KR20190030813A - A novel protein marker for diagnosing Alzheimer's disease and the use thereof - Google Patents

Abstract

According to the present invention, protein markers from blood-derived EV, which can be used for diagnosing Alzheimer′s disease, have been successfully identified. First, blood-derived EV proteins, specifically increased compared to the wild type, are identified from TG6799 mice using an unlabeled quantitative assay, and 11 new AD biomarker candidates were selected through bioinformatics and biochemical experiments. Next, the selected biomarker candidates were evaluated using samples of Alzheimer′s disease patients. Thus, the present invention provides novel protein markers for diagnosing Alzheimer′s disease and an effective method for diagnosing the same.

Description

[0001] The present invention relates to novel protein markers for the diagnosis of Alzheimer ' s disease,

The present invention relates to a biomarker for the diagnosis of Alzheimers disease (AD). More specifically, the present invention relates to a protein marker that can be used for the early diagnosis of Alzheimer's disease.

Alzheimer's disease is one of the most common diseases that cause dementia and causes cognitive impairment including thinking and memory. Drugs such as acetylcholinesterase (AChE) inhibitors or NMDA (N-methyl-D-aspartate) receptor antagonists have been used for the treatment of Alzheimer's disease, It is known to have an effect. Methods such as clinical exam and brain imaging are used to diagnose Alzheimer's disease, but it is very difficult to diagnose Alzheimer's disease at an early stage by such a diagnostic method. Studies have been continuing to diagnose Alzheimer's disease using a variety of specimens, including biofluids, and as a result, a reliable method for diagnosing Alzheimer's disease has been developed. However, these diagnostic methods are also limited in that they are only useful for diagnosis of late stage Alzheimer's disease. Although early diagnosis of the disease is essential for the effective treatment of Alzheimer's disease, there is currently no way to diagnose early stage Alzheimer's disease.

With the development of mass spectrometry techniques, subproteome research has become a next-generation paradigm for biomarker development. For example, extracellular vesicles (EVs) were considered to be merely secretomes in the early stages of their studies, but these extracellular vesicles contained cellular proteins, nucleic acids, and lipids And has been recognized as a novel signal mediator in recent years as it has been shown to exhibit properties such as a shuttling vehicle by participating in various signal mediators through paracrine or endocrine action . Although extracellular vesicles have become interesting targets for the development of biomarkers because of the inclusion of signaling proteins, nucleic acids and lipids, few studies have been conducted on the blood EVs due to their difficulty in preparing their samples.

The present inventors have endeavored to find a biomarker for diagnosis of Alzheimer's disease from the blood. As a result, they have found a novel protein biomarker that can be used for diagnosis of Alzheimer's disease in the exosome protein isolated from blood, Thereby completing the present invention.

Korean Patent Publication No. 10-2017-0005940

Choi, D. S. & Gho, Y. S. Isolation of extracellular vesicles for proteomic profiling. Methods in molecular biology 1295, 167-177, doi: 10.1007 / 978-1-4939-2550-6_14 (2015)

It is an object of the present invention to provide a protein marker for diagnosing Alzheimer's disease.

It is another object of the present invention to provide an exosome protein as a protein marker for diagnosing Alzheimer's disease.

It is another object of the present invention to provide a composition for diagnosing Alzheimer's disease.

It is another object of the present invention to provide a protein marker used for providing information necessary for diagnosis of Alzheimer's disease.

It is another object of the present invention to provide a method for screening drug candidates for treating Alzheimer's disease using protein markers.

In order to achieve the above object, the present invention provides a protein biomarker for diagnosing Alzheimer's disease isolated from blood cell extracellular follicles. In one embodiment of the present invention, we performed proteomic analysis of extracellular vesicles (EVs) in TG6799 mice and identified novel AD biomarker proteins. Specifically, in order to identify biomarker candidates existing in the blood EV, two different EV protein production methods including methanol-chloroform precipitation and non-ionic detergent-fusion gel-internal degradation were performed in parallel. Next, 18 biomarker candidate proteins specifically amplified in AD blood were screened through bioinformatics analysis. The level of increase of these proteins was then assessed by Western blotting and the clinical confidence in these was verified using blood EVs isolated from human AD patients.

The present invention also relates to a method for the diagnosis of Alzheimer's disease, comprising the steps of: a) determining the levels of HSPA8 (heat shock cognate 71 kDa protein), ITIH2 (inter-alpha trypsin inhibitor, heavy chain 2), DBP (D-binding protein), TLN1 (talin-1), CAT (catalase), STAM2 (signal transducing adapter molecule 2), PARVB (beta-parvin), GNAI2 (guanine nucleotide- At least one exosomal protein selected from the group consisting of ALAD (delta-aminolevulinic acid dehydratase), L-lactate dehydrogenase (LDA), T-complex protein 1 subunit delta (CCT4), and PRDX2 (peroxiredoxin 2) And b) if the amount of protein detected in a) is increased or decreased in the subject compared to a normal subject, the subject is judged to be an Alzheimer's disease patient.

In one embodiment of the present invention, the protein having an increased amount in the subject compared to a normal subject is a protein selected from the group consisting of HSPA8, ITIH2, GC, TLN1, CAT, STAM2, PARVB, GNAI2, ALAD, LDHA and CCT4 , The protein with a reduced amount in the subject compared to the normal person is PRDX2. In another embodiment of the present invention, protein detection detects an antigen-antibody complex formed by the protein or an immunogenic fragment thereof and an antibody thereto, and the detection may be performed by Western blotting, enzyme linked immunosorbent assay, immunoprecipitation assays, complement fixation assays, fluorescence activated cell sorters (FACS), or protein chips.

In addition, the present invention relates to a pharmaceutical composition comprising, as an active ingredient, an exosome protein or an antibody that specifically binds to an immunogenic fragment thereof, which is relatively increased or decreased in the blood of a patient suffering from Alzheimer's disease, A composition for diagnosing a disease is provided.

In one embodiment of the invention, the antibody is a monoclonal antibody, polyclonal antibody, or recombinant antibody. In another embodiment of the present invention, the composition for diagnosing Alzheimer's disease is selected from the group consisting of HSPA8 (Heat shock cognate 71 kDa protein; NCBI Genbank registration number: 15481), ITIH2 (inter-alpha trypsin inhibitor, heavy chain 2; NCBI Genbank registered (NCBI Genbank Accession Number: 14473), TLN1 (talin-1; NCBI Genbank Accession No. 21894), CAT (catalase; NCBI Genbank Accession No. 12359), STAM2 (signal (NCBI Genbank registration number: 56324), PARVB (beta-parvin; NCBI Genbank registration number: 170736), GNAI2 (guanine nucleotide-binding protein G (i) subunit alpha-2; NCBI Genbank registration number: 14678) , ALTA (delta-aminolevulinic acid dehydratase; NCBI Genbank registration number: 17025), LDHA (L-lactate dehydrogenase; NCBI Genbank registration number: 16828) and CCT4 (T-complex protein 1 subunit delta; NCBI Genbank registration number: An agent for measuring the expression level of at least one protein selected from the group consisting of It can hamhal. The agent that measures the level of expression of the protein may be an antibody specific for the protein or an immunogenic fragment thereof, wherein the antibody comprises both a monoclonal antibody, a polyclonal antibody, and a recombinant antibody. Monoclonal antibodies in the present invention may be prepared by hybridoma methods well known in the art (Kohler and Milstein (1976) European journal of Immunology 6: 511-519) or phage antibody libraries (Clarkson et al, Nature, 352: 624 Biol., 222: 58, 1-597, 1991) techniques. Polyclonal antibodies can be prepared by conventional methods, including injecting the protein antigen into an animal and obtaining serum containing the antibody from the animal. Such polyclonal antibodies can be produced from any animal, including dogs, goats, sheep, rabbits, monkeys, horses, pigs, and cows. In addition, the antibody of the present invention includes chimeric antibodies, humanized antibodies, human antibodies, and the like. The antibodies used in the present invention also include complete forms of antibodies having two full-length light chains and two full-length heavy chains and functional fragments thereof. The functional fragment of the antibody refers to a fragment having an antigen binding function and includes Fab, F (ab ') 2, F (ab') 2, Fv and the like. In addition, the present invention relates to an exosomal protein having an increased or decreased expression level in the blood of a patient suffering from Alzheimer's disease as compared with blood of a normal person, wherein the exosome protein is HSPA8, ITIH2, GC, TLN1, CAT, STAM2, PARVB, GNAI2, ALAD, LDHA, CCT4, and PRDX2. ≪ / RTI > In another aspect, the invention provides an Alzheimer's disease diagnostic kit comprising an agent that measures the level of expression of at least one protein selected from the group consisting of CCT4 and PRDX2.

In another embodiment of the present invention, the kit may comprise one or more compositions, solutions or devices suitable for analyzing protein levels, as well as agents that measure the level of the protein marker in a biological sample separated from the subject. For example, the kit may comprise a substrate, a buffer solution, a secondary antibody labeled with a detection label, and a chromogenic substrate for immunological detection of the antibody. In another embodiment of the present invention, the kit for diagnosing Alzheimer's disease provides a kit containing necessary components for performing an ELISA method such as ELISA kit, sandwich ELISA and the like. That is, the ELISA kit comprises an antibody specific for the protein marker and is capable of binding to a reagent capable of detecting the bound antibody, for example, a labeled secondary antibody, chromopores, an enzyme and its substrate or antibody And other materials that can be used.

In another embodiment of the present invention, the kit for diagnosing Alzheimer's disease may be a kit for Western blotting, immunoprecipitation assay, complement fixation assay, flow cytometry, or protein chip.

In another embodiment of the present invention, the present invention relates to a protein chip for diagnosis of Alzheimer's disease comprising the composition for diagnosing Alzheimer's disease. The protein chip is composed of one or more antibodies against the protein marker of the present invention arranged at high density on the chip. In the present invention, the detection label may be selected from the group consisting of an enzyme, a fluorescent material, a ligand, a light emitting material, a microparticle, a redox molecule, and a radioactive isotope, but is not limited thereto.

The present invention also relates to a method of preparing a blood sample, comprising the steps of: a) preparing a blood extracellular follicle from a blood sample separated from a subject; b) Expression of at least one of the genes consisting of Hspa8 , Iyih2 , Gc , Tlnl , Cat, Stam2, Parvb , Gnai2 , Alad , Ldha , Cct4 and Prdx2 , each consisting of the nucleotide sequence of SEQ ID NO: 13 to SEQ ID NO: Measuring a level; And c) determining that at least one of the genes of b) is increased or decreased in comparison to a control that is normal, it is determined that the Alzheimer ' s disease or increased risk is increased, for monitoring or diagnosis of Alzheimer's disease ≪ / RTI > In one embodiment of the present invention, the expression level of b) can be measured by any one method selected from the group consisting of RT-PCR, ELISA, Western blotting and immunohistochemistry.

The present invention also relates to a method of detecting the expression level of at least one exosome protein selected from the group consisting of HSPA8, ITIH2, GC, TLN1, CAT, STAM2, PARVB, GNAI2, ALAD, LDHA, CCT4 and PRDX2 from a biological sample isolated from a subject. ; And b) determining if the subject is at increased risk of Alzheimer ' s disease or is an Alzheimer ' s disease subject if the expression level of the protein of step a) is increased or decreased as compared to a normal control, And a method for measuring the level of exosome protein expression to provide information for disease diagnosis. Wherein the biological sample of step a) can be selected from the group consisting of bone marrow, serum, plasma and blood, and uses an antibody that specifically binds to each of the proteins for measuring the protein expression level in step a) .

Also, the present invention provides a method of treating a subject suffering from Alzheimer's disease comprising the steps of: a) b) measuring the expression level of at least one exosomal protein selected from the group consisting of HSPA8, ITIH2, GC, TLN1, CAT, STAM2, PARVB, GNAI2, ALAD, LDHA, CCT4 and PRDX2 in the extracellular follicle of step a) step; And c) selecting a test substance that increases or decreases the protein expression level of the step b) as compared to the untreated control, comprising the step of screening candidate substances for treating Alzheimer's disease. In one embodiment of the invention, the protein expression level measurement in step b) is selected from the group consisting of immunofluorescence, mass spectrometry, protein chip, Western blotting and ELISA.

