 17100 gcttcctcca gcaatgacaa agaagccaag ctgtgtgagg ctggggattc atggagaagg 17160 cacttgctat aatgcagcta aatccacttc tgccactagc aagaaacctg agtgagcagg 17220 aaggacaata aatatattct aactcaggtg attgtccctt gaagggcttt ggcaagatct 17280 ttgcaaaggg agatcagcaa tttagctgga aatttgtctg gaaaattccc tttttgactt 17340 tttctcttaa cccaggtacc ccaaattctt taagcccagg acatgagggt tgaatatgac 17400 cctcatgggt ccattcaacc atatgggcca gctgcaaggg taagtgaggt gtccccttcc 17460 accaagtcct ggaggaagaa cggggtccac ctaaagggag ggagaataga cctacggatt 17520 tgttgaagag ttcttctgca ctgggtgcca gacctcagga aaggcctgtt ctgggaagac 17580 tgggcgtccc ctcactctgt aatcaggata agtagcttag ccctcgggct tcccccaggt 17640 ggctgtcttt tcatttcctc tgtttacctt tgtgaaagcc agaaccagtt tgcctaaatt 17700 taattaggag gggaactgca cctgtacctt taatggcggc aacttaaaaa cagacctgtt 17760 gagaagaggt tgttaagatg ctgagagatt ataattctgt ggtggctctg tgctttggaa 17820 gatggatcat ctttgtgcac agggtttggc caggtcccat ctttggagga tacagctctc 17880 catgggagct catgggcttg tttgacaaca ttacagtctt tgtcctcaag tggggcagct 17940 tctacccagc agtctcagag aaccactgtg tcgtgactga ttctcccaag tcaataaacc 18000 cagaaggctc agaaatctca gctttctcgt cctttctatc ctccctggag caggtgtgcc 18060 cgagaaggtg ccaattcagt cctccacttc tgcaccaggg catcatagca acaggaggga 18120 tgctccagcc ccatggtcca ggccttactc acagtgtgca cataggaagg accaacttcg 18180 tcctctcctt ctttcccttc ccgcccctct aacttcataa cccccttttg ctttaaaaca 18240 tatctcaggt tcgtaagcag actgctgcca ggtgggcctg ggtgtgttcg aaaacttagt 18300 ttgaacctga agtaccagca aggaataagg ccgagtgtac gaagcaggta actggcctgg 18360 cttcctggga agtgggtagg gatggggcta accccaccct gacaaacaaa tgggacttct 18420 tggagccagg atcataccct gggggcatct atgaccagct ctacaggtta cccactacac 18480 acttccagta ggctctgttt acatagctat gacgatgacg tgttcctggt gttgtacagt 18540 cttgcagcgc tgtgtagttc tgagggtgtg tattgaaatc cctctgccta tctacccagg 18600 caggtgggcg tagccaggta aactctgggg agggctcaca ctgagggact gagggatggg 18660 gagactgaga tgatgacaga aatccaggag ctggtcctta ctccgtgcta cccagcctag 18720 gaggccagaa ggagctagga caggcagagc cccaggcctc cctgacccgt gtcctctcca 18780 tggccagtca gtcagcattg aaggcagacc tcatggaagg agcctcttag gattccaaga 18840 gttctagaaa tggcctctgc tctactttga gcctctaggc aagttctggg actaataggt 18900 gtgaagaatg atgtggtggt ggtgtggtag gtggggggga cttctcctac tcttaaagcc 18960 tatgttgtct tagagtggag gctcagagtc tctcaaagcc agagtccaac ccttttcttg 19020 tctggacact gactcagagg gagtcatgaa cttacacaga gacacactgc aaataagaga 19080 ccaggggaac ttgagttctt ctcctacgag ggtcaggggc ggggcactag aggaagtgag 19140 atagacacac aggatgttct aaccactgtt actcatctcc ccccaccagc tccttaattg 19200 atgggcaaac actctccaaa gagtctccaa gtcacggtct cgagctctca gctccagaaa 19260 aggcaagggc tgcctctttg gatggtgcag gtgagttttc atgaatggag tatttctgga 19320 gggagttgtg ggggctgctg ggagccacca agcagaactc tggggaagag tacatgaaat 19380 cttgctatgc ccacagctgc tgttagggcc agccatgatg gaagttcaag gttgattctt 19440 tcctcagaaa ggttggcaag gggccaacaa tgggatggcc ccagcaggca gaccctgaat 19500 gcttaggctt tggaaggaca ttaaggggcc tagccagatg actaacacct tgggagttta 19560 gccggttaa ctggggcata gggcaggcag atggatgtaa gctagcacag tttgttttgg 19620 atcaccacat gttctctcaa gtgaccttcc cctcagataa gtgtgcctca aggaaggcgg 19680 ttatcattca ccctgaaatt tatctggatg ataatgataa cacaaguca tttgttcact 19740 tttatcaaat gcacaacaat gttgttacaa aagcagcatt tcaaaataaa aaaacaaaca 19800 aaacaaaatt taaaacaaaa caaaaaaacc caacaaaaat atcccaccct gtgggcctct 19860 aagtgagagc tgattggggt tgaagacaag ttgctatcaa gtggccatgt ttaaagtggt 19920 gatatatttt tgttgtacgg ctttctgttt gagtatgcct atatttttct cgtgattaaa 19980 gtatctacaa gtttgggagt gtgggtagca atgtgtgaaa cagaacaagg cagggaagaa 20040 ggagccattt ctaaatctct gaactacaga gtcatgttag atccagggac cagagaaaga 20100 tgaggaggcc catgcctctg tctcaagttc tcgtcccttg tatttctgct gagtcagagt 20160 ggctcaaaga caacaagagg gcccgggttt ctctgctgtg tcacaggtgt cataggctag 20220 agggaagtca ctgtgctcac tttctgtagg ggaaactgag gcataacagc tctttcctgt 20280 agtctcaaca cttaggaggc tgaggcaggg tagttgctgc atatctaatg ccagccaggg 20340 acagatagca agttcccagc tagctctgtc tctaaataaa tataaaagat gatgatgatg 20400 acgacgacga cgacgatgac gatgatgacg ataaagcaat caaagaagta taatgcagag 20460 aggtcctgga acagaactcc gggcacttca atctcaggtt ccttcactga gtgggcttgc 20520 cactgtcatt gacacgctgc ctctggtgga cttgacaaga gcctggcttg agggctgaga 20580 gggtcttatg gggatgctgc ttcctttaac ccctttaacc cccctgtcag gggtaaagga 20640 ggagccagac tgtcccttgg tttcagggaa ttggtttgct aggttccaga ggaggcaaaa 20700 gggagagcct tgtgggtctc aggcagctat cactgccttg ctcccgggag ccccggcaac 20760 tcactgatcc attgtcttcc ctccagtgcc ccctccctgc atgaacccag cttgcttgtg 20820 cataagttca agtgcatcct gaggcacttg aagggcaggt gccaatacca ttggcttagg 20880 ccagggaagc cctagggctc agaggtcctg gaagtgggtg gtagcattga cattcttgca 20940 gttagagag aaggaagaga gatgatatgg aagagctcag aagggcatct ggctcccctg 21000 ggaggtacaa gtgatgctag cctggggacc tcagttgctc aattataatc acaaccatcc 21060 ttagccctgt agctgcaggc gcaggaaaaa gcaggtctga acagactgag tatgggatcg 21120 cagtgaagga cctgtctaca gcaccatctg agcatctgga gacaggcagg ataaggagag 21180 gacactggag gtagtgtagg agggtgttac aggaagggac aaagctacca tgactagtct 21240 agcaggactg tgggaatctg tcagggacag gccagccctg tctagctggc cctgtctaca 21300 ctctctacct tatgttctgg gctttaggga tgctgaacta gatgagtttc attcccgccc 21360 tctttggact cccagcctct ctacagatag attctgagag tcaacatgga tagaggaggg 21420 ctggggtttg tgggcatcct agatgtggag gacctttggc tatcctcaga cctctggctc 21480 cattagactg ttggcaggtg agtgtgtacg cacacacaca cacacacaca cacacacaca 21540 cacacacaca cacacgtaat agacctctta tcctcatggc tgtcagcagg tacccagaat 21600 aaatgggctc ctcctatagg ccccactcat gcaagccatg tccctgggga ggggaggatg 21660 ttagtggaac agcccagagc tccagatgta cctggattca gtgtttctca tcagcacgat 21720 aaaccttgtt tagacctggc atcctttgtc tccctaggtg aggaagctca tgggagttag 21780 gggggacaga ctaggctatg gtggccattg aaggaaggaa aggggaggga tcccaggttg 21840 ggtgggtggg ctgtgaagct gcctgggtgg cataacgctt tcagaagatt aaaagcaggt 21900 aggcagcagg acattacaaa ggcctgggag gggcatgtgt gggatccaga caggagagaa 21960 aatcctgtcc agcagcccat gcgactgggt ctctgttgaa gcacttttcc attttcttct 22020 gagcaacaga ggggaaggtg ggctaatcac ttccattgag aaaaggaacc tcacaggcag 22080 gtttagtgat ttttttcaaa tgttcttctc tctgtttctc tttgtctgct tctctctgtg 22140 tgtctgtctg tctttgtgtc tgtcttgcat ctcactctgt agcgcggcag tccttgaact 22200 tgcagtcttc ctgcctcagc ctcctgagtg ctgggatgaa agatgttggt cttcacaacc 22260 agaagtgatt tttctctgag ccaaacacct tactagaggc agagctgctc agggtccctt 22320 ctgcctcttt aggttctaga tagaccctgt tgcacagcag gagtcaggct agccccagcc 22380 tcctcaccac caagctgcta cagagctcct ggattcttgc ctgctggagt cagggttcag 22440 cgaagctccc tcagaggcct cttggtctcc tctagttccc aagagatttg agggtcctac 22500 attccagtcc ctaggctaga ctcccaggga gcagtggtgc caagccagag gctggaatct 22560 gacctccaca caaaggcagc cttgtgccag ggcctgagac acattttttt cctagccctt 22620 tctaggttga ccatgaaccc cagctgaaga aggaacgggg ggtgggtggg gggagactca 22680 gagagagcag ctgccagggt cacagcagat agaggtgctg cctggagtag ctttccaccc 22740 ccacccctcc ccccaggagg ttgtcagtcc tacctggagg actgggacaa gctgaggatt 22800 gggaactgga ggcgcttggc ccagcttggt ccctaggcca gggaggacct ctgaggccca 22860 agggttttaa accatttttc tagtccctta ctcttatccc aggggcacat gtcagggtcc 22920 aggctggaga accaaactgg agagacagaa gtcaacaaac aggtttctat cctatctcaa 22980 gaagctctta gccggaagct agggagtctg aaggggccca gctgtcactt taacctgttt 23040 tggggaacag gattagggga aagtgggaag ggattccccg ggttctggag aaatggtgta 23100 tctatgaaac ggagccagga gataatctat ctctgtcaac caaagacacg tctgaagggt 23160 tcatggagag acttttccag ggatcaaggg acttggcttc tcctagacat tattcagaca 23220 gtgtgcttca tgtcctgacc ctctgccctc tcctagggac tttcttttct aagcaggtag 23280 gagaattcca gacttcattc ctctgtagca ggtggtcttg gccctgggtc tcttaatgtt 23340 tttccacgct ggctaaaaac cctagagccc aagagctggg aggagctcat agctgttacc 23400 tgttcttctt tgttatcatt cagacaggga aaccaaggaa ggctctggaa tggcagatat 23460 ctgtctaggg tcactctgcc cttgacccat cactgggaca cctagcaggg gacggccagc 23520 ttcagggcct ctcctaccaa acacagagaa acagaatcat tgaactttta gcagaatagt 23580 ctatgccaca ccaccaaggt ctggacagac cactctgacc tacgtgcctg tccttattct 23640 tcatatccca aggccagtgt gccctctgca ccttttatat ctctaaaact gtcccccatg 23700 gatttcatag gtcacccaga agccctcatc atgggatccc tgtgtgagaa gacctcccgg 23760 ggctgcttct ccacagatca ttgaggggag gggtatggca tgctgtgagg cagctctgag 23820 atcattcatg tggtcttaaa ggagagagct ctgtcactgg tctgagagcc tggtgtcctc 23880 aggcccagtg tgagtcatcc tgctccccca accccccccc ccccccggca aatatgcttg 23940 gcttctgagg ttgactattc ccacagactt taggaaacaa ccctattcct cacccctgtc 24000 ctccacctct caccttccac ccctgtgctg gtgtttttct cctccctcgt ctgcagttta 24060 gaaagctttg ttccaaacat ccagagcaca cgcctccctg cctgcccagc accctttagc 24120 acaatgggag gattgataaa tgccatgcca gggtcatgcc aagatgagaa gcaacatgaa 24180 agtaatagat aaaaggggaa gaccagacag tgtgtgggga gcagggcaag ccctccccac 24240 atgcccagca gcctggctct gggataggca gttacccagc tagtactgct ctgtggcttc 24300 tagcatgggc ctttttgcta gcttggttcc atttgcatgg cagattgagc catattggtt 24360 gctgttcaga gtcttctcca atggagatcc tggttgcctg gctcggtggt aaaaatttca 24420 actggctata aatcgtggct agtattccag cactaacctt aagcatagca aagagccaat 24480 ggttgcactc cggttgaatt ttgaaggggt ttggcagtcc tacctgattc cctatgtttt 24540 tccagtctct agcagatcac cccaaacagg gagtgtctct gagcagccta caacagccca 24600 tgggctcaga cctacccacc ctctctctat tcactagtct tcctagctat gggacacaag 24660 acaaatacgg ggctaatggg atgggtttcc cctcctttta tttcctatga ctggttcatc 24720 tatacctcct ctgggattca atctcctcct cagcaacatc atctcccctg cccctccctg 24780 cctccctgtg aatttaaaaa gtgcatatgt gtgtgtgtgt gtgtgtgtgt gtgtgtgtgt 24840 gtgtgtgtgt ggtgtttgta caggtgtgca tgtgggaaca tgtgtgtata gattcaatat 24900 tgggtgtctt cctctgttgg tctttactac tttttagaga caaactctcc tgaatgcact 24960 gtacacagat tggatagagt gcttgtttgt cactaagctt aggaaagcca cttgactccc 25020 cacccccgcc ccaccccacc ccacccccct ctcagtgtca aggttataaa taccttccac 25080 catgtctagc tttgtctggg tgctgggcat ttgaactcgg gtccttatgc ttggactgca 25140 tctccccatt ccccattttg cttttttttt tttttagtgc tgtataacta atatgcattt 25200 tcttgtatca ttaagtattc tgatatctta atgactgtgt gggaatccct gattcccatg 25260 atgcatttca ctcgtccctg ttggaccatg gggctgtttc caggagccag gaagaagccc 25320 atctccacag gcttactgaa tttgtctcat tgtttcatgc gtgcagcctc ttaacagcat 25380 catggctgac tccagaactc ctcactcctt ctgtgagaca gcactggcag gcacctttgg 25440 taaaattatc agctggtgtc ttacattgta tccttctgaa caatggagcc tggccagcgg 25500 tctttgcttg gctatgctct tgctgctttg gggtcttttc ttctagagga atcaagagtg 25560 aagggtagca ggcgggcaag atggctgtgg gagaagatac gctatttatc ctgaggacaa 25620 ggaaagaaaa agggacgagg aggaaaggag ggggaatgga tttctgttct ctccagttcg 25680 ggcagaggtt cgagctctcc cctgctctgc cgcttccgcc tgttccttgt ttcatttcac 25740 tctatggagt ggcttccaat acctcagcat ctcagcatgt gtgcctgatc tcatcttgct 25800 tgtctatgtc agtctgtcta tcctgaggga ggaaaagagc tgaagagctc cgattgtgtt 25860 actcatcacc tacatggaag agcacagccc aactgcttgc ctgcacagtg atacatgggc 25920 gtgtggccca gtgtgaggct ctcttctcac tcatgtggcc aaggccctgc aactatgggc 25980 agggatggat ccttctgact tctctggctt gtctttagca taccacacca atagctaaaa 26040 tccaagctca gacctcttaa cttgttgtct agacacatcc ccttggccgt agggttggat 26100 gttggcggag ggcacagtgt caacacaggt ttgggagtgg gaagtacccc cagacctctg 26160 ttttccattc cttcagttcc aaagtccaag ggcaggcttt tcagctggtg ccagaagccc 26220 aggggccaga gacatgcagt ggactcaagc cagggggaca caggccaatg taaatcactc 26280 caggccccaa actatagggg cttgttttgt tatgccctac ctgtgagtct ttggtacttt 26340 cctcctgcct gcatttggat agcaccatgc cctgccctgc catcttcctc agggcctggg 26400 tgaagtacaa agacaaaagt cttaaatctc tttgccacac agtcactcca cccagccctc 26460 tactagaatg agaagaaggc caacgtgtcc tctaaatacc taaaggcctc cactctgtat 26520 catgctgctg attcccaggg cccagaggag aggccacttg tatttgaaag cagaagaggc 26580 agttgctggg gaattagttc taatgctttg tcttaattag gttcccaaaa gttgcacttc 26640 cagacactga gggtccctac ccagtcattc tgtaaggcaa ttctttattg aattggaggg 26700 aaacagagag ttttagatgg cacgggcaag ggactgatgg ggggtgaaga acagacagaa 26760 tcgccatgaa ggggagcaga gaggtcagaa ttaaaggccc accaccatgt aagaatccgg 26820 gaaagaggcc ccaggggcca ttcccttgct tgggtctagg gaaccaaaag caaaaaaaaa 26880 aaaaaaaaaa agagatttag aaaatgttaa ctcatgggct ggcgagatgg ctcagtggtt 26940 aagagcgccg actgctcttc cgaaggtccc gagttcaaat cccagcaacc acatggtggc 27000 tcacaaccat ctgtaacgaa atctgatgcc ctcttctgga gtatctgagg acagctacag 27060 tgtacttaaa tgtaataaat aaatcttaaa aaaaaaaaaa aaaagaatct tacacagttg 27120 cactggagga aaaaaaaaaa gaaaaaaaga aaacgttatc tcaggaatac tggaggggag 27180 tgtgtgcgag cctcagtgag gtttagaagt gcccagccat tgagttagtt aggcatatta 27240 taaattaagc tggcatgtgc gtgcctttca ttcatgaatg cagagagctc taacgggtgg 27300 ctaaaagtgt gacctgctaa gatgtcctca gtgcaaatta acagttcacc actacaggca 27360 gttgctgcta tggcttacca cccagttagc ttgggtgtgg ggatcttcag ccagagggca 27420 gcatatggat cttaaagtga cacagaatat gtccttgtca cagggcgaga ggttttccag 27480 ctagagtcac aatgcgtaca aaggtacaga gaaaggaggc aagtgggctg gaaacgtctc 27540 aggagataag gagtggctga agctggaata gctaagggtt atgtgtgaaa gagagcctgg 27600 cagagaacaa gggagcagcc agaactcggt gtccaaagcg cctggagaaa aattttggag 27660 cagatctgat ttagatttag aagggagact ttggaagcct gaggaatgtg gggcagataa 27720 tcagctagga ggtccaaggt tacaaagata agatttgggt aagagtcact atctctcccc 27780 agaggcgata tgggaaccag gtttatagcg aagtccaccc tgagggacag cacacccgat 27840 atggacagtg tgagtgtcag ggagtgtgtg ggtcggagcc ccagttctgt caaccagccc 27900 cgagaccact cagcctccct gcgacccaca gctttagctg ggtggcagac gattgtgggc 27960 