In one embodiment of the invention, twelve proteins characterized by differences in expression levels from TG6799 mouse extracellular vesicles samples were screened (HSPA8, ITIH2, GC, TLN1, CAT, STAM2, PARVB, GNAI2, ALAD , LDHA, CCT4 and PRDX2), and the expression levels of HSPA8, ITIH2, GC, TLN1, CAT, STAM2, PARVB, GNAI2, ALAD, LDHA and CCT4 protein were higher in AD patients than in normal subjects 3).

As used herein, the term " diagnosis " means identifying the presence or characteristic of a disease or disease. For the purposes of the present invention, diagnosis may refer to identifying the onset of Alzheimer ' s disease, or confirming progression or intensification of the disease.

In the present invention, the term " marker for diagnosis or diagnostic marker " is a substance which distinguishes Alzheimer's disease from normal state. It is an increase or decrease in a sample of an individual having Alzheimer's disease Such as proteins, peptides, polypeptides or nucleic acids, lipids, glycolipids, or sugars (monosaccharides, disaccharides, oligosaccharides, etc.) For the purpose of the present invention, the markers for diagnosis of the present invention include HSPA8 (heat shock cognate 71 kDa protein), ITIH2 (inter-alpha trypsin inhibitor, heavy chain 2), which specifically express increased expression in a sample of Alzheimer's disease patients, , Vitamin D-binding protein (DBP), TLN1 (talin-1), CAT (catalase), STAM2 (signal transducing adapter molecule 2), PARVB (beta-parvin), GNAI2 (guanine nucleotide- alpha-2), ALAD (delta-aminolevulinic acid dehydratase), LDHA (L-lactate dehydrogenase) and CCT4 (T-complex protein 1 subunit delta), and peroxidase It refers to the protein PRDX2 (Peroxiredoxin 2). The amino acid sequence and the nucleotide sequence of the marker protein of the present invention can be confirmed in a known gene database. For example, the amino acid sequence of the human HSPA8 protein can be found in NCBI Genbank registry number 15481, which is set forth in SEQ ID NO: 1, and its base sequence is set forth in SEQ ID NO: Similarly, the amino acid sequence of the ITIH2 protein can be found in NCBI Genbank Registry # 16425, which is set forth in SEQ ID NO: 2 and its base sequence is set forth in SEQ ID NO: 14. The amino acid sequence of the DBP protein can be found at NCBI Genbank Registry No. 14473, which is set forth in SEQ ID NO: 3 and its base sequence is set forth in SEQ ID NO: The amino acid sequence of the TLN1 (talin-1) protein can be found in NCBI Genbank registration number 21894, which is shown in SEQ ID NO: 4 and the nucleotide sequence thereof is shown in SEQ ID NO: The amino acid sequence of the CAT (catalase) protein can be found in NCBI Genbank registration number 12359, which is shown in SEQ ID NO: 5 and its base sequence is shown in SEQ ID NO: The amino acid sequence of the STAM2 protein can be found in NCBI Genbank registration number 56324, which is shown in SEQ ID NO: 6, and its base sequence is shown in SEQ ID NO: 18. The amino acid sequence of the PARVB protein can be found in NCBI Genbank registry number 170736, which is set forth in SEQ ID NO: 7 and its base sequence is set forth in SEQ ID NO: 19. The amino acid sequence of the GNAI2 protein can be found in NCBI Genbank registration number 14678, which is set forth in SEQ ID NO: 8 and its base sequence is set forth in SEQ ID NO: 20. The amino acid sequence of the ALAD protein can be found in NCBI Genbank Registry No. 17025, which is shown in SEQ ID NO: 9, and the nucleotide sequence thereof is shown in SEQ ID NO: 21. The amino acid sequence of the LDHA protein can be found in NCBI Genbank Registry No. 16828, which is set forth in SEQ ID NO: 10 and its base sequence is set forth in SEQ ID NO: The amino acid sequence of the CCT4 protein can be found in NCBI Genbank registry number 12464, which is set forth in SEQ ID NO: 11 and its base sequence is set forth in SEQ ID NO: 23. The amino acid sequence of the PRDX2 protein can be found in NCBI Genbank registration number 21672, which is set forth in SEQ ID NO: 12 and its base sequence is set forth in SEQ ID NO:

The term "specimen sample" in the present invention includes samples such as tissues, cells, whole blood, plasma, serum, blood, saliva, sputum, lymphatic fluid, cerebrospinal fluid, intracellular fluid or urine in which the expression level of the protein marker of the present invention is different. But is not limited thereto. Preferably, it is an extracellular vesicle of blood. According to one embodiment of the present invention, the process of separating the protein from the blood sample can be performed using a generally known process, and the protein level can be measured by various methods of assaying the antigen-antibody complex as described above.

The present invention provides a method for measuring the expression level of at least one protein selected from the group consisting of HSPA8, ITIH2, GC, TLN1, CAT, STAM2, PARVB, GNAI2, ALAD, LDHA, CCT4, and PRDX2, And the like.

1A schematically shows a method for detecting a protein marker for diagnosis of Alzheimer's disease in blood EV of TG6799 mouse according to the present invention.
Figure 1B shows the results of confirming that monomers and tetramers of amyloid beta exist only in the brain of TG6799 mice.
FIG. 1C shows the size of an extracellular follicle using a particle analyzer. FIG.
FIG. 1D shows that a large number of protein markers of the present invention exist in the blood EV produced in the present invention.
FIG. 1E shows the presence of amyloid beta in the blood EV prepared according to the present invention.
Figures 2a and 2b show increased protein levels (71) and decreased protein levels (60) in blood EVs according to the present invention.
FIG. 3 shows that ten proteins selected in the present invention are on a line of a ball cannon representing the reliability of these protein selection criteria.
FIG. 4 is an analysis using blood EV of TG6799 mouse obtained by non-ionic detergent-fusion SDS-PAGE. As shown in FIG. 4, proteins present in the blood EV membrane were extracted and three bands specifically expressed as a result of Coomassie blue staining .
FIGS. 5A and 5B show the results of evaluation of the biomarker candidate proteins screened using TG6799 mouse blood EV, showing that these proteins were markedly increased in AD blood EV.
6A and 6B show the possibility of the AD-related protein isolated from the blood EV of TG6799 mouse using a human-derived sample as a biomarker, showing that the level of the protein significantly increased in AD blood EV.

Materials and methods

(1) Laboratory animal care

All animal experiments were carried out with the approval of the Laboratory Animal Care and Protection Committee of the Korean Brain Research Institute. The TG6799 mouse (The Jackson Laboratory) was allowed to freely consume water for 12 hours under light / dark cycle conditions.

(2) blood Extracellular follicle  Produce

EDTA (ethylenediaminetetraacetic acid) and centrifugation to prepare plasma, and the plasma was diluted 10-fold with HBSS buffer (cat # 14175095, Invitrogen). Next, after incubating at 4 DEG C for 60 minutes, the diluted solution was centrifuged at 12,000 rpm at 4 DEG C for 20 minutes using a table top centrifuge. The resulting pellet was dissolved in 1 ml of HBS and centrifuged at 120,000 x g for 90 minutes at 4 占 폚. The precipitated pellet was resuspended in 200 [mu] l of HBS. The exosomal protein concentrate was measured with a bicinchoninic acid assay (BCA) assay (Cat # 23225, ThermoFisher scientific).

(3) Blood for mass analysis Extracellular follicle Peptides  Produce

A methanol-chloroform process was performed. Briefly, 100 [mu] l of extracellular vesicle was mixed with 400 [mu] l of methanol and vortexed thoroughly. The mixture was centrifuged at 9,000 xg for 5 minutes. Next, 100 μl of chloroform was added and vortexed vigorously. After centrifugation at 9,000 xg for 5 minutes, 300 L of chloroform was added and vortexed vigorously. The mixture was centrifuged at 16,000 xg for 5 minutes and then the upper layer was carefully removed. 300 [mu] l of methanol was added to the upper layer, followed by gently tapping and mixing. After centrifugation at 16,000 x g for 5 minutes, the supernatant was removed. Then, after drying for 5 minutes, the pellets were collected. For in-solution digestion, the pellet was resuspended in 100 μl of 6 M urea in 40 mM NH ₄ HCO ₃ . After incubation for 1 hour at room temperature with 5 mM dithiothreitol (DTT), iodoacetamide (IAA) was treated at room temperature for 30 minutes in a dark room. Trypsin (Promega, Madison, WI, USA) at a ratio of 1: 20 to protein was treated at 37 占 폚 for 16 hours. After drying using a freeze dryer, a desalting column (# 89873, Thermo Fisher Scientific, San Jose, CA, USA) was used according to the manufacturer's instructions. To perform the nonionic detergent fusion SDS-PAGE method, blood EVs were resuspended in 1% DDM (n-dodecyl-ss-D-maltoside) and incubated at 37 DEG C for 10 minutes. 5% Beta-mercaptoethanol and SDS (Sodium Dodecyl Sulfate) sample buffer were mixed with the resuspension mixture and boiled for 10 minutes, and the sample was analyzed by SDS-PAGE. The gel plugs were washed with 30% ACN (acetonitrile) and treated with 10 mM DTT at 56 ° C for 30 minutes. After washing the gel plug with 100% ACN, 100 mM IAA was treated in the dark room at room temperature for 30 minutes. After the gel plug was washed with 100% ACN, the gel plug was dried with a freeze dryer, and treated with trypsin (1:50 ratio) at 37 ° C for 16 hours.

(4) Mass analysis and DB analysis

Trypsin degradation products were analyzed by Q-exited plus hybrid quadrupole orbit-trap mass spectrometer (Thermo Fisher Scientific, San Jose, CA, USA) with a nano-electrospray ion source. Chromatographic separation of the peptides was carried out using Acclaim PepMap ^™ 100 (75 μm × 2 cm, Thermo Fisher Scientific, San Jose, CA, USA) and Acclaim PepMap ^™ RSLC (75 μm × 15 cm, 100 resin, Thermo Fisher scientific, (San Jose, Calif., USA) equipped with an Ultimate 3000 RSLCnano system (Thermo Fisher Scientific, San Jose, CA, USA). Peptides were loaded from RS auto-sampler and separated by a linear gradient of ACN / water at a flow rate of 300 nl / min containing 0.1% formic acid. The LC eluent was electron sprayed from the analytical column and a voltage of 2.0 kV was applied through the liquid interface of the nano spray source. Peptide mixtures were separated using ACN with gradient from 5% to 40% for 40 minutes. The assay consisted of a full MS scan in the range of 350 to 2000 m / z and a data-dependent MS / MS (MS2) on the ten strongest ions from the whole MS scan. The mass analyzer is programmed to capture in a data dependent mode. The mass spectrometer calibration was performed with the proposed calibration solution according to the manufacturer's instructions. In the present invention, proteomic analysis was performed by using three replicates of biological samples per condition, and performing two replicates for each sample (technical replicates). To perform the database search, the tandem mass spectra were processed with the Thermo Fisher Scientific Proteome Discoverer software version 2.1.1.21. The spectrum data was searched for a mouse Uniprot database (release version 2016_06). The analysis flow used here consists of four nodes: Spectrum Files (data input), Spectrum Selector (spectrum and feature retrieval), Sequest HT (sequence database search), and Percolator (PSM Validation and FDR analysis ). All identified proteins had a false discovery rate (FDR) of ≤ 1%, calculated at the peptide level. The evaluation was based on q-value. The search parameters allowed tryptic specificity of up to 2 missed cleavage counts with methylthio-modification of cysteine as methionine oxidation and fixed modification as a dynamic modification . The mass search parameters for +1, +2, and +3 ions included mass error tolerances of 20 ppm for precursor ions and 0.6 Da for fragment ions.