atacaagacg gcagccaact cacgtgacaa tggtggcctc tcctggagaa ggatgactgg 28020 gtctcaggag gagagggtct ctggcctgtt tcacgagaca gctggactaa gagggagctg 28080 agttgcagtc taaccaacct cctctcttcc cattttgttt tacttttttc aagcagcagg 28140 taagctgagc acacaaaagc aaagacgtag gctctaggag ctaaagcagg gatgaggcag 28200 aaaaaaaaat agatgaactc tatccaaaaa catgcatcaa aaatataaac atggctgagg 28260 ctagagccca ggggcaggaa cacattctta gagtgtgctt gggttcaatc caaagcttca 28320 aattccttag caccacacaa aaatttaaaa gtactatttg tttttaatct gatccagtaa 28380 tcctacctct agaggtctgt cctgtagaga gtctgagatg gatggcttgt taacggctat 28440 gtttgatctg acccacatca tttttaaaat atttaaatgt tcaaaaacca caagttttat 28500 atatatgtta ttatataaat atatttttat attatattat tttattctat ttattctata 28560 ttatatttat tattatata tattatatat aataagtctg actatacaca tatattcaga 28620 cttatgataa ctcttggtct ccatttgtct tagttgcttt tatttctaag ataagacacc 28680 atgaccaaag taacttataa aagacagtat ttaattttgg gctcatggtt ccatagggtt 28740 agaatccatg agcatcgtga cagcatgaag catgacagga ggcaggcagg caggcaggca 28800 ttcaggcagg caggcaggag gcaggcaagc aagcaagcag gcatggtact gaagcagcag 28860 ctgagagctc atgtctcaat ctgcaagcat gaggaagaaa gagagctgac tggagatggg 28920 tgaggccttt ttgaaacctc caaacaactc cagggacaca ccccctccaa caaagccaca 28980 gcccttaatc cttcccacac agttccactc actggggacc aagtattcaa gctcttgggc 29040 ctgaggggac attctcatta tccacaacac acctactggg cagcaggtgc caccactgag 29100 cagcaatggc cttctctgtc cagctcatgc ttccatttcc tcaacagcag cccctgactt 29160 ctcttagctg tctaatagag gcttgggggc acagttattg tccttgcatt atagatttcc 29220 cccttaaata tttttaattg acaaattaaa aaccatgtac ttatgatgga gtattatagt 29280 gtgaaatggc taaattgaga ctgttaacat acgcactata tcacggacta aattttttgt 29340 gctgagagca cagagccagc tcagccattt tcaaacagac atgttgttag ttattgtcct 29400 ttctcaaatg ttgtagcttt tctcagacta gggacatatc acactgttca ggatctccat 29460 caaaggcaaa gaaactgagg gtccatgtgg aaaccccacg tctaagggag caagagaaac 29520 ccatcatttc tttgtggaaa tggtggttgt gttgtaggtt ccacagcaag tccctgggcc 29580 ctcattcatt tattatttat tattgtgcat tattgattat gatgctgaca catgagccac 29640 agaagtttca tgggcatttt gaatcttccc tccttgatta tctctcggat ggcatggtct 29700 tccccctccc gccgtgcttc tttttgaata cacaccaaca gcactgatga gctgtgtttg 29760 tggctcagtg tagactgctg tgcagggtgt atccccagca gcacacaaga ccaaaaaaca 29820 acgcgactgt gacaacagcc aaatagaaac caagtccttt ctgctatgtt tgggtaagga 29880 ggccacccac gttcacagcc cagacttctg acccatttca tatttcttac gggaagtatt 29940 ccatttttgt tatcaaaaca agaattatca cggcttagca gtaagcctct tggtctgggc 30000 tggagatgca gcgcagcagc cgagcacttg ccctaggcta gcatgcacaa gcgcctggtt 30060 tcaatctcca gtccatggag aaaaagcaga aggaagtctt ggcttgagag aggctttagt 30120 gatggatcac gggactgaaa tttacctctg cccatcttgg cagctagcag gacaggtgca 30180 gtgggccgtg ggtttctgga agagaccata ggaagaggaa accactttct ctaggctaag 30240 ccatcaaaag attctgtccc ttaggacagc aactaggtgt ccagctctac acacactaag 30300 cacactggct tcttggtcct tggtttggaa ggtcaatctc tcctcctcct cctcctcctc 30360 ttcctcctcc tccctttttt tgagacagat tctcatgtag cccaagttgc cttcaaactc 30420 gctatgtgtc tcaggacgag cttggactcc ggatcctgct gtctccacgt cttaagtgct 30480 ggttttatag gcatgtacgg ccacacttag tttatctagt gctggagacg gaatccaggg 30540 ctttgtgcag gctagataag cactctaccc ccttgctaca tccccagccc tcctctcatt 30600 catttttctc attattttag aggcaaaata atacttcact aaggtgtagt ttatagacaa 30660 tttaattcac ctattctaag tatgcaaccc aaaaacaatt tttttaactt atttatttta 30720 ccattttaaa aaatttgcag tgttctgcct gcatgtataa ctacctacca gaagagggca 30780 ccaaatctca ctatagatgg ttatgagcca ccatgtggtt gctaggattt gaactcagga 30840 cctctggaag agtagccagt gctcttaacc tctgagccat ctctccaggc cctaagattg 30900 cttatttatt atgtgtttgt gggttggaca catgaaggta ccatgctata caggtgaagg 30960 gcaacttgca ggagtcaatt ctttgcttta ctgtgttggt ggtagggata gaactcagat 31020 cactaggcaa gtgcctttta gaatatttca tcaaccccaa aagatttctt cttatggatg 31080 ttcccattcc tactataggc aaccaaccaa ccttttcctg gatacttctc ataactgaat 31140 tcacaggata catggtcttt ttcatctggc ttctttcact gaacatggtt tgagctgtct 31200 ccgtgttcaa atcagaatca gtactttgct ccttcttacg gtagaaatgg atcccattgt 31260 atagatcact tacatcgaga cagcaagcct ttagtcacct gtctgtggat gttggagtcc 31320 tttcccctct tgggctgata gataatattg ctataaacat ccacatacag acatttgtgg 31380 ggatacatgt ttttgtttgt cttgtgtgga tacctagatg cctcaatgcc aggttctata 31440 gtgactctct gagaaactgt taagctgtct cccatctccc aatagggcta taccactgcc 31500 ttcattccca ccagcacttc ctgagcatcc agcttctctg tcctgactga ccccgcctac 31560 cttccatgat tacagtacat ccaagaagaa gaagataact tcttagggtc acattagaga 31620 gtgacagggt caaggtttgg aactcaagtt accctgactc tgcaatccct ccttcagctt 31680 tctggttagt ggatggaatc tgtaaaaggg agggttggag actggggctg ctgacctggg 31740 gctgcagggt aggactcaga ttcacaggaa tgccttctct ctggtgccct cccctggcct 31800 agcttgtatt tctctcaggc tgccatgtcc ctggcctcac gtttccttct gactgggcct 31860 gggcctgggc ctgttcttct gcctcagagc caggagcttt ccctctttct tccaacttca 31920 agatctattt gcagtctaag aatgtatcta agaagggcac cccgtctgtt cgtttaactc 31980 atataggatc agaggcaaga taccctgttg atgccagcct gaggctagct agcggtatga 32040 ggaaattggc ttctgggagg tgtgatggag tgtgtctggc aagcaggcag cagtgaagcc 32100 tcaggggtcc tggacagctc actctctacc aacttgtgct cactctttta gaatgatcat 32160 cctggtcccg gggagtatcc cagccagata tggtggctca tacctataat cccagtattt 32220 taaaggcaga ggcaggagga ttgtaaattt gaggttagcc tttaaagact ctgaaaacaa 32280 acaaacaaac aaacaaacaa acaagccaaa aaggtctata agagaacacc caagcaaact 32340 tcctgtccaa acaggaaaat ggaggagaat gtgcatttat tgagtacctg agtctccctc 32400 atctatacat ttcttccaat ccccgtactc actgacaaac gctgcacctt ccctctgagg 32460 aagtagattc aggctgtagt atttgtctca actcacatgt tcagtagttc agccaacagg 32520 aggatgcctc ctggctctgt ttctgcccac cacacacagt gtaccaaagt gtggggtaaa 32580 acatcaggtt agtatgagtt caagtcctga ctcggcctca tctttgctgt gtgacatgga 32640 gaaagtgagt aaagcggtct gggccccggc tgttcatctg taacataagc ctgtctcagt 32700 ccacaccccg aacgaagata gaaagagaag gctgtggcat ggttggacat ctagactttg 32760 gctgtcgggt ctttgcaccc aggctcagct gactagggat cttaggcctg ttgcttttca 32820 gtcccctcca aagccttgac tttctgtcca actggatctg tccccacctg ctggcaggag 32880 cctgagaagc tctgagccca gagataattt tattcttgtg agctgatgcg agctttgctt 32940 ctgcccgtcc ccatccttgg caagcaagct ctaactggat aaaagcaaaa aaaaaaaaaa 33000 aaaaaaaaaa aaaaaaaaaa aaaaaggcct gtttctgtct cctcttccct ttcctgatgt 33060 atcccagagc tttgctcatc ttccaaggcc aagtagcctt ccagcccacc ccaggcctgg 33120 ctggcccgct agctctcaag attctgttct ggaggcagcg gcagtagcag caagcatgga 33180 agctctgccc aggtcagaac tggattctct gaagacaact caggtaggct tgactagact 33240 gtggtcatct ttcttgcaga ggaagcacta tggacaaccc gggcggatgg gagagtgcgc 33300 ctgcgcctgg aacctttctg tactcagcgt ccctataccg tgcaccagat gttctacgag 33360 gccctagata agtacgggaa cctcagtgct