(5) Western blotting

Blood EVs were mixed with SDS sample buffer (Bio-Rad) and 5% beta-mercaptoethanol and boiled for 10 minutes. Brain tissue was removed from the mice, washed with ice-cold PBS and homogenized with a radioimmunoprecipitation assay buffer (Thermo fisher) containing 1x Halt protease and protease inhibitor cocktail. The resulting homogenate was sonicated and cell debris was removed by centrifugation at 13,000 x g for 15 minutes at 4 ° C. Protein concentration was measured with a BCA assay (ThermoFisher scientific) and the lysate was mixed with SDS sample buffer. Proteins were separated by SDS-PAGE, transferred to PVDF (polyvinylidene fluoride) membrane using Bio-Rad wet transfer system, blocked with PBS-T containing 5% skim milk for 30 minutes, Lt; 0 > C for 16 hours. After washing the membrane three times with PBS-T, the blot was incubated with HRP (horseradish peroxidase) -conjugated anti-mouse or anti-rabbit antibody for 1 hour at room temperature. Next, the membrane was washed with PBS-T (phosphate buffered saline with Tween 20, Thermo Fisher Scientific) and developed with an ECL Western blotting system (GE Healthcare). Inter-alpha trypsin inhibitor heavy chain 2 (ab118257, Abcam), vitamin D-binding protein (ab153922, Abcam), catalase (ab52477, Abcam), talin-1 (4021S, Cell Signaling Technology), heat shock cognate 71 kDa protein (ab154415, Abcam), signal transducing adapter molecule (SH3 domain and ITAM motif) 2 (226050, United States Biological), Peroxiredoxin 2 (ab71533, Abcam), Beta-parvin (sc-375581, Santa Cruz Biotechnology) binding protein G (i) subunit alpha-2 (sc-13534, Santa Cruz Biotechnology), porphobilinogen synthase (sc-271585, Santa Cruz Biotechnology), L-lactate dehydrogenase A chain (PA5-23036, Thermo Fisher Scientific) Western blot analysis for the complex protein 1 subunit delta (ab129072, Abcam), CD9 (# 553758, BD Biosciences), CD63 (sc-15363, Santa Cruz Biotechnology), CD81 (ab33697, Abcam), and Alix Blotting was performed.

(6) Statistical analysis

The relative amounts of the proteins identified in the control and AD groups were compared against PSMs (peptide spectrum matches) from each condition. For each protein, the p-value of the quantitative difference between the two conditions was evaluated by a Chi-square test. The chi-square test value was measured with a G test (test of goodness of fit; see Equation 1 below) with the set freedom as follows:

F2 * log (f ^ 2) - f1 * log (f ^ 1) - f2 * log (f ^

Where f1 and f2 represent PSMs from the LC-MS / MS results of blood EVs in the control and AD groups, respectively, and f ^ 1 and f ^ 2 represent predicted PSMs. The predicted PSMs normalized to the ratio of the sum of PSMs. The predicted PSMs were evaluated as the sum of the PSMs in both conditions, and the sum of the PSMs of each condition was multiplied by the ratio of the total PSMs. For exclusively identified proteins, 0.25 PSM values were given for the non-detected conditions for the evaluation of the p-value.

Example

Example  1: Blood from TG6799 mouse Extracellular follicle  Produce

The process for producing extracellular blood vesicles from TG6799 mice is schematically illustrated in Figure 1A. To confirm amyloid beta expression observed only in TG6799 mice, we performed Western blotting using a whole brain lysate of TG6799 mouse (6 months old, available from The Jackson Laboratory). We have found that monomers and tetramers of amyloid beta are present only in the TG6799 brain (FIG. 1b). In order to find AD-dependent proteins in blood EV, we first isolated blood EV from wild-type and TG6799 mouse blood. Plasma of blood was prepared, and then the plasma was subjected to concentration gradient ultracentrifugation to obtain blood EV. For the evaluation of the quality of the blood EV, the vesicle size was analyzed using a particle analyzer (Nanosight, manufacturer). As a result, most of the 100 ㎚ sized follicles were found (Fig. 1C). Next, the present inventors confirmed the follicular biomarker protein from the blood EV prepared above. Solid pellet biomarker proteins such as CD9, CD63, CD81 and Alix were present in very large amounts in the blood EVs prepared above (Fig. 1d). Since it has been reported that amyloid beta is present in several vesicles, the present inventors confirmed the presence or absence of amyloid beta in the prepared blood EV. As a result, it was confirmed that amyloid beta was present in the prepared blood EV (Fig. 1E). Thus, the present inventors confirmed that high quality blood EVs was isolated from TG6799 mouse and wild type mouse.

Example  2: Proteome analysis of blood EV using methanol-chloroform precipitation

As a first method of producing an EV proteome, the obtained EV was treated with methanol-chloroform to precipitate the EV protein. Next, the resulting precipitate was subjected to in-solution digestion to prepare a trypsin-degrading peptide. The trypsin degrading peptides from the blood EVs were analyzed by LC-electrospray (ESI) -MS / MS. Sequest search engine-applied Proteome discoverer 2.1 searches the Uniprot database to identify 317 proteins from wild-type and TG6799 mice. 71 enhanced proteins and 60 reduced proteins were selected as non-labeled quantitative analysis, satisfying the selection criteria of log ₂ > 3 fold change or high statistical significance (P <0.05) And Figure 2b). From the proteomic results, we finally selected 10 proteins as AD biomarker candidates (Table 1).

Uniprot Gene ID Protein name % Coverage
(aver) Fold change
(log2) G3X977 Itih2 Inter-alpha trypsin inhibitor, heavy chain 2 8.6 5.91 P21614 Gc Vitamin D-binding protein 10.4 5.00 H7BWY6 Rbp4 Retinol-binding protein 4 9.0 4.58 P24270 Cat Catalase 5.4 4.58 B2RPV6 Mmrn1 Multimerin-1 2.1 4.32 Q99JI6 Rap1b Ras-related protein Rap-1b 14.7 4.00 Q80TM2 Tln1 Talin-1 1.2 4.00 P63017 Hspa8 Heat shock cognate 71 kDa protein 2.0 3.58 Q3TGH8 Stam2 Signal transducing adapter molecule 2 3.6 3.00 Q3UNK5 Tgfb1 Transforming growth factor beta-1 5.5 3.00

The 10 selected proteins are selected from the group consisting of inter-alpha trypsin inhibitor, heavy chain 2 (Itih2), vitamin D-binding protein (DBP), retinol-binding protein 4 (Rbp4), catalase (Rap1b), tallin-1 (Tln1), Hspa8 (Heat shock cognate 71 kDa protein), and ras-related protein ras- Stam2 (signal transducing adapter molecule 2) and Tgfb1 (transforming growth factor beta-1). A volcano plot of the 10 selected proteins showed that these proteins are on a line of the ball cano type indicating the reliability of the selection criteria (Fig. 3).

Example  3: Proteome analysis of blood EV using nonionic detergent-fusion SDS-PAGE

There are several reports that non-ionic detergents are suitable for the extraction of membrane proteins. The present inventors extracted proteins present in the EV membrane using non-ionic detergent DDM (n-dodecyl D-maltoside). Blood EV prepared by concentration gradient ultracentrifugation was added to 1% DDM and hydrophobic proteins including membrane proteins were extracted. Next, the extract was analyzed by SDS-PAGE. We found three band regions of TG6799 mouse blood EV specifically present in Coomassie Brilliant Blue staining (Figure 4). For the three band regions, trypsin substrate gel-internal degradation was performed. A total of 160 proteins were identified by LC-MS / MS analysis using trypsin digested peptides (results not shown). From these total proteins, the following 8 proteins were selected as biomarker candidates with high confidence in proteome parameters: β-parvin (PARVB), delta-aminolevulinic acid dehydratase (ALAD), guanine nucleotide-binding protein G (i) L-lactate dehydrogenase (LDH), malate dehydrogenase (MDH2), T-complex protein 1 subunit beta (CCT2) and T-complex protein 1 subunit delta (CCT4) (Table 2).

Uniprot Gene ID Description Coverage # PSMs # Unique
peptides Q3UGT9 Parvb Beta-parvin 9.9 3 3 P10518 Alad Delta-aminolevulinic acid dehydratase (porphobilinogen synthase) 9.1 3 3 P08752 Gnai2 Guanine nucleotide-binding protein G (i) subunit alpha-2 7.3 2 2 Q8CG19 Ltbp1 Latent-transforming growth factor beta-binding protein 1 2.6 4 3 A0A1B0GSX0 Ldha L-lactate dehydrogenase 15.2 5 5 P08249 Mdh2 Malate dehydrogenase, mitochondrial 15.7 4 4 Q542X7 Cct2 T-complex protein 1 subunit beta 7.7 2 2 P80315 Cct4 T-complex protein 1 subunit delta 6.1 3 3

Example  4: Mouse Blood from EV Screened Biomarker  Evaluation of candidate proteins

Biochemical experiments were conducted to evaluate the potential as an AD biomarker. After the blood EV was prepared in the same manner as in the proteome analysis, Western blotting was performed on the biomarker candidate. Six proteins including heat shock cognate 71 kDa protein, inter-alpha trypsin inhibitor, heavy chain 2, vitamin D-binding protein, talin-1, catalase and signal transducing adapter molecule 2 were found to be significantly increased in AD blood EV (Fig. 5A). Peroxiredoxin 2 (PRDX2), identified as reduced in proteome analysis, was significantly reduced in TG6799 blood EVs (Fig. 5A). Western blotting was also performed on the biomarker candidates identified in the nonionic detergent fusion SDS-PAGE method. Five proteins including β-parvin, guanine nucleotide-binding protein G (i) subunit alpha-2, delta-aminolevulinic acid dehydratase, L-lactate dehydrogenase and T-complex protein 1 subunit delta were significantly increased in AD blood EV (Fig. 5B). These Western blotting results suggest that the eleven proteins are highly likely as AD biomarkers.

Example  5: Using the blood EV of AD patients who were separated in stages Biomarker  Evaluation of candidate proteins

A sample isolated from humans was used to evaluate the possibility as an AD biomarker. First, three types of blood samples were prepared: normal, early AD, and late AD. All groups were over 60 years old. After the blood EV was prepared in the same manner as in the proteome analysis, western blotting was performed on each biomarker. L-lactate dehydrogenase A chain, ras-related protein Rap-1b, retinol-binding protein 4, porphobilinogen synthase, guanine nucleotide-binding protein G (i) The level of six proteins including the subunit alpha-2 was significantly increased in AD blood EV (Figs. 6A and 6B). These results suggest that these six proteins are highly likely to be biomarkers that can be directly applied to AD diagnosis.

The present invention separates protein markers which can be used for diagnosis of Alzheimer's disease from blood EV, and can be used for diagnosis of Alzheimer's disease. In particular, the protein markers of the present invention provide the possibility of diagnosing early stage Alzheimer's disease. Therefore, the present invention can be used clinically very useful for diagnosis and treatment of Alzheimer's disease.