ctgggcttca agcgcaagga caagtgggag 33420 cgtatctctt actaccagta ctacctgatt gcacgcaaag tagccaaagg cttcttgaag 33480 gtaggccccc atcactctac atcccttccc ctccccctct caacccctga cttactttct 33540 agctttagct ctcaaaggcc tcctccccaa ttccccatca ggcaaggtcc ctagaaacca 33600 ccattgtaaa gtctgacacc accagatggg ctaacactat atctgggccc tgtgctaaga 33660 agtttacatt tatggagcct ttatcacagg ccaccttcca gcaaggctgg gaaacctagt 33720 agttagggct ccctagaaaa acatagaaag gatcaacagg gctgccagaa actgactccc 33780 caggaggttc ttcacctggt agaatctgag gtggacctgg tggggtctct gcagtctagt 33840 ggagagcaag acaaaggtgg gaggtgaggg ggaggccctt tctacccttc tgtgtagctg 33900 acctgcctat ctgcctttgg cctggagctc ggcctagagc gtgcccacag cgtggcgatc 33960 cttggcttca actctccaga atggttcttc tctgcagtgg gcacagtgtt cgcagggtga 34020 gtgactaagc agtggttggt gatgggcaga gagggttatt atctccttag cgagccctct 34080 gcctctcccc agcatcccgt atcttcccta gggccttctg ggcctgcttt tccccatccc 34140 tgcctgttga gatgactaag gaccctacct taagacctct gaagttcccc gaggatggag 34200 gatgtgtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg tgagagagag 34260 agagagagag agagagagag agagagagag agagaggttc ctctgctcct gatatgattt 34320 tcttgggcat gtaggggcat tgtcactggc atctacacca ccagctcccc ggaggcctgc 34380 cagtacatct ctcatgactg ccgagccaat gtcatcgtgg ttgacacaca gaagcagctg 34440 gaaaagatcc tgaaggtttg gtgggaagga gagtgagcat gtggttagat gtggttagaa 34500 gctggacatc taccctagag tgggacggat actccacctg ccacccagca atcttattcc 34560 tacagcccat gtgaacatgc aacatgcatg tatgcacatg cacctgtgcg gggacacaca 34620 catactcaca tacagaaaga gaggggcaga caggacaggg acaggggagg aagttaagtg 34680 ggtaatagca cttttcactt atacgtctgg ctagcccatt atctaacatg gcttttcctc 34740 tggacagggt cttacctaaa gggagggatg tggacattgg gcctctctgg cttactttac 34800 ttgctggaaa tgtgccctag accacagcta ctcattggct cctgagggtc aggtgctcag 34860 ccttcttctg gagaacctgg gactccagag agaccatggt agatgcctcc cctcatttct 34920 gttggagaag tctcttaagc caagaagaag caagctggaa cttgtagttc tcacttggct 34980 gtagaaatcc tccgtagggc aggcccaggc catgggtgag acccccagca ggacctccta 35040 aacagaggag ggttccctca tggtttaaca tagctcactc aggtgaggct gtgagtgctg 35100 actaagaatc actagctcac agagagacct ccaggaagct tccggcgtcc aggcctgctg 35160 aggtcacact agtgatggga agaagccaaa accggctcct ccctgagaga gactgggaag 35220 aaatgcagag ctgggagcca ggagagctgt aggagaggag ctgaggggga cccagacccc 35280 caacaggtct aagcgggtac ctaaatcatc aacttaagaa acaatttctg aaacagtagt 35340 gttgacctgg cagggtgtga ctacacaagc cctcctgaac ttgtcagtga agctgaataa 35400 ggtcctcaac ctcgcatctc gcacttggcc accctgtact agattcagac tctgtgggag 35460 tttggtcagt ccaagacctg tgaccataag cttttatacc agtttagagt ggttttggaa 35520 actccattcc ccctcccttc tgctttactg ggaagagttt ctccagtggc tcccttgaat 35580 agttgaatag tgaggaattg tcctgtctct ttatggggtc cttttttttt tttttttttg 35640 gtttttttca agacagggtt tctctgtata gccctggctg tcctggagct cactttgtag 35700 accaggctgg cctccaactc agaaatccga ctgcctctgc ctcctgagtg ctgggattaa 35760 aggtgtgcgc caccacgccc agctatgggg tccttttttg agcccggctt tcagtatatc 35820 actatgagga tctccatttg tccagcctgg ataaatgtgt aggattggga atggacggat 35880 aggtttagat tgtgtggccc caaccagtac tctgaaacaa ggaggactga gctctctctt 35940 caggcatcct tctcctcttg tcaaaactcg gtctgtgggc caactggtca accttctaga 36000 accatgggac ctgagtttcc agagaggccg tggaaagtgt cttcccttgc ctccactgga 36060 atactcatca gggatatata tctgttcctg gcatttccaa agcaccctga gactttttat 36120 gaccttgagg tactcagata acctagcagc ctcagaagca ctgaggtcat taggtcgttt 36180 tctgagcctc actcattatg gcctaactca gaacagagct aggctcccca aggcttgctt 36240 aggcctgaag ccttgatggg tcaggaacca atctagcata gaactgaggc ctcatgcact 36300 ttcctcatca ctcaggttag caggcctctg gtgttgtcct tgttcagctc acgtttaggt 36360 agtcatgatg atgagacttc atgggtgggg cttctgatac tcctaggaga ctcaatctca 36420 caaggaacac tctgatcctc tagctcttaa atctttccag cacaaagaac tacaggcaac 36480 tatggaatgc tgggagctgg agagttaatc ttccccaggg atgagcacac caactctcca 36540 ataccaaata gtcagccctg aaaacataat atacaagtaa cattatagag actgagtatt 36600 tatgtattta ggagtgtgtg tgtaacaata attaatgaaa agagaggtca tgaatttgaa 36660 cgagagctag ggtctctggg agtgtttggt gagaggaaag gggatagggc aaatgatgta 36720 attgtaatct caagagaggg gtggggtttt aaaagaccca atccagcttg gccagggaga 36780 ttgctcggtc agtgagtaaa gcatcttcca ggtgtgcatg agaccctgag tttgcatcag 36840 caacacacac atgaaaagcc agacacagca gcaagtgcct gtaaccaagg gtgcaagaga 36900 aggaagaaaa cagacccctg gagctcattg gccaagctta ctgaaaggcc tcagataaaa 36960 aggtcagaga gtggttgagg aaagacatct gctgtcaacc tgtggcatgg cacacttgtg 37020 caggtgcaca cacatgcaca gagggcagca gagaatggtt aggtcccatc ccctgaaatg 37080 gcagtgccac tcacaatgtc ctaaccaaca gcctgggctc agctcgggct agactgagaa 37140 ggatcgctgg caggcagata gaagttgaca ctcagcaatc ctcaggaacc tgcctggcag 37200 gaagagcctc cccttccctg cctctggcaa atggggcatc tagctccctg ttaacatata 37260 catgtagcac atatcaaagc tccaagctaa tctccaggct ttctcctcca tattaacatt 37320 agcattacct gtgatgtatg tgaaatccat tcggctcttc cagcctgcaa acttcctctt 37380 ccctctgccc taaatttcca gcctacttca gattcttctc cctcccttcc actgtcacac 37440 tgttctcttt aaaactggca aacacacctg ccccccctcc cccaggtaga catggcagct 37500 ttggactccc tgaaatgcct ctggtttctc tctccccaac ctgacagcca tgagacagtt 37560 tcatatttac aacaaacctc caccccaccc atgtaatggc ctagttcagt ggtttcgctg 37620 agccctcaca agaaggaaaa ccaactcgag gccctgtggc cagtggtatg ccgtctcttg 37680 acccctccca tgcttatatc tggatctggg gggaggggga agctggacag cttaaaaagc 37740 cgccacaagc ttctgtcagg gaaacaaacc atccctagct tacagctgaa gaagatgaag 37800 tctacctcta ggacctcagt aaaatgcctg ccttacgttt aaaacacaat ttgtgttatg 37860 aaaattctca aagaaataaa agcagagact gatatgtttt ggaacgcagc ggcatgatgt 37920 ggtgggaagg ggtgccttag tggccctgtg ctgaagcatc ccccaccccc accccaaggt 37980 accagccata tgacaggtct ggtataaaat gaagtttatt tgagggcctg agaagtgtgg 38040 ggggggtggt gagaaggaat gtgggaaaaa aagaggacag agagggacag aaagagagag 38100 gagagagaaa gagggaagag gttggctggg aacatgtgga gagacatggg gggggagcag 38160 tgcgtgtagg gagagaaaga caaggatgag agtgtaaggc cagaggagtt aggggagcca 38220 aacagtcctt ttatagcaag ccaggctcct acctggctag gtagctgttg ggcggagcct 38280 agaagaaatg ctaacagata atccctgtat catcaggttt cacagttatc gtggtcctcc 38340 tgtgcctcca cagtgctgag atcacaggca caggctccac tcttgcctgt ggtgttcact 38400 cctttcggat tgctcccaag tactgaagaa tcatcttttc tcttttccac atgaatttgt 38460 taaataatta caatattcac ttgatcccaa tgtcaaaatt atatgttaca cttgaacttt 38520 cccttccctt tgttcctatg cttgctctct ctctttgcct gtagaaagtc agtataattg 38580 gctttaaaca tcctgagtcc cttgtggcaa acataagcaa atgcctcttg cttaggtgtg 38640 tgtgtgcgtg cgtgtgtgtg tgtgtgtgtc atgtgtgcac aagtgtgtgt gtcatgtgtg 38700 cacaagtgtg tgtatatgtg tgtcatgtgt gcgagagtat gcacacatgc atgcattata 38760 cacatgtact cacactgtat tcctttctca ggtgaagata ccgtgctaca aacacagtgt 38820 cttcctcact tagcaatact tcctacgagc cattctgtat tgcactttag aattcccttt 38880 gtacctctgt ctctttgtca tctgcactaa cctaatattc tactgtgtga cttgtttaac 38940 caggcactag agccccactg tcactgcacc caacttccca taatggggaa tgctggacag 39000 agatgtccca ctggagtcga gggtcagaaa caggagaact ttgtcacaaa ctaacatagg 39060 tagatgaggc tctttcctag atgtgaccat tttcttgcta aaaccaaatt tgttttccca 39120 tcaatgcaga tctggaaaga cttgccacat ctaaaggcag tggtgatata ccaagaaccc 39180 cctccaaaga agatggctaa cgtgtacacg gtatgaatag cgggccctgg gtgatgagtg 39240 gggtctgtct atgctaccca gctatcccag actgacttct ctgccctacc ttgccacaga 39300 tggaggagct catagaactg gggcaagaag tgcctgagga ggccctagat gccatcatcg 39360 atacccagca acccaaccag tgctgtgtgc tcgtctacac atcaggcacc acgggaaacc 39420 ccaagggtgt gatgctgagt caagacaatg taggacagac acccagatgg atgggtggga 39480 tggggggact ctgggctgtg gtcctgtaga cagacaccca gatgtttggg tgggatgggg 39540 gaactctggg ttgtggtact gcctcagtag cctcttgggg aagcctctct ggctgttaag 39600 tttatggggc tagatgagtt ggaacccctt tgtggtgtag acagagccct atagagggaa 39660 aagggaaggg aacagatggg aggcctgtcc ctggagctct ccagagcctg tggaaatgcc 39720 catgagggca cctacctact ctgagaacct gtaaagaggt agcttaagcc cctaaggaga 39780 cctcactcta catacttcaa ggactgggtg tggacctgct ggggccattt tccttcccct 39840 cagatcacat ggacagcacg gtatggcagt caggctgggg acatccagcc agcagaagtg 39900 cagcaggagg tagtggtcag ctacttgcct ctcagccaca tcgctgccca gatctacgac 39960 ctgtggacag gcatccagtg gggagcccag gtctgctttg cagatccgga cgccctgaag 40020 gtcagtctgt cttcctgact tcctgtcctg tgtgagcgag ggcgcactca aatgtgtgct 40080 tgcatctatg ggccctgtgt gcatccattg gttgctgact ccagtgcaca tagtgctcag 40140 aattctttta tttatttatt tattttttaa tttctgcagt ttttcgagac agggtttctc 40200 tgtgtagctg tctggaactc actctgtaga ccaggctggc ctcgaactca gaaatcccct 40260 gcctctgcct cccaagtgct gggactaaag gcgtgtgcca ccactgcttc agcagctatt 40320 tgcacaccct gacttagatg accgaacaat tagggaaagt cttataaact gccttcttca 40380 gtctcaaaca ctataaggct actgcttcct cagagtatga ccacaagatg atcagagggt 40440 aggcatagct ccctgtaccc catctcataa aatatatgta ctctttgcat ctttattcta 40500 accgtgagga gaggttccca gaaaccactt agctgtcatc ctccaacata gtgccttggt 40560 agcctggaca gtcatatgtt tatttctaaa gcaaactctg ctggggaagg aggaggccaa 40620 atgtactagt ttggctattg ttgttgttgt tgttgttgtt gttgttgttg tgtgttatgt 40680 gttcatgtgc tcacctgcgt gcaagccatg tgaaggctag aagtcaacag aggttgcttt 40740 cttctataat tctccatctt taaattacaa aaattaaaga ttaaataaat acttaattta 40800 tgtttgcatg acataaatgg agaccagaga gggcacaaga tccctggggc tggagtggca 40860 ggtagttgtg ggactcctgg tgggtgggtg ggaaaactga actctggtcc tctggaagag 40920 cagcaagtac tcttaacggc tgagtcatct ctccaacctc gtctgtctta ttttttgaga 40980 caggctccct gctctctgta ataaatcaag tgaaattagt tttgataatt ttaaattcaa 41040 ccttacagat atattttact gatatatttt aaaaacattt actttatgtg tatgaatttt 41100 ttcctgcatg catatatgtg tacacccatg tgcctggtac cctcagagag cagaagacag 41160 acgaggtcat tggatctgct agaactggag tcatagctag gacttgtgag ctgccatgtg 41220 gctgctgaga actgaacctg gctcctttgt aagtgcaaca agcattgtaa accacagaac 41280 catctttcca gatcttacaa ttaatgtttt taaatgaggg ttttttacat ttattattat 41340 ttttttcttt tttcaagaca gggtttctct gtgtagccct ggctgtcctg gaactcactc 41400 tgtagaccag gctggcctcg aactcagaaa tccacctgcc tctgcctccc aagtgctggg 41460 attaaaggtg tgcagcaccg cccagctaca tttatttttt aataaataca cctatctatg 41520 ggtataagga cgaatgttta tagattattg tgggattgtg ctactttagt acattagggg 41580 ccatagcttc tcctctaatt gatagcacta ctcatactga gcagttagct aggtttccag 41640 gatcaagcat gtttacaccc tgtacatggc tctaagtccg gttatagag ttttggttac 41700 catcaaacga tacaagcttc tcctgaacct tagggttatg atgctgtgat gactgcagag 41760 atggttccta ggcagcatgg ctggttggga cctgttgctt gcttccctcc tttggaagct 41820 tgcattggtg cctttgatac catgaaagct ggctgatagc agaggagggg agggggcatt 41880 caggttagtt tcagttcagg ggcttctgag ccctctttct gaggttgaga tgtcttcagc 41940 aatggggatt tactttctat ctcggggtgg aggtcagcca agggcaatag caataggctg 42000 tgtttgagga gtctattaga cagccctggc taacagcgca aaagatgggg gttttgttag 42060 attgtctttg gcttttggat ggagcacttc agatgaggaa agtttattag aactatacat 42120 gtttattcat atggggaatt gtgcgttgtg ggtttctttt aatggtgtga ttccttgtgg 42180 cctttcaggc gtccttattg ttatttgact catcttctgt gtgtacctcc cttccacttc 42240 cctgaagagc cccccttccc atctctgcta tcagctaatg tgtgccctct tattctctct 42300 ctgtgctctc tgtccccctg tcctggcaat gatcctgctt tactctcctg ccttctgtag 42360 taactgcagg ctgtatgctc acatctgaag acttggagcc acgagcctcc aatgggagag 42420 agcatgtgat gtttgtcttt ctgggtctgt gttaccacac tcagtatgat ctttcctagt 42480 tccatccctt tgccttccaa ctccatgagt tcatttttct cgacaacgga atgacagtgt 42540 ctcattatct gagtgtaggc tgtttatatg aggcaaggct ccctattgag cccagagttc 42600 cctatttggg gcatactttt cagttattga gcccctgggc tctactctcc aacatggcgt 42660 ttcaggcgca tgtgcagtga ccgtctcttt gagtgggtgc tgaattcggg ttcccatact 42720 ttcttgactg agccattatc tgagcccctt tgtgttgctt ttaaataatt ttaaaattac 42780 atttatttgt ttttgaggca aggatgtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg 42840 tgtgtgtgta gaggagagag acattggtta gctggctgtc ctagaactta ctctgtagac 42900 caggctggcc ttaaactcat agatatccac ctacctctgt ctctgtctcc catgcactgg 42960 gattaaaggt gtgaagcact gcatccttta tttatttatt tatttatttg tttgtttgtt 43020 tgtttgttta cttattcgta tggaatacag cagaggtctt atgtgtttat agtgcacagt 43080 gtagggtca aaggacaact gtcaggggtc tagtgtgtcc tttcaccatg tggcttctga 43140 gaatgaactc atgttgtcag gcttggccat agtcccctct tacccactga actatcctgc 43200 tggcccttgt tttcatttgt ttgtttgttt gtttgttttt tgttttattt gtctgtttgt 43260 ttatcttgag atgggctctc attagatggc tcaggctggc tttgaacttg ctatgtgccc 43320 agcctgggcc tcaaattcac catcctcctg cttctgcctc ctaagtgctg aaattgcaaa 43380 catacaccac tatacctggc aaaatgaggt tatcatgttt gaattagaca aatgcagaaa 43440 ggtctgaggg atcaagttgt ttctcgagca tcaggctaca gtggttggca aaggacacca 43500 aagttctgtt agcagatgtg gggatgcagc agttgaacca cagccactgg catgcgcaca 43560 aactggacac aagtacaaat gtacaagccc acaggagtat ctgtgtgtag cattagcaat 43620 tatactgaga ggctgtgtct ccaggcctgg taccacctct gcttctctct ggccttgagc 43680 aggggaccct ggtaaacaca ctgcgggagg tggagcccac atcccacatg ggagtgccgc 43740 gtgtgtggga gaagatcatg gaaaggatcc aggaggtggc cgctcagtct ggcttcatcc 43800 gccgcaagat gctgctgtgg gccatgtcag tcaccttgga acagaacctt acctgcccta 43860 gcaagtaagt tggtggcagg aggcctgggg aggcacaggg ctagttcaca tacagaacga 43920 gcaagaatct gataggtatc ctggttccta aagacttcag tctgaggaga ccttgtgcct 43980 gtttacaact gggcttgtac tgtgataccc attgtagaca taatggggac agctggcatt 44040 caggggcttt gtctgggagg acttatttgc ttcttccgag taattcatct tactaaggat 44100 agatagctct tgccagctgc ttgcccgcag tagtggcttt agaagggtta tcttagttaa 44160 gacctctatt gctgcgaaga gtcaccgtaa ccacagccac ttttataaag gaaaacattt 44220 aactggggct gacttacagt