<110> Daegu Gyeongbuk Institute of Science and Technology <120> A novel protein marker for diagnosing Alzheimer's disease and the          use thereof <130> IKR0117P03KR <160> 24 <170> KoPatentin 3.0 <210> 1 <211> 646 <212> PRT <213> Artificial Sequence <220> <223> Heat shock cognate 71 kDa protein <400> 1 Met Ser Lys Gly Pro Ala Val Gly Ile Asp Leu Gly Thr Thr Tyr Ser   1 5 10 15 Cys Val Gly Val Phe Gln His Gly Lys Val Glu Ile Ile Ala Asn Asp              20 25 30 Gln Gly Asn Arg Thr Thr Pro Ser Tyr Val Ala Phe Thr Asp Thr Glu          35 40 45 Arg Leu Ile Gly Asp Ala Ala Lys Asn Gln Val Ala Met Asn Pro Thr      50 55 60 Asn Thr Val Phe Asp Ala Lys Arg Leu Ile Gly Arg Arg Phe Asp Asp  65 70 75 80 Ala Val Val Gln Ser Asp Met Lys His Trp Pro Phe Met Val Val Asn                  85 90 95 Asp Ala Gly Arg Pro Lys Val Gln Val Glu Tyr Lys Gly Glu Thr Lys             100 105 110 Ser Phe Tyr Pro Glu Glu Val Ser Ser Met Val Leu Thr Lys Met Lys         115 120 125 Glu Ile Ala Glu Ala Tyr Leu Gly Lys Thr Val Thr Asn Ala Val Val     130 135 140 Thr Val Pro Ala Tyr Phe Asn Asp Ser Gln Arg Gln Ala Thr Lys Asp 145 150 155 160 Ala Gly Thr Ile Ala Gly Leu Asn Val Leu Arg Ile Ile Asn Glu Pro                 165 170 175 Thr Ala Ala Ile Ala Tyr Gly Leu Asp Lys Lys Val Gly Ala Glu             180 185 190 Arg Asn Val Leu Ile Phe Asp Leu Gly Gly Gly Thr Phe Asp Val Ser         195 200 205 Ile Leu Thr Ile Glu Asp Gly Ile Phe Glu Val Lys Ser Thr Ala Gly     210 215 220 Asp Thr His Leu Gly Gly Glu Asp Phe Asp Asn Arg Met Val Asn His 225 230 235 240 Phe Ile Ala Glu Phe Lys Arg Lys His Lys Lys Asp Ile Ser Glu Asn                 245 250 255 Lys Arg Ala Val Arg Arg Leu Arg Thr Ala Cys Glu Arg Ala Lys Arg             260 265 270 Thr Leu Ser Ser Thr Gln Ala Ser Ile Glu Ile Asp Ser Leu Tyr         275 280 285 Glu Gly Ile Asp Phe Tyr Thr Ser Ile Thr Arg Ala Arg Phe Glu Glu     290 295 300 Leu Asn Ala Asp Leu Phe Arg Gly Thr Leu Asp Pro Val Glu Lys Ala 305 310 315 320 Leu Arg Asp Ala Lys Leu Asp Lys Ser Gln Ile His Asp Ile Val Leu                 325 330 335 Val Gly Gly Ser Thr Arg Ile Pro Lys Ile Gln Lys Leu Leu Gln Asp             340 345 350 Phe Phe Asn Gly Lys Glu Leu Asn Lys Ser Ile Asn Pro Asp Glu Ala         355 360 365 Val Ala Tyr Gly Ala Ala Val Gln Ala Ile Leu Ser Gly Asp Lys     370 375 380 Ser Glu Asn Val Gln Asp Leu Leu Leu Leu Asp Val Thr Pro Leu Ser 385 390 395 400 Leu Gly Ile Gly Thr Ala Gly Gly Val Met Thr Val Leu Ile Lys Arg                 405 410 415 Asn Thr Thr Ile Pro Thr Lys Gln Thr Gln Thr Phe Thr Thr Tyr Ser             420 425 430 Asp Asn Gln Pro Gly Val Leu Ile Gln Val Tyr Glu Gly Glu Arg Ala         435 440 445 Met Thr Lys Asp Asn Asn Leu Leu Gly Lys Phe Glu Leu Thr Gly Ile     450 455 460 Pro Pro Ala Pro Arg Gly Val Pro Gln Ile Glu Val Thr Phe Asp Ile 465 470 475 480 Asp Ala Asn Gly Ile Leu Asn Val Ser Ala Val Asp Lys Ser Thr Gly                 485 490 495 Lys Glu Asn Lys Ile Thr Ile Thr Asn Asp Lys Gly Arg Leu Ser Lys             500 505 510 Glu Asp Ile Glu Arg Met Met Gln Glu Ala Glu Lys Tyr Lys Ala Glu         515 520 525 Asp Glu Lys Gln Arg Asp Lys Val Ser Ser Lys Asn Ser Leu Glu Ser     530 535 540 Tyr Ala Phe Asn Met Lys Ala Thr Val Glu Asp Glu Lys Leu Gln Gly 545 550 555 560 Lys Ile Asn Asp Glu Asp Lys Gln Lys Ile Leu Asp Lys Cys Asn Glu                 565 570 575 Ile Ile Ser Trp Leu Asp Lys Asn Gln Thr Ala Glu Lys Glu Glu Phe             580 585 590 Glu His Gln Gln Lys Glu Leu Glu Lys Val Cys Asn Pro Ile Ile Thr         595 600 605 Lys Leu Tyr Gln Ser Ala Gly Gly Met Pro Gly Gly Met Pro Gly Gly     610 615 620 Phe Pro Gly Gly Gly Ala Pro Pro Ser Gly Gly Ala Ser Ser Gly Pro 625 630 635 640 Thr Ile Glu Glu Val Asp                 645 <210> 2 <211> 950 <212> PRT <213> Artificial Sequence <220> <223> Inter-alpha trypsin inhibitor, heavy chain 2 <400> 2 Met Ser Ser Lys Met Gln Arg Pro Val Cys Leu Leu Ile Trp Leu Phe   1 5 10 15 Leu Leu Glu Ala Gln Ala Phe Glu Ile Pro Ile Asn Gly Asn Ser Glu              20 25 30 Phe Ala Glu Tyr Ser Asp Leu Val Glu Leu Ala Pro Asp Lys Leu Pro          35 40 45 Phe Val Gln Glu Asn Gly Arg His Gln Arg Ser Leu Pro Glu Glu Ser      50 55 60 Gly Glu Glu Thr Asp Thr Val Asp Pro Val Thr Leu Tyr Ser Tyr Lys  65 70 75 80 Val Gln Ser Thr Ile Thr Ser Arg Val Ala Thr Thr Thr Ile Gln Ser                  85 90 95 Lys Leu Val Asn Asn Ser Pro Leu Pro Gln Ser Val Val Phe Asp Val             100 105 110 Gln Ile Pro Lys Gly Ala Phe Ile Ser Asn Phe Thr Met Thr Val Asn         115 120 125 Gly Met Thr Phe Thr Ser Ser Ile Lys Glu Lys Thr Val Gly Arg Ala     130 135 140 Leu Tyr Ser Gln Ala Arg Ala Lys Gly Lys Thr Ala Gly Trp Val Arg 145 150 155 160 Ser Arg Thr Leu Asp Met Glu Asn Phe Asn Thr Glu Val Asn Ile Pro                 165 170 175 Pro Gly Ala Lys Val Gln Phe Glu Leu His Tyr Gln Glu Val Lys Trp             180 185 190 Arg Lys Leu Gly Ser Tyr Glu His Lys Ile His Leu Gln Pro Gly Lys         195 200 205 Leu Ala Lys His Leu Glu Val Asn Val Trp Ile Ile Glu Pro Gln Gly     210 215 220 Met Arg Phe Leu His Val Pro Asp Thr Phe Glu Gly His Phe Gln Gly 225 230 235 240 Val Pro Val Ile Ser Lys Gly Gln Gln Lys Ala His Val Ser Phe Lys                 245 250 255 Pro Thr Val Ala Gln Gln Arg Lys Cys Pro Asn Cys Thr Glu Thr Ala             260 265 270 Val Asn Gly Glu Leu Val Val Met Tyr Asp Val Asn Arg Glu Glu Lys         275 280 285 Ala Gly Glu Leu Glu Val Phe Asn Gly Tyr Phe Val His Phe Phe Ala     290 295 300 Pro Glu Asn Leu Asp Pro Ile Pro Lys Asn Ile Leu Phe Val Ile Asp 305 310 315 320 Val Ser Gly Ser Met Trp Gly Ile Lys Met Lys Gln Thr Val Glu Ala                 325 330 335 Met Lys Thr Ile Leu Asp Asp Leu Arg Thr Asp Asp Gln Phe Ser Val             340 345 350 Val Asp Phe Asn His Asn Val Arg Thr Trp Arg Asn Asp Leu Val Ser         355 360 365 Ala Thr Lys Thr Gln Ile Ala Asp Ala Lys Arg Tyr Ile Glu Lys Ile     370 375 380 Gln Pro Ser Gly Gly Thr Asn Ile Asn Glu Ala Leu Leu Arg Ala Ile 385 390 395 400 Phe Ile Leu Asn Glu Ala Ser Asn Met Gly Leu Leu Asn Pro Asp Ser                 405 410 415 Val Ser Leu Ile Ile Leu Val Ser Asp Gly Asp Pro Thr Val Gly Glu             420 425 430 Leu Lys Leu Ser Lys Ile Gln Lys Asn Val Lys Gln Ser Ile Gln Asp         435 440 445 Asn Ile Ser Leu Phe Ser Leu Gly Ile Gly Phe Asp Val Asp Tyr Asp     450 455 460 Phe Leu Lys Arg Leu Ser Asn Glu Asn Arg Gly Ile Ala Gln Arg Ile 465 470 475 480 Tyr Gly Asn Gln Asp Thr Ser Ser Gln Leu Lys Lys Phe Tyr Asn Gln                 485 490 495 Val Ser Thr Pro Leu Leu Arg Asn Val Gln Phe Asn Tyr Pro Gln Ala             500 505 510 Ser Val Thr Asp Val Thr Gln Asn Asn Phe His Asn Tyr Phe Gly Gly         515 520 525 Ser Glu Ile Val Val Ala Gly Lys Phe Asp Pro Ser Lys Leu Thr Glu     530 535 540 Val Gln Ser Ile Ile Thr Ala Thr Ser Ala Asn Thr Glu Leu Val Leu 545 550 555 560 Glu Thr Leu Ser Gln Met Asp Asp Leu Glu Glu Phe Leu Ser Lys Asp                 565 570 575 Lys His Ala Asp Pro Asp Phe Thr Lys Lys Leu Trp Ala Tyr Leu Thr             580 585 590 Ile Asn Gln Leu Leu Ala Glu Arg Ser Leu Ala Pro Thr Ala Ala Ile         595 600 605 Lys Arg Lys Ile Thr Lys Thr Ile Leu Gln Met Ser Leu Asp His His     610 615 620 Ile Val Thr Pro Leu Thr Ala Met Val Ile Glu Asn Asp Ala Gly Asp 625 630 635 640 Glu Arg Met Leu Ala Asp Ser Pro Pro Gln Asp His Ser Cys Cys Ser                 645 650 655 Gly Ala Leu Tyr Tyr Gly Thr Lys Val Ala Ser Gly Pro Ile Pro Ser             660 665 670 Trp Ala Asn Pro Ser Pro Thr Pro Met Ser Ala Met Leu Ala Val Gly         675 680 685 Ala Lys Pro Leu Glu Ser Thr Pro Pro Thr His Leu Asn Gln Val Glu     690 695 700 Asn Asp Pro His Phe Ile Ile Tyr Leu Pro Lys Ser Lys Arg Asn Ile 705 710 715 720 Cys Phe Asn Ile Asp Ser Glu Pro Gly Lys Ile Leu Ser Leu Val Ser                 725 730 735 Asp Pro Glu Ser Gly Ile Val Val Asn Gly Gln Leu Ile Gly Ala Lys             740 745 750 Arg Ala Glu Asn Gly Lys Leu Ser Thr Tyr Phe Gly Lys Leu Gly Phe         755 760 765 Tyr Phe Gln Lys Glu Gly Met Lys Ile Glu Ile Ser Thr Glu Thr Ile     770 775 780 Thr Leu Ser Ser Gly Ser Ser Thr Ser Arg Leu Ser Trp Ser Asp Thr 785 790 795 800 Ala His Leu Gly Asn Ser Arg Val Leu Ile Ser Val Lys Lys Glu Lys                 805 810 815 Ser Val Thr Leu Thr Leu Asn Lys Glu Leu Phe Ser Val Leu Leu             820 825 830 His Arg Val Trp Arg Lys His Pro Val Asn Val Asp Phe Leu Gly Ile         835 840 845 Tyr Ala Pro Pro Ile Asp Lys Phe Ser Pro Arg Val His Gly Leu Leu     850 855 860 Gly Gln Phe Met Gln Glu Pro Ala Ile His Ile Phe Asn Glu Arg Pro 865 870 875 880 Gly Lys Glu Pro Gly Lys Pro Glu Ala Ser Met Glu Val Lys Gly His                 885 890 895 Lys Leu Thr Val Thr Arg Gly Leu Gln Lys Asp Tyr Arg Thr Asp Ile             900 905 910 Val Phe Gly Thr Asp Val Pro Cys Trp Phe Val His Asn Ser Gly Lys         915 920 925 Gly Phe Ile Asp Gly His Tyr Lys Asp Tyr Phe Val Pro Gln Leu Tyr     930 935 940 Ser Phe Leu Lys Arg Pro 945 950 <210> 3 <211> 476 <212> PRT <213> Artificial Sequence <220> <223> Vitamin D-binding protein <400> 3 Met Lys Arg Val Leu Leu Leu Leu Ala Leu Ala Phe Gly His Ala   1 5 10 15 Leu Glu Arg Gly Arg Asp Tyr Glu Lys Asp Lys Val Cys Asn Glu Leu              20 25 30 Ala Met Leu Gly Lys Glu Asp Phe Arg Ser Leu Ser Leu Ile Leu Tyr          35 40 45 Ser Arg Lys Phe Ser Ser Ser Thr Phe Glu Gln Val Asn Gln Leu Val      50 55 60 Lys Glu Val Val Ser Leu Thr Glu Glu Cys Cys Ala Glu Gly Ala Asp  65 70 75 80 Pro Thr Cys Tyr Asp Thr Arg Thr Ser Glu Leu Ser Val Lys Ser Cys                  85 90 95 Glu Ser Asp Ala Pro Phe Pro Val His Pro Gly Thr Pro Glu Cys Cys             100 105 110 Thr Lys Glu Gly Leu Glu Arg Lys Leu Cys Met Ala Ala Leu Ser His         115 120 125 Gln Pro Gln Glu Phe Pro Thr Tyr Val Glu Pro Thr Asn Asp Glu Ile     130 135 140 Cys Glu Ala Phe Arg Arg Asp Pro Lys Gly Phe Ala Asp Gln Phe Leu 145 150 155 160 Tyr Glu Tyr Ser Ser Asn Tyr Gly Gln Ala Pro Leu Pro Leu Leu Val                 165 170 175 Ala Tyr Thr Lys Asn Tyr Leu Ser Met Val Gly Ser Cys Cys Thr Ser             180 185 190 Ala Asn Pro Thr Val Cys Phe Val Lys Glu Arg Leu Gln Met Lys His         195 200 205 Leu Ser Leu Leu Thr Thr Met Ser Asn Arg Val Cys Ser Gln Tyr Ala     210 215 220 Ala Tyr Gly Lys Glu Lys Ser Arg Leu Ser His Leu Ile Lys Leu Ala 225 230 235 240 Gln Lys Val Pro Thr Ala Asn Leu Glu Asn Val Leu Pro Leu Ala Glu                 245 250 255 Asp Phe Thr Glu Ile Leu Ser Arg Cys Cys Glu Ser Thr Ser Glu Asp             260 265 270 Cys Met Ala Ser Glu Leu Pro Glu His Thr Ile Lys Ile Cys Gln Asn         275 280 285 Leu Ser Lys Lys Asn Ser Lys Phe Glu Glu Cys Cys Gln Glu Asn Thr     290 295 300 Pro Met Asn Ile Phe Met Cys Thr Tyr Phe Met Pro Ala Ala Glu Pro 305 310 315 320 Leu Gln Leu Pro Ala Ile Lys Leu Pro Thr Gly Lys Asp Leu Cys Gly                 325 330 335 Gln Ser Thr Gln Ala Met Asp Gln Tyr Thr Phe Glu Leu Ser Arg             340 345 350 Arg Thr Gln Val Pro Glu Val Phe Leu Ser Lys Val Leu Glu Pro Thr         355 360 365 Leu Lys Thr Leu Arg Glu Cys Cys Asp Thr Gln Asp Ser Val Ala Cys     370 375 380 Phe Ser Thr Gln Ser Pro Leu Leu Lys Arg Gln Leu Thr Ser Phe Ile 385 390 395 400 Glu Lys Gly Gln Glu Met Cys Ala Asp Tyr Ser Glu Asn Thr Phe Thr                 405 410 415 Glu Tyr Lys Lys Lys Leu Ala Glu Arg Leu Arg Thr Lys Thr Pro Asn             420 425 430 Thr Ser Pro Ala Glu Leu Lys Asp Met Val Glu Lys His Ser Asp Phe         435 440 445 Ala Ser Lys Cys Cys Ser Ile Asn Ser Pro Pro Leu Tyr Cys Ser Ser     450 455 460 Gln Ile Asp Ala Glu Met Ile Asp Thr Leu Gln Ser 465 470 475 <210> 4 <211> 2541 <212> PRT <213> Artificial Sequence <220> <223> Talin-1 <400> 4 Met Val Ala Leu Ser Leu Lys Ile Ser Ile Gly Asn Val Val Lys Thr   1 5 10 15 Met Gln Phe Glu Pro Ser Thr Met Val Tyr Asp Ala Cys Arg Met Ile              20 25 30 Arg Glu Arg Ile Pro Glu Ala Leu Ala Gly Pro Pro Asn Asp Phe Gly          35 40 45 Leu Phe Leu Ser Asp Asp Asp Pro Lys Lys Gly Ile Trp Leu Glu Ala      50 55 60 Gly Lys Ala Leu Asp Tyr Tyr Met Leu Arg Asn Gly Asp Thr Met Glu  65 70 75 80 Tyr Arg Lys Lys Gln Arg Pro Leu Lys Ile Arg Met Leu Asp Gly Thr                  85 90 95 Val Lys Thr Ile Met Val Asp Asp Ser Lys Thr Val Thr Asp Met Leu             100 105 110 Met Thr Ile Cys Ala Arg Ile Gly Ile Thr Asn His Asp Glu Tyr Ser         115 120 125 Leu Val Arg Glu Leu Met Glu Glu Lys Lys Asp Glu Gly Thr Gly Thr     130 135 140 Leu Arg Lys Asp Lys Thr Leu Leu Arg Asp Glu Lys Lys Met Glu Lys 145 150 155 160 Leu Lys Gln Lys Leu His Thr Asp Asp Glu Leu Asn Trp Leu Asp His                 165 170 175 Gly Arg Thr Leu Arg Glu Gln Gly Val Glu Glu His Glu Thr Leu Leu             180 185 190 Leu Arg Arg Lys Phe Phe Tyr Ser Asp Gln Asn Val Asp Ser Arg Asp         195 200 205 Pro Val Gln Leu Asn Leu Leu Tyr Val Gln Ala Arg Asp Asp Ile Leu     210 215 220 Asn Gly Ser His Pro Val Ser Phe Asp Lys Ala Cys Glu Phe Ala Gly 225 230 235 240 Phe Gln Cys Gln Ile Gln Phe Gly Pro His Asn Glu Gln Lys His Lys                 245 250 255 Ala Gly Phe Leu Asp Leu Lys Asp Phe Leu Pro Lys Glu Tyr Val Lys             260 265 270 Gln Lys Gly Glu Arg Lys Ile Phe Gln Ala His Lys Asn Cys Gly Gln         275 280 285 Met Ser Glu Ile Glu Ala Lys Val Arg Tyr Val Lys Leu Ala Arg Ser     290 295 300 Leu Lys Thr Tyr Gly Val Ser Phe Phe Leu Val Lys Glu Lys Met Lys 305 310 315 320 Gly Lys Asn Lys Leu Val Pro Arg Leu Leu Gly Ile Thr Lys Glu Cys                 325 330 335 Val Met Arg Val Asp Glu Lys Thr Lys Glu Val Ile Gln Glu Trp Ser             340 345 350 Leu Thr Asn Ile Lys Arg Trp Ala Ala Ser Pro Lys Ser Phe Thr Leu         355 360 365 Asp Phe Gly Asp Tyr Gln Asp Gly Tyr Tyr Ser Val Gln Thr Thr Glu     370 375 380 Gly Glu Gln Ile Ala Gln Leu Ile Ala Gly Tyr Ile Asp Ile Ile Leu 385 390 395 400 Lys Lys Lys Lys Ser Lys Asp His Phe Gly Leu Glu Gly Asp Glu Glu                 405 410 415 Ser Thr Met Leu Glu Asp Ser Val Ser Pro Lys Lys Ser Thr Val Leu             420 425 430 Gln Gln Gln Tyr Asn Arg Val Gly Lys Val Glu His Gly Ser Val Ala         435 440 445 Leu Pro Ala Ile Met Arg Ser Gly Ala Ser Gly Pro Glu Asn Phe Gln     450 455 460 Val Gly Ser Met Pro Pro Ala Gln Gln Gln Ile Thr Ser Gly Gln Met 465 470 475 480 His Arg Gly His Met Pro Pro Leu Thr Ser Ala Gln Gln Ala Leu Thr                 485 490 495 Gly Thr Ile Asn Ser Ser Met Gln Ala Val Gln Ala Ala Gln Ala Thr             500 505 510 Leu Asp Asp Phe Glu Thr Leu Pro Pro Leu Gly Gln Asp Ala Ala Ser         515 520 525 Lys Ala Trp Arg Lys Asn Lys Met Asp Glu Ser Lys His Glu Ile His     530 535 540 Ser Gln Val Asp Ala Ile Thr Ala Gly Thr Ala Ser Val Val Asn Leu 545 550 555 560 Thr Ala Gly Asp Pro Ala Glu Thr Asp Tyr Thr Ala Val Gly Cys Ala                 565 570 575 Val Thr Thr Ile Ser Ser Asn Leu Thr Glu Met Ser Arg Gly Val Lys             580 585 590 Leu Leu Ala Leu Leu Glu Asp Glu Gly Gly Asn Gly Arg Pro Leu         595 600 605 Leu Gln Ala Ala Lys Gly Leu Ala Gly Ala Val Ser Glu Leu Leu Arg     610 615 620 Ser Ala Gln Pro Ala Ser Ala Glu Pro Arg Gln Asn Leu Leu Gln Ala 625 630 635 640 Ala Gly Asn Val Gly Gln Ala Ser Gly Glu Leu Leu Gln Gln Ile Gly                 645 650 655 Glu Ser Asp Thr Asp Pro His Phe Gln Asp Val Leu Met Gln Leu Ala             660 665 670 Lys Ala Val Ala Ser Ala Ala Ala Leu Val Leu Lys Ala Lys Ser         675 680 685 Val Ala Gln Arg Thr Glu Asp Ser Gly Leu Gln Thr Gln Val Ile Ala     690 695 700 Ala Ala Thr Gln Cys Ala Leu Ser Thr Ser Gln Leu Val Ala Cys Thr 705 710 715 720 Lys Val Val Ala Pro Thr Ile Ser Ser Pro Val Cys Gln Glu Gln Leu                 725 730 735 Val Glu Ala Gly Arg Leu Val Ala Lys Ala Val Glu Gly Cys Val Ser             740 745 750 Ala Ser Gln Ala Ala Thr Glu Asp Gly Gln Leu Leu Arg Gly Val Gly         755 760 765 Ala Ala Ala Thr Ala Val Thr Gln Ala Leu Asn Glu Leu Leu Gln His     770 775 780 Val Lys Ala His Ala Thr Gly Ala Gly Pro Ala Gly Arg Tyr Asp Gln 785 790 795 800 Ala Thr Asp Thr Ile Leu Thr Val Thr Glu Asn Ile Phe Ser Ser Met                 805 810 815 Gly Asp Ala Gly Glu Met Val Arg Gln Ala Arg Ile Leu Ala Gln Ala             820 825 830 Thr Ser Asp Leu Val Asn Ala Ile Lys Ala Asp Ala Glu Gly Glu Ser         835 840 845 Asp Leu Glu Asn Ser Arg Lys Leu Leu Ser Ala Ala Lys Ile Leu Ala     850 855 860 Asp Ala Thr Ala Lys Met Val Glu Ala Ala Lys Gly Ala Ala Ala His 865 870 875 880 Pro Asp Ser Glu Glu Gln Gln Gln Arg Leu Arg Glu Ala Ala Glu Gly                 885 890 895 Leu Arg Met Ala Thr Asn Ala Ala Ala Gln Asn Ala Ile Lys Lys Lys             900 905 910 Leu Val Gln Arg Leu Glu His Ala Ala Lys Gln Ala Ala Ala Ser Ala         915 920 925 Thr Gln Thr Ile Ala Ala Ala Gln His Ala Ala Ser Ala Pro Lys Ala     930 935 940 Ser Ala Gly Pro Gln Pro Leu Leu Val Gln Ser Cys Lys Ala Val Ala 945 950 955 960 Glu Gln Ile Pro Leu Leu Val Gln