ttaagaagtt tagactgagc caggcatggt ggcgcatgcc 44280 tttaatccca gcacttggga ggcagaggca ggcggatttc tgagttcaag gccagcctgg 44340 tctacagagt gagttcccag gacagccatg actacacaga gaaaccctgt ctcaaaaaaa 44400 ccaaaaaaaa aaaagtataa aaagaagttt agactgttat cataatggtg ggaagcatgg 44460 catcgtgcag gcagacatgg tgctggagaa ggaactgaga ggatctatct ggatctgcag 44520 gcattggaag gagactgcca cattaggcct ggcttgaaca tttgaaacct caaagccagc 44580 ctccagtgac acattttctc caacaaggcc acacctccta atactgtcac ttctatgcac 44640 caagcattca cacacatgag tcaacagggg ccactctctc tctctttttt ttaagtacta 44700 tgtatttatt tttatttata catgtacacc gtggctgtct tcagacacac cagaagaagg 44760 catcagatct cattacagat ggctgtgagc caccatgtgg ttgttgggaa ttgaactcag 44820 gacttctgga agagcagtca gtgctcttaa ccactgagcc atctctccag cccaacaggg 44880 gccattccta ttcaaaccag cacagggggc ttcagtgata caggcttagc cactccagct 44940 ttcccagcag cccattggac tcctcagacc tctctctagg cccttctgag tgcctattat 45000 cattactttt tgttttctga gacaatctta atgtgtagcc cgggctactt taaggatctg 45060 gctcctcctt ccacggcctt ccaagtgttg gaattatagg tatagaccgt cacatccagc 45120 tcatgactcc tttccaagtt cccacacgct gctgtgcaga tatccacagt agtgatctgg 45180 cctcgtttca acataaaaat gcctttcaag ttgaagattc agcttggagt tagagcactt 45240 gcctacaagc tccaagctct aggctcggaa cttgtgactt caaagagggg aggggtggag 45300 gggaggatgt tctattatag tcgtgccagc tctgtgagag atgtggacat atctgttcac 45360 ctatggcttc attaacacac atgaaaagtg accatgggag tgtagctttg taatcccagc 45420 attcaggagg ctgaggcctg aggatcaaga gttaaggtct gtctgcttat gctatggagc 45480 aagttcaagg cccaagtaaa ttcatgacac tgtctcataa taaataaata aaaaggggtc 45540 tggagaaatg gctcagcttg tttgacttca tgattcatga cacatctaca tgcattcagg 45600 agcactcaca cttacatttg cacaaattca tgttacatgc aaggccaata tccaaaatgt 45660 ttacaaaggg tttagcatga acaacaattc atccaaacgg acagcttggg cggccatgct 45720 gggtgcatac tcacgagtgt ctttgttctt cctcctgtgg atcattctcc tccccgctcc 45780 tctcacagtg acctgaagcc cttcaccagc agactggcgg attacctagt gttagccagg 45840 gtccgtcagg ctctgggctt tgccaagtgt cagaaaaact tctacggagc agctccgatg 45900 acggcagaga cgcagcgctt cttcctgggc ctcaacatcc gcctgtatgc aggctatggc 45960 ctcagcgaga gcacaggccc ccacttcatg tccagcccct acaactatcg gctttacagg 46020 taaggcttga agttgggaca cactgagggc tcagatgtcc tcgatgacac cctttggtgc 46080 cttggggaac ttgttcctct cgctttcccc ttcacactga atccctagcc tcaagtttca 46140 actgccatct tgggcccctt gcttgtcctg atgcatctag agaaacagtc tttgccacag 46200 cgacatgcac acacacatga gcagcaccaa actcgggaat aacacacaca cacacacaca 46260 cgagtgtact tgcatacgct tgtgtgcaca catacaagta aacatgtgca catgcatatt 46320 tacacaccaa aacaaataat aatgctaaca ataatagtag tagtagtggt gatactaaaa 46380 taataatatg tagtagtacc aaaatctgaa aaagacggcc tggagaaata cctaagcagt 46440 taatagcgtg tactgccctc gcagagtttc tagttctatc cccagaaccc atatttggtt 46500 gttctttaac tgcatagatt tctagatcca ggggacccag tgctcttctt tggatgctgc 46560 atgtatctgt gtgcatgcat gagcacacat acaattattt ctaagtctta aaaagcagtt 46620 ctgagggctg gagagatggc tcagtggtta agagcactga ctgctcttcc agaagtcatg 46680 agttcaaatc ccagcaacca catggtggct cacaaccatc tgtattgaga aacaaataaa 46740 aaaaaaacct atgggtggtg tccagattga gcagggacta gagcgagtgg ggcccagagc 46800 gatcgggacc cagaaaggca gggcgggggg gaaaagcaat cctgaggggg tggggcccaa 46860 ccaccaatga agactttgca gactcaggag ctgactcagg actttacctg acctcttctc 46920 ttttatcacc agctccggca gggtggtacc cggctgccgg gtgaagctgg tgaatcagga 46980 tgcagatggc atcggtgaga tctgcctgtg gggccgcacc atcttcatgg gttatctgaa 47040 catggaggac aaaacctgtg aggccattga ttcggaaggc tggctgcaca caggtgacat 47100 gggccgcctg gatgctgacg gcttcctcta catcaccggg cgcctcaaag gtgagcatcc 47160 ctgtcctggg gcacatggga actagaggag caatcggtgc agaagtaagt cctgcttcca 47220 acacggttca tcggcgctgc accctgagcc ttcactacac gtctcttatc agtatcagct 47280 cagggcaagt actgtcacaa ataaacaagg atgatcggag agattagata actagttcag 47340 ggtcaccgag ataacagccc ccaggtggca gttccacttc ctttctggaa cttacatagc 47400 cagaggtact gctcaaccta gttccagctg aaagaggttc tggtccttca gaaatgcacg 47460 gattgggctg tactaagcag taagccaggc agtaaaaccc aggctctaga gcagaaggca 47520 gctgctaggg gaactgtgct ctcttccacc acctagtgtt cacttttata atgacaaatg 47580 aagaaacctg agttccagcc ctgagaggaa aggccctgtt tttaggtcag tgggttctca 47640 acttgggggt caggaccacc ctttcactgg ggtcccaata tcctgcatat gaggtattta 47700 aattattatt tattacataa ttacatctat gtaccttata taacaaaatt acatctacga 47760 agtaacaatg gaagtaattt atggtcagga gtcactacaa catgtattaa agggtcacag 47820 cattaggaag gtattgtttt agatggtatg gtgaaaggta agactccaga taccctgttt 47880 cttcaaaaaa cacagcagtg ctgcccacca aaaagacacc ctttctcgtg gtcctatacg 47940 gtcaacgttg agactatttt gtcccaaggc accagtgtcc ccttgagtag ctttagactg 48000 ggctggtgtc tggttctgtg gattctgtgg cctgatcatc ccctccatgg taccttggaa 48060 gttcctttca tttgcctttc cgtttgaatt tagactcact gtattggtgt atgcaatagg 48120 ttggcttaca tggtgatttt catctcgctg agtctagatt agttttgtga ggttgtacat 48180 ccctcagagt taagtctgcc gaatcctatg tacactgttt ctcccccccc ccaacctccc 48240 cacccgttta cttaggtctt tgacttttct ctgcagaact agtgttctag ttttcagtgt 48300 agggtttttg catccattca ttttaagaat tttattcatg tgctatggtc aataatggca 48360 tttaaatttt tactattcgt tcctgagata taaaatgtaa ttacttgttt tttatatctt 48420 ggtcttgggt caaatgactc ttttgttgtt ctaaggatta aatataggtc tttgagccag 48480 ctaggcgagc tttctaccac tggccagtgg ataggccaga cgttcttttt acttgagaca 48540 gggtcatgct ctgttgtcta ggctggtctc gaattttgaa tcctcgttgc caaatagctg 48600 ggattaccca gcaggccctg cttcaaatga tcttaacgag ctcacagagt ggttgtcttg 48660 gagtcttgtg tttgttctgt tgtctttgct tgctcactta ttaaacaggt ttttgtaggt 48720 agtccaagct gccctataag tcatagtagt cctgcttcag gcgcctaggt gcctggatca 48780 caggttctgt gccacgtgca tagctgctat tgattttttt tgaagttctg tggaatgaaa 48840 ccaggccatt gcccacagag tgcaaggagc tgagccgcct ctccaacgct ctgcaagttc 48900 tagcatgtgt gtgcttttga atgtcctatg gacaaaatta tgtcatctgc atatgacatt 48960 atcatgacta cgccttctca ctcaggttag aacctgttct ttcacagtgt tgaacagaaa 49020 tggtaatagg ccaggtttga ccacaagcac caaccacaaa aagaagaaaa acagtgtgtg 49080 gtggagccaa ttcctgtttt gttttctgat ctcagtgcag tatcttcaac atttaaaaac 49140 aaacgaacac taacaaaaca acaataacaa ctggtttttt gttttgtttt gtttttctct 49200 gtatagctct ggctgtcctg gaactcactt tatagaccag gctggcctca aactcacaga 49260 gagagagacc cacctgccct ggattattga ctgctgggac taaaggcatg tgccaagcac 49320 agcctgtctt caaaaacatc ttacattttt aaccatttgt ttgcatgtgc atggcccagt 49380 gcatgagagc catgccccgt gtgtggaggt ctgagggcag ctctcaggag tcagtttcca 49440 ccttccacct tcttggtttt gcttcacctc tctagtactc taggctaact accgaagagc 49500 ttctggacaa tgcttccctc taccgcccat ctccctgtgg gaccattgga ttacacgtgg 49560 gtttttccaca cgggctgggg aattgaactt gggtcgtcag gcttgctctg cgagaacttt 49620 cccctgccgt gcctatccca gggcctgtcc tccctaccta attgctaagt