Gly Val Arg Gly Ser Gln Ala Gln                 965 970 975 Pro Asp Ser Pro Ser Ala Gln Leu Ala Leu Ile Ala Ala Ser Gln Ser             980 985 990 Phe Leu Gln Pro Gly Gly Lys Met Val Ala Ala Ala Lys Ala Ser Val         995 1000 1005 Pro Thr Ile Gln Asp Gln Ala Ser Ala Met Gln Leu Ser Gln Cys Ala    1010 1015 1020 Lys Asn Leu Gly Thr Ala Leu Ala Glu Leu Arg Thr Ala Ala Gln Lys 1025 1030 1035 1040 Ala Gln Glu Ala Cys Gly Pro Leu Glu Met Asp Ser Ala Leu Ser Val                1045 1050 1055 Val Gln Asn Leu Glu Lys Asp Leu Gln Glu Ile Lys Ala Ala Ala Arg            1060 1065 1070 Asp Gly Lys Leu Lys Pro Leu Pro Gly Glu Thr Met Glu Lys Cys Thr        1075 1080 1085 Gln Asp Leu Gly Asn Ser Thr Lys Ala Val Ser Ser Ala Ile Ala Lys    1090 1095 1100 Leu Leu Gly Glu Ile Ala Gly Gly Asn Glu Asn Tyr Ala Gly Ile Ala 1105 1110 1115 1120 Ala Arg Asp Val Ala Gly Gly Leu Arg Ser Leu Ala Gln Ala Ala Arg                1125 1130 1135 Gly Val Ala Leu Thr Ser Asp Pro            1140 1145 1150 Asp Thr Ala Ser Asp Val Leu Asp Lys Ala Ser Ser Leu Ile Glu Glu        1155 1160 1165 Ala Lys Lys Ala Ser Gly His Pro Gly Asp Pro Glu Ser Gln Gln Arg    1170 1175 1180 Leu Ala Gln Val Ala Lys Ala Val Thr Gln Ala Leu Asn Arg Cys Val 1185 1190 1195 1200 Ser Cys Leu Pro Gly Gln Arg Asp Val Asp Asn Ala Leu Arg Ala Val                1205 1210 1215 Gly Asp Ala Ser Lys Arg Leu Leu Ser Asp Ser Leu Pro Pro Ser Thr            1220 1225 1230 Gly Thr Phe Gln Glu Ala Gln Ser Arg Leu Asn Glu Ala Ala Ala Gly        1235 1240 1245 Leu Asn Gln Ala Ala Thr Glu Leu Val Gln Ala Ser Arg Gly Thr Pro    1250 1255 1260 Gln Asp Leu Ala Arg Ala Ser Gly Arg Phe Gly Gln Asp Phe Ser Thr 1265 1270 1275 1280 Phe Leu Glu Ala Gly Val Glu Met Ala Gly Gln Ala Pro Ser Gln Glu                1285 1290 1295 Asp Arg Ala Gln Val Val Ser Asn Leu Lys Gly Ile Ser Met Ser Ser            1300 1305 1310 Ser Lys Leu Leu Leu Ala Ala Lys Ala Leu Ser Thr Asp Pro Ala Ser        1315 1320 1325 Pro Asn Leu Lys Ser Gln Leu Ala Ala Ala Ala Arg Ala Val Thr Asp    1330 1335 1340 Ser Ile Asn Gln Leu Ile Thr Met Cys Thr Gln Gln Ala Pro Gly Gln 1345 1350 1355 1360 Lys Glu Cys Asp Asn Ala Leu Arg Gln Leu Glu Thr Val Arg Glu Leu                1365 1370 1375 Leu Glu Asn Pro Val Gln Pro Ile Asn Asp Met Ser Tyr Phe Gly Cys            1380 1385 1390 Leu Asp Ser Val Met Glu Asn Ser Lys Val Leu Gly Glu Ala Met Thr        1395 1400 1405 Gly Ile Ser Gln Asn Ala Lys Asn Gly Asn Leu Pro Glu Phe Gly Asp    1410 1415 1420 Ala Ile Ala Thr Ala Ser Lys Ala Leu Cys Gly Phe Thr Glu Ala Ala 1425 1430 1435 1440 Ala Gln Ala Ala Tyr Leu Val Gly Val Ser Asp Pro Asn Ser Gln Ala                1445 1450 1455 Gly Gln Gln Gly Leu Val Glu Pro Thr Gln Phe Ala Arg Ala Asn Gln            1460 1465 1470 Ala Ile Gln Met Ala Cys Gln Ser Leu Gly Glu Pro Gly Cys Thr Gln        1475 1480 1485 Ala Gln Val Leu Ser Ala Ala Thr Ile Val Ala Lys His Thr Ser Ala    1490 1495 1500 Leu Cys Asn Ser Cys Arg Leu Ala Ser Ala Arg Thr Ala Asn Pro Thr 1505 1510 1515 1520 Ala Lys Arg Gln Phe Val Gln Ser Ala Lys Glu Val Ala Asn Ser Thr                1525 1530 1535 Ala Asn Leu Val Lys Thr Ile Lys Ala Leu Asp Gly Asp Phe Thr Glu            1540 1545 1550 Glu Asn Arg Ala Gln Cys Arg Ala Ala Thr Ala Pro Leu Leu Glu Ala        1555 1560 1565 Val Asp Asn Leu Ser Ala Phe Ala Ser Asn Pro Glu Phe Ser Ser Val    1570 1575 1580 Pro Ala Gln Ile Ser Pro Glu Gly Arg Ala Ala Met Glu Pro Ile Val 1585 1590 1595 1600 Ile Ser Ala Lys Thr Met Leu Glu Ser Ala Gly Gly Leu Ile Gln Thr                1605 1610 1615 Ala Arg Ala Leu Ala Val Asn Pro Arg Asp Pro Pro Arg Trp Ser Val            1620 1625 1630 Leu Ala Gly His Ser Arg Thr Val Ser Asp Ser Ile Lys Lys Leu Ile        1635 1640 1645 Thr Ser Met Arg Asp Lys Ala Pro Gly Gln Leu Glu Cys Glu Thr Ala    1650 1655 1660 Ile Ala Leu Asn Ser Cys Leu Arg Asp Leu Asp Gln Ala Ser Leu 1665 1670 1675 1680 Ala Ala Val Ser Gln Gln Leu Ala Pro Arg Glu Gly Ile Ser Gln Glu                1685 1690 1695 Ala Leu His Thr Gln Met Leu Thr Ala Val Gln Glu Ile Ser His Leu            1700 1705 1710 Ile Glu Pro Leu Ala Ser Ala Ala Arg Ala Glu Ala Ser Gln Leu Gly        1715 1720 1725 His Lys Val Ser Gln Met Ala Gln Tyr Phe Glu Pro Leu Thr Leu Ala    1730 1735 1740 Ala Val Gly Ala Ala Ser Lys Thr Leu Ser His Pro Gln Gln Met Ala 1745 1750 1755 1760 Leu Leu Asp Gln Thr Lys Thr Leu Ala Glu Ser Ala Leu Gln Leu Leu                1765 1770 1775 Tyr Thr Ala Lys Glu Ala Gly Gly Asn Pro Lys Gln Ala Ala His Thr            1780 1785 1790 Gln Glu Ala Leu Glu Glu Ala Val Gln Met Met Thr Glu Ala Val Glu        1795 1800 1805 Asp Leu Thr Thr Thr Leu Asn Glu Ala Ala Ser Ala Ala Gly Val Val    1810 1815 1820 Gly Gly Met Val Asp Ser Ile Thr Gln Ala Ile Asn Gln Leu Asp Glu 1825 1830 1835 1840 Gly Pro Met Gly Asp Pro Glu Gly Ser Phe Val Asp Tyr Gln Thr Thr                1845 1850 1855 Met Val Arg Thr Ala Lys Ala Ile Ala Val Thr Val Gln Glu Met Val            1860 1865 1870 Thr Lys Ser Asn Thr Ser Pro Glu Glu Leu Gly Pro Leu Ala Asn Gln        1875 1880 1885 Leu Thr Ser Asp Tyr Gly Arg Leu Ala Ser Gln Ala Lys Pro Ala Ala    1890 1895 1900 Val Ala Ala Glu Asn Glu Glu Ile Gly Ala His Ile Lys His Arg Val 1905 1910 1915 1920 Gln Glu Leu Gly His Gly Cys Ser Ala Leu Val Thr Lys Ala Gly Ala                1925 1930 1935 Leu Gln Cys Ser Pro Ser Asp Val Tyr Thr Lys Lys Glu Leu Ile Glu            1940 1945 1950 Cys Ala Arg Arg Val Ser Glu Lys Val Ser His Val Val Leu Ala Ala Leu        1955 1960 1965 Gln Ala Gly Asn Arg Gly Thr Gln Ala Cys Ile Thr Ala Ala Ser Ala    1970 1975 1980 Val Ser Gly Ile Ile Ala Asp Leu Asp Thr Thr Ile Met Phe Ala Thr 1985 1990 1995 2000 Ala Gly Thr Leu Asn Arg Glu Gly Ala Glu Thr Phe Ala Asp His Arg                2005 2010 2015 Glu Gly Ile Leu Lys Thr Ala Lys Val Leu Val Glu Asp Thr Lys Val            2020 2025 2030 Leu Val Gln Asn Ala Ala Gly Ser Gln Glu Lys Leu Ala Gln Ala Ala        2035 2040 2045 Gln Ser Ser Val Ala Thr Ile Thr Arg Leu Ala Asp Val Val Lys Leu    2050 2055 2060 Gly Ala Ala Ser Leu Gly Ala Glu Asp Pro Glu Thr Gln Val Val Leu 2065 2070 2075 2080 Ile Asn Ala Val Lys Asp Val Ala Lys Ala Leu Gly Asp Leu Ile Ser                2085 2090 2095 Ala Thr Lys Ala Ala Aly Gly Lys Val Gly Asp Asp Ala Val Trp            2100 2105 2110 Gln Leu Lys Asn Ser Ala Lys Val Met Val Thr Asn Val Thr Ser Leu        2115 2120 2125 Leu Lys Thr Val Lys Ala Val Glu Asp Glu Ala Thr Lys Gly Thr Arg    2130 2135 2140 Ala Leu Glu Ala Thr Thr Glu His Ile Arg Gln Glu Leu Ala Val Phe 2145 2150 2155 2160 Cys Ser Pro Glu Pro Pro Ala Lys Thr Ser Thr Pro Glu Asp Phe Ile                2165 2170 2175 Arg Met Thr Lys Gly Ile Thr Met Ala Thr Ala Lys Ala Val Ala Ala            2180 2185 2190 Gly Asn Ser Cys Arg Gln Glu Asp Val Ile Ala Thr Ala Asn Leu Ser        2195 2200 2205 Arg Arg Ala Ile Ala Asp Met Leu Arg Ala Cys Lys Glu Ala Ala Phe    2210 2215 2220 His Pro Glu Val Ala Pro Asp Val Arg Leu Arg Ala Leu His Tyr Gly 2225 2230 2235 2240 Arg Glu Cys Ala Asn Gly Tyr Leu Glu Leu Leu Asp His Val Leu Leu                2245 2250 2255 Thr Leu Gln Lys Pro Asn Pro Asp Leu Lys Gln Gln Leu Thr Gly His            2260 2265 2270 Ser Lys Arg Val Ala Gly Ser Val Thr Glu Leu Ile Gln Ala Ala Glu        2275 2280 2285 Ala Met Lys Gly Thr Glu Trp Val Asp Pro Glu Asp Pro Thr Val Ile    2290 2295 2300 Glu Aslan Glu Aslan Glu Aslan Glu Aslan Gul Ala Aslan 2305 2310 2315 2320 Lys Lys Leu Glu Gln Leu Lys Pro Arg Ala Lys Pro Lys Glu Ala Asp                2325 2330 2335 Glu Ser Leu Asn Phe Glu Glu Gln Ile Leu Glu Ala Ala Lys Ser Ile            2340 2345 2350 Ala Ala Ala Thr Ser Ala Leu Val Lys Ala Ala Ala Ser Ala Ala Gln Arg        2355 2360 2365 Glu Leu Val Ala Gln Gly Lys Val Gly Ala Ile Pro Ala Asn Ala Leu    2370 2375 2380 Asp Asp Gly Gln Trp Ser Gln Gly Leu Ile Ser Ala Ala Arg Met Val 2385 2390 2395 2400 Ala Ala Ala Thr Asn Ala Ala Ala Val Gln                2405 2410 2415 Gly His Ala Ser Gln Glu Lys Leu Ile Ser Ser Ala Lys Gln Val Ala            2420 2425 2430 Ala Ser Thr Ala Gln Leu Leu Val Ala Cys Lys Val Lys Ala Asp Gln        2435 2440 2445 Asp Ser Glu Ala Met Lys Arg Leu Gln Ala Ala Gly Asn Ala Val Lys    2450 2455 2460 Arg Ala Ser Asp Asn Leu Val Lys Ala Ala Gln Lys Ala Ala Ala Phe 2465 2470 2475 2480 Glu Asp Gln Glu Asn Glu Thr Val Val Val Lys Glu Lys Met Val Gly                2485 2490 2495 Gly Ile Ala Gln Ile