atgttttata 49680 tagatttgct acagacattc ttgattagat ttaggaaatt ccaaagggtt ttaaccacgt 49740 ccacatatag aaggcgttgg tgggacggac agagaaaggc aggagtgagg tgtgccagag 49800 gaattatgag ccaggctaga gctgcctctt catgttcaca gaatttctag attcttctct 49860 gccctctgct gttccagaca aaccctgatc atttgaactc cattccacct ctggccagct 49920 catcccctca ataactcgtg cacccggtgc ttcccacttc agggaggtct tgtgtgactc 49980 tcctgagcct cttcactctg attcaccctc cgtcaccaaa gcccctgtgc ttctgttcca 50040 tcctgggcgc tgcctgtcag acctctctac aagcttctca tctggggcta ccgtttcctg 50100 tctctgtaac cctgtggttc ccatcagctc ctgctctgtg aactcacaca actgggcact 50160 gccttgctct gttctgggcc tgacatcagg ctggacctgg gcatagaata cggccagcga 50220 cctttttgaa ttagataacc aggaaagcac tctcacaaat gggtcaaagt tcaaatttag 50280 tgaatttatt ttatataatt ttatcttact actttttttc ctcaaaaagt agtaggtttg 50340 ttcacagttt ggctggagct agggggaagg tgaaactttt gagctcctct gtgcctcgct 50400 tgcagcacgc tgctactgat tatggaaatg caggatattt gggtctacaa ggaacgaaaa 50460 tctgtcagaa tcttgtcctg ggaggtgact cttcctcgga atgggaaaaa aagaaaaaga 50520 aagaaagaaa ggaaggaagg aaggaaggaa ggaaggaaga aaggaagaaa aaaaggaaga 50580 aaggaagaaa ggaaggaagg aaaggacatg attggtctca ggtttcagca ccaaatgaag 50640 agagagagag agagagagag agagagagag agagagagag agagagagat gactgcttgc 50700 tcctgggtac cctggaggag atagtttatt gtagatatga gagagagcag agctagaggc 50760 agagtcaaga gtggacaaac ccagactgag atgtgccatg tgaggagagg gggaaagcaa 50820 gaaggagggg ggagagatga accaggaaca gcagccaggt ggcccagagg tacaaagaga 50880 gcagggacca aaatggtcag attatatagg gaagagcatg cccagcccct ggccagaaag 50940 cacagggtag gggtggggta tgccagctag gagggccctg taagggatag ggactgtggg 51000 atgcagggtc tgtccaggac agggactgtg ggatgcaggg agggtctgtc caggacttgg 51060 atgagttaag taagcacctc agctgcttgc ctcagtttga aacctgacat ccggctgctc 51120 tcacacccag cacagaagtc actggaaaaa ttgttgtgtt tgtttcattg agtttattgc 51180 tatctagctt ggaaatctat cctcattcag aacagaagcc tgtaaccttg aatggtgtgc 51240 cttggccctc ccacagagac aggataaata gtgaacatgt gctctggata gacaaagaga 51300 tgcatgagaa cagctcagaa ggaagcagtc ggaatgggct agggggctta gaaaagctcc 51360 ttgaaagagg ccagtggggg actggagaga ttgctcagtg gttaagagca ctgactgctc 51420 ttccggaggt cctgagttca aatcccagaa accacatggt ggctcacaac tatctgtaac 51480 aagatctgat gccctcttct agagtgtctg aagacagcta cagtgtactc acatataata 51540 aaataaataa ataaaaat aaataaatct taaaaaaaaa aaagaggtaa ctgagcccta 51600 cctgaaagaa caagtgggtg tggccatgtg gaaaggagaa gaaaagtctt cccataggga 51660 atggaaggca caaggaaaga ccccatggca ggggagggag ggtgttaagg gctggggccc 51720 agaactgaac tgggtgtaaa ccttaggttt attacttagc accatgttgc agtgggtgag 51780 ttcctaaacc tctggcctca gttccctctc tgcaaaccta aactgctgtg tgtaaagtca 51840 cctgggtgtg atgactacct tcaaagcaac aggaggttaa aacagggagg agaaacgctg 51900 gctgaatggg ggcagagtca gaagctaacg atcaccaagc ccctgctgct tggctcctaa 51960 gtggccattt ccaggactgt acttactgct cgggcctctc cgactgtggt cttggagtac 52020 ggttcagcca gggcctacct tgcctcccca gaattaatca tcactgcggg cggcgagaac 52080 gtgcccccag tgcctatcga ggaggccgtg aagatggagc tgcccatcat cagcagtgcc 52140 atgctgatcg gggaccagag gaagttcctg tccatgctgc taaccctgaa ggtgcgtcat 52200 gggctctagc tctggagctg tagtgggagg gtggtctgtg gtgtagcccg aggtacatct 52260 ggtattcaca actttatcct ggcgtcattc agccagtgcc acgaaggtgg aaattcctct 52320 gccacactgg gctcctggga tgatccttcc cacttaccag tacattccag cgtgcaaagg 52380 cctggctgcc ctttggtcac atgctcaggc cccagcatca cagccaggca gggggatgtc 52440 cctcgttgaa cagaagctaa catcatcact gttccttcct ctgcttcctg ggactcaccc 52500 accttaaaag catcactctg aaatcgagtc cacagaattc ccatcttgtt gtctgtgaca 52560 tcctcgaaga ccagagcgtg gctctgcggg ggaggggtgg accctcatcc gtcctctttt 52620 cagtgcacgc tggacccaga gacgtctgag cctacagaca gcctgaccga gcaagctgtg 52680 gagttctgcc agcgggtggg cagcaaggcc agcaccgtct ccgagatcgt ggggagagaga 52740 gacgaggctg tgtaccaggc cattcacgaa gggatccaga gggtgaacgc gaacgccgca 52800 gcccggccct accacatcca gaagtgggcc attctccaac gtgacttctc catttcaggt 52860 ggagaactgg gtaagttttt cttttcactg tccctggggc tgtatgggag ggcaaagccc 52920 atggcggtga caggcaggag tctgttagga agtagccatg ctagcagaag gtgtcagata 52980 gggttttgga cagcatctgt gctgggttca taaacctctg attctaatca acctgaaaaa 53040 taagcaggca cctccagatc cactggatgc ggccaccttg gattagaatg actaagagga 53100 aaaggtgcat ctgtcctgaa cacatgcagc tggtttcctc cccccgcccc catcctttaa 53160 gtaatagcga gtaactcctc acaaagtatt tacagtgtct ttggtttcaa acatcatgta 53220 gagatgcctt ggaagcacac aggaaagcat gtgttggtca tgtgcagata catgtcactt 53280 cacatacata gaacactggg ctttgatgct gagggaggga gctcccgaaa cctgtgatga 53340 cagcgatgag tgtgttacca gtgtgccaga gcttatcagc atggacacga agtgtttagc 53400 tctgccgtga ccatgagact ctcgtctctg ctgagttaca gtgcacacac ccatcaggta 53460 cacatcacag ccccgatgct gctgtggctg ttaaaggatc gaactgtctg gaccacagtc 53520 caagcccaga aaagcctgtt gtacagcagc ctcacccggc cagccagagg agtctcctag 53580 ctggggcatc tcgggtgctc atggcacaga acactgaaca cggctccttg gtgaggcttt 53640 caaatccact cacactgaaa ggagccacgc tatctgcagc gctgctctga tgtattggag 53700 gtactgagag tgttaatagg ttttctccac cgatgaaata agccaattca agccaaatta 53760 aaaatataaa acaggttttg gggaggagta gttcttgggc aggttcacct gtcccaagga 53820 atggagccag agaaattgcc atggagacaga cagggagagg gtgtcacaca tgcagggaga 53880 aatgcaggag gggagcatga ggggttagag agggggagga agagggaaag agagggggag 53940 aggagaaggg agaaagaaag agggagagag ggagagagag agggagaggg ggagggggag 54000 agagagagtg cgcaagagag agcaaggagc aaaatgtcag gattatgtat aatccagaag 54060 agcctctgca ggagggaaag cccagcctct gggctggaaa gttaagggta gagggcagct 54120 ggcatgccag ccatacccta taacaggtag ggactgaggg tccttgggag gccctcactt 54180 tgatatgtta aataaccaca gcagcagctt gtcctgggtc tgaaacccaa aatgtataac 54240 cagatttgtt caaattaaac acaagggcaa tcaaatgaaa gatgtcttct ctaggcagtt 54300 tataaagacc ctaaccttaa tgtctctagt gaggggctga gttatgtaca aaaaaccctt 54360 aagttaccca ggacctcaag ttgaagccct ttcatccttg tattaaagga agaaacaaaa 54420 gctaagaaaa gtgacttgag gaaaatattt tagacattat aatattgtct ggaactttaa 54480 gcatcccaac acattaaagc caagtccacc attatgaact gtccctaaaa cacgtggagc 54540 tttgtgtata agccattgct atgagacagc tactcccaag tggagaaact taaacattag 54600 acgtggagtc tagtgtggta ggacatgcct gttgtcccag cactcaagag gtgaaggcag 54660 gaagatcagg agtttaaggc tagcctctgc tacatagtga gtttggggcc agcctaggct 54720 acacaagacc ttgtcccaat taaataacaa acaagacaat aagcattatc ttcattttga 54780 tgggccaaga atcacgggca gttttctgag tggccagact gggagtttcc catgatgcta 54840 gtcagagcag accaggtgac aggcacctgt ttgcacactc atgtaattga tcactcgaac 54900 tggcttgacg ggggccagag gcttcagttc cccacaggct gcttgcacat gctagcatgg 54960 cagctggcct accctgtaag gaatgatctg agagtcctcc caagagaaac tgtgttccat 55020 tgctacaatc acttgcagct gatactctgc aaccacaggt cccacgcctg tggatctaac 55080 caacatcaga ccaaaaactt