Ale Ala Gln Glu Glu Met Leu Arg Lys Glu            2500 2505 2510 Arg Glu Leu Glu Glu Ala Arg Lys Lys Leu Ala Gln Ile Arg Gln Gln        2515 2520 2525 Gln Tyr Lys Phe Leu Pro Ser Glu Leu Arg Asp Glu His    2530 2535 2540 <210> 5 <211> 527 <212> PRT <213> Artificial Sequence <220> <223> Catalase <400> 5 Met Ser Asp Ser Arg Asp Pro Ala Ser Asp Gln Met Lys Gln Trp Lys   1 5 10 15 Glu Gln Arg Ala Ser Gln Arg Pro Asp Val Leu Thr Thr Gly Gly Gly              20 25 30 Asn Pro Ile Gly Asp Lys Leu Asn Ile Met Thr Ala Gly Ser Arg Gly          35 40 45 Pro Leu Leu Val Gln Asp Val Val Phe Thr Asp Glu Met Ala His Phe      50 55 60 Asp Arg Glu Arg Ile Pro Glu Arg Val Val His Ala Lys Gly Ala Gly  65 70 75 80 Ala Phe Gly Tyr Phe Glu Val Thr His Asp Ile Thr Arg Tyr Ser Lys                  85 90 95 Ala Lys Val Phe Glu His Ile Gly Lys Arg Thr Pro Ile Ala Val Arg             100 105 110 Phe Ser Thr Val Thr Gly Glu Ser Gly Ser Ala Asp Thr Val Arg Asp         115 120 125 Pro Arg Gly Phe Ala Val Lys Phe Tyr Thr Glu Asp Gly Asn Trp Asp     130 135 140 Leu Val Gly Asn Asn Thr Pro Ile Phe Phe Ile Arg Asp Ala Ile Leu 145 150 155 160 Phe Pro Ser Phe Ile His Ser Gln Lys Arg Asn Pro Gln Thr His Leu                 165 170 175 Lys Asp Pro Asp Met Val Trp Asp Phe Trp Ser Leu Arg Pro Glu Ser             180 185 190 Leu His Gln Val Ser Phe Leu Phe Ser Asp Arg Gly Ile Pro Asp Gly         195 200 205 His Arg His Met Asn Gly Tyr Gly Ser His Thr Phe Lys Leu Val Asn     210 215 220 Ala Asp Gly Glu Ala Val Tyr Cys Lys Phe His Tyr Lys Thr Asp Gln 225 230 235 240 Gly Ile Lys Asn Leu Pro Val Gly Glu Ala Gly Arg Leu Ala Gln Glu                 245 250 255 Asp Pro Asp Tyr Gly Leu Arg Asp Leu Phe Asn Ala Ile Ala Asn Gly             260 265 270 Asn Tyr Pro Ser Trp Thr Phe Tyr Ile Gln Val Met Thr Phe Lys Glu         275 280 285 Ala Glu Thr Phe Pro Phe Asn Pro Phe Asp Leu Thr Lys Val Trp Pro     290 295 300 His Lys Asp Tyr Pro Leu Ile Pro Val Gly Lys Leu Val Leu Asn Lys 305 310 315 320 Asn Pro Val Asn Tyr Phe Ala Glu Val Glu Gln Met Ala Phe Asp Pro                 325 330 335 Ser Asn Met Pro Pro Gly Ile Glu Pro Ser Pro Asp Lys Met Leu Gln             340 345 350 Gly Arg Leu Phe Ala Tyr Pro Asp Thr His Arg Arg His Arg Leu Gly Pro         355 360 365 Asn Tyr Leu Gln Ile Pro Val Asn Cys Pro Tyr Arg Ala Arg Val Ala     370 375 380 Asn Tyr Gln Arg Asp Gly Pro Met Cys Met His Asp Asn Gln Gly Gly 385 390 395 400 Ala Pro Asn Tyr Tyr Pro Asn Ser Phe Ser Ala Pro Glu Gln Gln Arg                 405 410 415 Ser Ala Leu Glu His Ser Val Gln Cys Ala Val Asp Val Lys Arg Phe             420 425 430 Asn Ser Ala Asn Glu Asp Asn Val Thr Gln Val Arg Thr Phe Tyr Thr         435 440 445 Lys Val Leu Asn Glu Glu Glu Arg Lys Arg Leu Cys Glu Asn Ile Ala     450 455 460 Gly His Leu Lys Asp Ala Gln Leu Phe Ile Gln Lys Lys Ala Val Lys 465 470 475 480 Asn Phe Thr Asp Val His Pro Asp Tyr Gly Ala Arg Ile Gln Ala Leu                 485 490 495 Leu Asp Lys Tyr Asn Ala Glu Lys Pro Lys Asn Ala Ile His Thr Tyr             500 505 510 Thr Gln Ala Gly Ser His Met Ala Ala Lys Gly Lys Ala Asn Leu         515 520 525 <210> 6 <211> 523 <212> PRT <213> Artificial Sequence <220> <223> Signal transducing adapter molecule 2 <400> 6 Met Pro Leu Phe Thr Ala Asn Pro Phe Glu Gln Asp Val Glu Lys Ala   1 5 10 15 Thr Asn Glu Tyr Asn Thr Thr Glu Asp Trp Ser Leu Ile Met Asp Ile              20 25 30 Cys Asp Arg Val Gly Ser Thr Pro Ser Gly Ala Lys Asp Cys Leu Lys          35 40 45 Ala Ile Met Lys Arg Val Asn His Lys Val Pro His Val Ala Leu Gln      50 55 60 Ala Leu Thr Leu Leu Gly Ala Cys Val Ala Asn Cys Gly Lys Ile Phe  65 70 75 80 His Leu Glu Val Cys Ser Arg Asp Phe Ala Thr Glu Val Arg Ser Val                  85 90 95 Ile Lys Asn Lys Ala His Pro Lys Val Cys Glu Lys Leu Lys Ser Leu             100 105 110 Met Val Glu Trp Ser Glu Glu Phe Gln Lys Asp Pro Gln Phe Ser Leu         115 120 125 Ile Ser Ala Thr Ile Lys Ser Met Lys Glu Glu Gly Val Thr Phe Pro     130 135 140 Ser Ala Gly Ser Gln Thr Val Ala Ala Ala Lys Asn Gly Thr Ser 145 150 155 160 Leu Asn Lys Asn Lys Glu Asp Glu Asp Ile Ala Lys Ala Ile Glu Leu                 165 170 175 Ser Leu Gln Glu Gln Lys Gln Gln Tyr Thr Glu Thr Lys Ala Leu Tyr             180 185 190 Pro Pro Ala Glu Ser Gln Leu Asn Asn Lys Ala Ala Arg Arg Val Arg         195 200 205 Ala Leu Tyr Asp Phe Glu Ala Val Glu Asp Asn Glu Leu Thr Phe Lys     210 215 220 His Gly Glu Leu Ile Thr Val Leu Asp Asp Ser Asp Ala Asn Trp Trp 225 230 235 240 Gln Gly Glu Asn His Arg Gly Thr Gly Leu Phe Pro Ser Asn Phe Val                 245 250 255 Thr Asp Leu Ser Thr Glu Val Glu Thr Ala Thr Val Asp Lys Leu             260 265 270 Asn Val Ile Asp Asp Asp Val Glu Glu Ile Lys Lys Ser Glu Pro Glu         275 280 285 Pro Val Tyr Ile Asp Glu Gly Lys Met Asp Arg Ala Leu Gln Ile Leu     290 295 300 Gln Ser Ile Asp Pro Lys Glu Ser Lys Pro Asp Ser Gln Asp Leu Leu 305 310 315 320 Asp Leu Glu Asp Val Cys Gln Gln Met Gly Pro Met Ile Asp Glu Lys                 325 330 335 Leu Glu Glu Ile Asp Arg Lys His Ser Glu Leu Ser Glu Leu Asn Val             340 345 350 Lys Val Leu Glu Ala Leu Asp Leu Tyr Asn Lys Leu Val Asn Glu Ala         355 360 365 Pro Val Tyr Ser Val Tyr Ser Lys Leu His Pro Ala His Tyr Pro Pro     370 375 380 Ala Ala Ala Gly Val Val Gln Thr Tyr Pro Val Gln Ser His Gly 385 390 395 400 Gly Asn Tyr Leu Gly His Gly Ile His Gln Val Ser Val Ala Gln Asn                 405 410 415 Tyr Asn Leu Gly Pro Asp Pro Met Gly Ser Leu Arg Ser Leu Pro Pro             420 425 430 Asn Met Asn Ser Val Thr Ala His Thr Val Gln Pro Pro Tyr Leu Ser         435 440 445 Thr Gly Gln Asp Thr Val Ser Asn Pro Ser Tyr Met Asn Gln Ser Ser     450 455 460 Arg Leu Gln Ala Ala Gly Thr Ala Ala Tyr Thr Gln Pro Val Gly 465 470 475 480 Met Ser Thr Asp Val Ser Ser Phe Gln Asn Thr Ala Ser Gly Leu Pro                 485 490 495 Gln Leu Ala Gly Phe Pro Ala Gln             500 505 510 Pro Gln Ala Ser Tyr His Gln Gln Pro Leu Leu         515 520 <210> 7 <211> 365 <212> PRT <213> Artificial Sequence <220> <223> Beta-parvin <400> 7 Met Ser Ser Ala Pro Pro Arg Ser Pro Thr Pro Arg Ala Pro Lys Met   1 5 10 15 Lys Lys Asp Glu Ser Phe Leu Gly Lys Leu Gly Gly Thr Leu Ala Arg              20 25 30 Lys Lys Lys Thr Arg Glu Val Thr Asp Leu Gln Glu Glu Gly Lys Ser          35 40 45 Ala Ile Asn Ser Pro Ala Pro Ala Leu Val Asp Ile His Pro Glu      50 55 60 Asp Thr Gln Leu Glu Glu Asn Glu Glu Arg Thr Met Ile Asp Pro Thr  65 70 75 80 Ser Arg Glu Asp Pro Lys Phe Lys Glu Leu Val Lys Val Leu Leu Asp                  85 90 95 Trp Ile Asn Asp Val Leu Ala Glu Glu Arg Ile Ile Val Lys Gln Leu             100 105 110 Glu Glu Asp Leu Tyr Asp Gly Gln Val Leu Gln Lys Leu Leu Glu Lys         115 120 125 Leu Ala His Cys Lys Leu Asn Val Ala Glu Val Thr Gln Ser Glu Ile     130 135 140 Gly Gln Lys Gln Lys Leu Gln Thr Val Leu Glu Ala Val Gln Asp Leu 145 150 155 160 Leu Arg Pro His Gly Trp Pro Leu Arg Trp Asn Val Asp Ser Ile His                 165 170 175 Gly Lys Asn Leu Val Ala Ile Leu His Leu Leu Val Ser Leu Ala Met             180 185 190 His Phe Arg Ala Pro Ile His Leu Pro Glu His Val Thr Val Gln Val         195 200 205 Val Val Val Arg Lys Arg Glu Gly Leu Leu His Ser Ser His Ile Ser     210 215 220 Glu Glu Leu Thr Thr Thr Thr Glu Ile Met Met Gly Arg Phe Glu Arg 225 230 235 240 Asp Ala Phe Asp Thr Leu Phe Asp His Ala Pro Asp Lys Leu Asn Leu                 245 250 255 Val Lys Lys Ser Leu Ile Thr Phe Val Asn Lys His Leu Asn Lys Leu             260 265 270 Asn Leu Glu Val Thr Asp Leu Glu Thr Gln Phe Ala Asp Gly Val Tyr         275 280 285 Leu Val Leu Leu Leu Gly Leu Leu Glu Asp Tyr Phe Val Pro Leu His     290 295 300 Asn Phe Tyr Leu Thr Pro Asp Ser Phe Asp Gln Lys Val His Asn Val 305 310 315 320 Ala Phe Ala Phe Glu Leu Met Leu Asp Gly Gly Leu Lys Lys Pro Lys                 325 330 335 Ala Arg Pro Glu Asp Val Val Asn Leu Asp Leu Lys Ser Thr Leu Arg             340 345 350 Val Leu Tyr Thr Leu Phe Thr Lys Tyr Lys Asp Val Glu         355 360 365 <210> 8 <211> 355 <212> PRT <213> Artificial Sequence <220> <223> Guanine nucleotide-binding protein G (i) subunit alpha-2 <400> 8 Met Gly Cys Thr Val Ser Ala Glu Asp Lys Ala Ala Ala Glu Arg Ser   1 5 10 15 Lys Met Ile Asp Lys Asn Leu Arg Glu Asp Gly Glu Lys Ala Ala Arg              20 25 30 Glu Val Lys Leu Leu Leu Leu Gly Ala Gly Glu Ser Gly Lys Ser Thr          35 40 45 Ile Val Lys