aaaaaattct tgggactgtg tagagccccc cactccccct 55140 ttctcctcag tactggaatt gactcttggg gcctccagaa caccaggcaa gagctctatc 55200 actgagcaat ctctctggtt ctctttttaa attttgagaa aagatcttac taaattcccc 55260 aggttggcct tgaactcagt gtaactcaga caagccccaa atttgtgatc ctcctgtctc 55320 agcctccgag gagctgggat tacaggccat gaagacctga ctatgcaaac tttttaatta 55380 ttactaaaca ctagtttatt tatgtagcat ttatgctgta tttgagattc tgagtagtat 55440 agagaaggct taaagaatat ggattttcat ggttttgtgc aaataccaca cagttttatg 55500 aaagactttg ggcaattttt agcttagtta tctgtggttt gaatgggttt ggcctccata 55560 gattcacgtg tttgaattta tggcccatgg ggagtggcac tattagaagg tgtggccttc 55620 tccagcacca tgtctgcctg gattctgcca tgctccccac cacgatgata atgggttaca 55680 cttctgaaac agtaagtcag ccccaattac atgttttcct ttataggagt tgccttggtc 55740 atggtgtctc tttacagcaa taaaactcta agatggtatc tgaaaaggtt tctggatgac 55800 ccatactccc tgtattctga gacagggtat gttcattcaa gggaggatca taatggggac 55860 aggtcaaggc tgtatttcat tatgtaatgg aaagaaacaa gtcacttctg cccctggaag 55920 tgacacccat catgtcctct gtaacctgac tgcagactaa gcctagttca ctgtgacaca 55980 gaggggctc tgtgggataa tgctaaaagg actcttgcct cagaagctgt acagtgtatc 56040 ctgcttctct ggcttagtac ctgacttctg gatctcacat gccatctaag tcacttttct 56100 atttctgtga taaaacacca cacccaagcc aacctataaa agaaaatgtc caatgggttc 56160 atagttccag atagttatga gtccacagta gcagagatag aaaatggaag aaatcttttg 56220 aaacttcaag cctaccctag tgctacacct cctctaaggc tgtatactct aatccttccc 56280 aaacccccc accaactggg aaaccagtta ttcaaacaca tgaccctaca ggagtcattc 56340 tcttccaaac gataccttct catgaaacca aacccaggcc acactcattt cttccaattt 56400 ttcttttagg acccaccatg aaattgaaac ggctcacggt tctggagaag tacaaagata 56460 tcatcgattc tttttatcaa gagcaaaaac agtagtcagg agccatgctt gtggcttctg 56520 ctggagacca gcccggcaag catcttcctg agcgccaggt gtctccagtg ggcaaaagaa 56580 cctgtgacaa gctgcttcca ccctttagtt cagggcagag cactgacatc tccaatgcct 56640 tcacgtgcaa gtagtttcaa ttctaaccac tgctcctgtt caggtgtcaa gccagaccat 56700 tctccttcca aaagagaagg accgctcact ttggcttcag gtccaggcag atctacaagg 56760 ttctagttca caggtctccc agggggcgct gttggagagg gaaagggatg gcaatcccat 56820 tactaatgtg tgctgcagag acagcagtca cagctcgcct gacttaattc tggcttcttg 56880 gaaactagta ccacactctg ccacctcaca cacaagaggc ttgatgctta ataaactgct 56940 gctgcctatg tgactgtatt taatatgtaa ttttcagtat taaacctttt aacagatctg 57000 caacaa 57006

Claims (13)

A kit for the diagnosis of Creutzfeldt-Jakob disease comprising an antibody or an aptamer specifically binding to an ACSBG1 (acyl-CoA synthetase bubblegum family member 1) protein.
2. The kit for the diagnosis of Creutzfeldt-Jakob disease according to claim 1, wherein the ACSBG1 comprises the amino acid sequence of SEQ ID NO: 1 or is encoded by the nucleotide sequence of SEQ ID NO: 2.
2. The kit for diagnosing Creutzfeldt-Jakob disease according to claim 1, wherein the antibody is bound to a solid substrate.
The kit for diagnosing Creutzfeldt-Jakob disease according to claim 1, further comprising a detection antibody binding to the protein.
5. The kit for diagnosing Creutzfeldt-Jakob disease according to claim 4, wherein the antibody for detection is attached to any one detection element selected from the group consisting of chromogenic enzymes, fluorescent substances, radioactive isotopes and colloids.
A biochip for the diagnosis of Creutzfeldt-Jakob disease comprising an ACSBG1 (acyl-CoA synthetase bubblegum family member 1) antibody or an aptamer.
A kit for the diagnosis of Creutzfeldt-Jakob disease comprising a primer pair that specifically amplifies an ACSBG1 (acyl-CoA synthetase bubblegum family member 1) gene, or an antisense nucleotide complementarily binding to the ACSBG1 gene.
8. The kit for diagnosing Creutzfeldt-Jakob disease according to claim 7, wherein the ACSBG1 gene comprises the nucleotide sequence of SEQ ID NO: 2.
1) measuring the expression level of ACSBG1 (acyl-CoA synthetase bubblegum family member 1) protein from the sample isolated from the subject; And
2) a step of selecting an individual having a higher expression level than the normal expression level of the protein in step 1).
[10] The method of claim 9, wherein the sample of step 1) is any one selected from the group consisting of blood, serum, plasma, cells, and tissues.
The method according to claim 9, wherein the expression level of step 1) is selected from the group consisting of Western blotting, Enzyme-linked immunosorbent assay (ELISA), RT-PCR, immunohistochemical staining, wherein the protein is detected by any one method selected from the group consisting of immunoprecipitation and immunofluorescence.
1) treating a test compound or composition with an ACSBG1 (acyl-CoA synthetase bubblegum family member 1) protein or a cell expressing the ACSBG1 protein; And
2) selecting a test compound or a composition that reduces the expression or activity of the ACSBG1 protein in the step 1); and screening for a candidate for a treatment for Creutzfeldt-Jakob disease.
13. The method according to claim 12, wherein the expression or activity of the protein in step 2) is selected from the group consisting of Western blotting, enzyme-immunochemical detection, RT-PCR, immunohistochemical staining, immunoprecipitation and immunofluorescence Wherein the method comprises screening for candidate substances for treatment of Creutzfeldt-Jakob disease.
KR1020150052375A 2015-04-14 2015-04-14 Expression validation of protein expressed uniquely by scrapie prion protein and use for development of biomarker for the diagnosis of Creutzfeldt-Jakob disease KR101802407B1 (en)

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Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014120902A1 (en) 2013-01-31 2014-08-07 Arizona Board Of Regents, A Body Corporate Of The State Of Arizona, Acting For And On Behalf Of Ari Autoantibody signature for the early detection of ovarian cancer

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014120902A1 (en) 2013-01-31 2014-08-07 Arizona Board Of Regents, A Body Corporate Of The State Of Arizona, Acting For And On Behalf Of Ari Autoantibody signature for the early detection of ovarian cancer

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Title
Ayako Moriya-Sato, et al., ‘Novel Acyl-CoA Synthetase in Adrenoleukodystrophy Target Tissues’, Biochemical and Biophysical Research Communications, (2000), 279, pp 62-68.
Yue Tang, et al., ‘Transcriptional Analysis Implicates Endoplasmic Reticulum Stress in Bovine Spongiform Encephalopathy’, PLoS ONE, (2010), Vol. 5, Issue 12, e14207.

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