Gln Met Lys Ile Ile His Glu Asp Gly Tyr Ser Glu Glu      50 55 60 Glu Cys Arg Gln Tyr Arg Ala Val Val Tyr Ser Asn Thr Ile Gln Ser  65 70 75 80 Ile Met Ala Ile Val Lys Ala Met Gly Asn Leu Gln Ile Asp Phe Ala                  85 90 95 Asp Pro Gln Arg Ala Asp Asp Ala Arg Gln Leu Phe Ala Leu Ser Cys             100 105 110 Ala Ala Glu Glu Glu Gly Met Leu Pro Glu Asp Leu Ser Gly Val Ile         115 120 125 Arg Arg Leu Trp Ala Asp His Gly Val Gln Ala Cys Phe Gly Arg Ser     130 135 140 Arg Glu Tyr Gln Leu Asn Asp Ser Ala Ala Tyr Tyr Leu Asn Asp Leu 145 150 155 160 Glu Arg Ile Ala Gln Ser Asp Tyr Ile Pro Thr Gln Gln Asp Val Leu                 165 170 175 Arg Thr Arg Val Lys Thr Thr Gly Ile Val Glu Thr His Phe Thr Phe             180 185 190 Lys Asp Leu His Phe Lys Met Phe Asp Val Gly Gly Gln Arg Ser Glu         195 200 205 Arg Lys Lys Trp Ile His Cys Phe Glu Gly Val Thr Ala Ile Ile Phe     210 215 220 Cys Val Ala Leu Ser Ala Tyr Asp Leu Val Leu Ala Glu Asp Glu Glu 225 230 235 240 Met Asn Arg Met His Glu Ser Met Lys Leu Phe Asp Ser Ile Cys Asn                 245 250 255 Asn Lys Trp Phe Thr Asp Thr Ser Ile Leu Phe Leu Asn Lys Lys             260 265 270 Asp Leu Phe Glu Glu Lys Ile Thr Gln Ser Ser Leu Thr Ile Cys Phe         275 280 285 Pro Glu Tyr Thr Gly Ala Asn Lys Tyr Asp Glu Ala Ala Ser Tyr Ile     290 295 300 Gln Ser Lys Phe Glu Asp Leu Asn Lys Arg Lys Asp Thr Lys Glu Ile 305 310 315 320 Tyr Thr His Phe Thr Cys Ala Thr Asp Thr Lys Asn Val Gln Phe Val                 325 330 335 Phe Asp Ala Val Thr Asp Val Ile Ile Lys Asn Asn Leu Lys Asp Cys             340 345 350 Gly Leu Phe         355 <210> 9 <211> 330 <212> PRT <213> Artificial Sequence <220> <223> Delta-aminolevulinic acid dehydratase (porphobilinogen synthase) <400> 9 Met His His Gln Ser Val Leu His Ser Gly Tyr Phe His Pro Leu Leu   1 5 10 15 Arg Ser Trp Gln Thr Ala Ala Ser Thr Val Ser Ala Ser Asn Leu Ile              20 25 30 Tyr Pro Ile Phe Val Thr Asp Val Pro Asp Asp Val Gln Pro Ile Ala          35 40 45 Ser Leu Pro Gly Val Ala Arg Tyr Gly Val Asn Gln Leu Glu Glu Met      50 55 60 Leu Arg Pro Leu Val Glu Ala Gly Leu Arg Cys Val Leu Ile Phe Gly  65 70 75 80 Val Pro Ser Arg Val Pro Lys Asp Glu Gln Gly Ser Ala Ala Asp Ser                  85 90 95 Glu Asp Ser Pro Thr Ile Glu Ala Val Arg Leu Leu Arg Lys Thr Phe             100 105 110 Pro Ser Leu Leu Val Ala Cys Asp Val Cys Leu Cys Pro Tyr Thr Ser         115 120 125 His Gly His Cys Gly Leu Leu Ser Glu Asn Gly Ala Phe Leu Ala Glu     130 135 140 Glu Ser Arg Gln Arg Leu Ala Glu Val Ala Leu Ala Tyr Ala Lys Ala 145 150 155 160 Gly Cys Gln Val Val Ala Pro Ser Asp Met Met Asp Gly Arg Val Glu                 165 170 175 Ala Ile Lys Ala Leu Leu Lys His Gly Leu Gly Asn Arg Val Ser             180 185 190 Val Met Ser Tyr Ser Ala Lys Phe Ala Ser Cys Phe Tyr Gly Pro Phe         195 200 205 Arg Asp Ala Gln Ser Ser Pro Ala Phe Gly Asp Arg Arg Cys Tyr     210 215 220 Gln Leu Pro Pro Gly Ala Arg Gly Leu Ala Leu Arg Ala Val Ala Arg 225 230 235 240 Asp Ile Gln Glu Gly Ala Asp Met Leu Met Val Lys Pro Gly Leu Pro                 245 250 255 Tyr Leu Asp Met Val Glu Val Lys Asp Lys His Pro Glu Leu Pro             260 265 270 Leu Ala Val Tyr Gln Val Ser Gly Glu Phe Ala Met Leu Trp His Gly         275 280 285 Ala Gln Ala Gly Ala Phe Asp Leu Arg Thr Ala Val Leu Glu Thr Met     290 295 300 Thr Ala Phe Arg Ala Gly Ala Asp Ile Ile Ile Thr Tyr Phe Ala 305 310 315 320 Pro Gln Leu Leu Lys Trp Leu Lys Glu Glu                 325 330 <210> 10 <211> 361 <212> PRT <213> Artificial Sequence <220> <223> L-lactate dehydrogenase <400> 10 Met Ser Lys Ser Ser Gly Gly Tyr Thr Tyr Thr Glu Thr Ser Val Leu   1 5 10 15 Phe Phe His Phe Lys Val Ser Lys Asp Ser Lys Ser Lys Met Ala Thr              20 25 30 Leu Lys Asp Gln Leu Ile Val Asn Leu Leu Lys Glu Glu Gln Ala Pro          35 40 45 Gln Asn Lys Ile Thr Val Val Gly Val Gly Ala Val Gly Met Ala Cys      50 55 60 Ala Ile Ser Ile Leu Ale Leu Ale Leu  65 70 75 80 Asp Val Met Glu Asp Lys Leu Lys Gly Glu Met Met Asp Leu Gln His                  85 90 95 Gly Ser Leu Phe Leu Lys Thr Pro Lys Ile Val Ser Ser Lys Asp Tyr             100 105 110 Cys Val Thr Ala Asn Ser Lys Leu Val Ile Ile Thr Ala Gly Ala Arg         115 120 125 Gln Gln Glu Gly Glu Ser Arg Leu Asn Leu Val Gln Arg Asn Val Asn     130 135 140 Ile Phe Lys Phe Ile Ile Pro Asn Ile Val Lys Tyr Ser Pro His Cys 145 150 155 160 Lys Leu Leu Ile Val Ser Asn Pro Val Asp Ile Leu Thr Tyr Val Ala                 165 170 175 Trp Lys Ile Ser Gly Phe Pro Lys Asn Arg Val Ile Gly Ser Gly Cys             180 185 190 Asn Leu Asp Ser Ala Arg Phe Arg Tyr Leu Met Gly Glu Arg Leu Gly         195 200 205 Val His Ala Leu Ser Cys His Gly Trp Val Leu Gly Glu His Gly Asp     210 215 220 Ser Ser Val Ser Val Ser Val Ser Val Ser Val Ser Val 225 230 235 240 Lys Ser Leu Asn Pro Glu Leu Gly Thr Asp Ala Asp Lys Glu Gln Trp                 245 250 255 Lys Glu Val His Lys Gln Val Val Asp Ser Ala Tyr Glu Val Ile Lys             260 265 270 Leu Lys Gly Tyr Thr Ser Trp Ala Ile Gly Leu Ser Val Ala Asp Leu         275 280 285 Ala Glu Ser Ile Met Lys Asn Leu Arg Arg Val Val Pro Ile Ser Thr     290 295 300 Met Ile Lys Gly Leu Tyr Gly Ile Asn Glu Asp Val Phe Leu Ser Val 305 310 315 320 Pro Cys Ile Leu Gly Gln Asn Gly Ile Ser Asp Val Val Lys Val Thr                 325 330 335 Leu Thr Pro Glu Glu Glu Ala Arg Leu Lys Lys Ser Ala Asp Thr Leu             340 345 350 Trp Gly Ile Gln Lys Glu Leu Gln Phe         355 360 <210> 11 <211> 539 <212> PRT <213> Artificial Sequence <220> <223> T-complex protein 1 subunit delta <400> 11 Met Pro Glu Asn Val Ala Ser Arg Ser Gly Ala Pro Thr Ala Gly Pro   1 5 10 15 Gly Ser Arg Gly Lys Ser Ala Tyr Gln Asp Arg Asp Lys Pro Ala Gln              20 25 30 Ile Arg Phe Ser Asn Ile Ser Ala Ala Lys Ala Val Ala Asp Ala Ile          35 40 45 Arg Thr Ser Leu Gly Pro Lys Gly Met Asp Lys Met Ile Gln Asp Gly      50 55 60 Lys Gly Asp Val Thr Ile Thr Asn Gly Ala Thr Ile Leu Lys Gln  65 70 75 80 Met Gln Val Leu His Pro Ala Ala Arg Met Leu Val Glu Leu Ser Lys                  85 90 95 Ala Gln Asp Ile Glu Ala Gly Asp Gly Thr Thr Ser Val Val Ile Ile             100 105 110 Ala Gly Ser Leu Leu Asp Ser Cys Thr Lys Leu Leu Gln Lys Gly Ile         115 120 125 His Pro Thr Ile Ile Ser Glu Ser Phe Gln Lys Ala Leu Glu Lys Gly     130 135 140 Leu Glu Ile Leu Thr Asp Met Ser Arg Pro Val Gln Leu Ser Asp Arg 145 150 155 160 Glu Thr Leu Leu Asn Ser Ala Thr Thr Ser Leu Asn Ser Lys Val Val                 165 170 175 Ser Gln Tyr Ser Ser Leu Leu Ser Pro Met Ser Val Asn Ala Val Met             180 185 190 Lys Val Ile Asp Pro Ala Thr Ala Thr Ser Val Asp Leu Arg Asp Ile         195 200 205 Lys Ile Val Lys Lys Leu Gly Gly Thr Ile Asp Asp Cys Glu Leu Val     210 215 220 Glu Gly Leu Val Leu Thr Gln Lys Val Ala Asn Ser Gly Ile Thr Arg 225 230 235 240 Val Glu Lys Ala Lys Ile Gly Leu Ile Gln Phe Cys Leu Ser Ala Pro                 245 250 255 Lys Thr Asp Met Asp Asn Gln Ile Val Val Ser Asp Tyr Ala Gln Met             260 265 270 Asp Arg Val Leu Arg Glu Glu Arg Ala Tyr Ile Leu Asn Leu Val Lys         275 280 285 Gln Ile Lys Lys Thr Gly Cys Asn Val Leu Leu Ile Gln Lys Ser Ile     290 295 300 Leu Arg Asp Ala Leu Ser Asp Leu Ala Leu His Phe Leu Asn Lys Met 305 310 315 320 Lys Ile Met Val Val Lys Asp Val Glu Arg Glu Asp Ile Glu Phe Ile                 325 330 335 Cys Lys Thr Ile Gly Thr Lys Pro Val Ala His Ile Asp Gln Phe Thr             340 345 350 Ala Asp Met Leu Gly Ser Ala Glu Leu Ala Glu Glu Val Ser Leu Asn         355 360 365 Gly Ser Gly Lys Leu Phe Lys Ile Thr Gly Cys Thr Ser Pro Gly Lys     370 375 380 Thr Val Thr Ile Val Val Arg Gly Ser Asn Lys Leu Val Ile Glu Glu 385 390 395 400 Ala Glu Arg Ser Ile His Asp Ala Leu Cys Val Ile Arg Cys Leu Val                 405 410 415 Lys Lys Arg Ala Leu Ile Ala Gly Gly Gly Ala Pro Glu Ile Glu Leu             420 425 430 Ala Leu Arg Leu Thr Glu Tyr Ser Arg Thr Leu Ser Gly Met Glu Ser         435 440 445 Tyr Cys Val Ala Phe Ala Asp Ala Met Glu Val Ile Pro Ser Thr     450 455 460 Leu Ala Glu Asn Ala Gly Leu Asn Pro Ile Ser Thr Val Thr Glu Leu 465 470 475 480 Arg Asn Arg His Ala Gln Gly Glu Lys Thr Thr Gly Ile Asn Val Arg                 485 490 495 Lys Gly Gly Ile Ser Asn Ile Leu Glu Glu Met Val Val Gln Pro Leu             500 505 510 Leu Val Ser Ser Ala Leu Thr Leu Ala Thr Glu Thr Val Arg Ser         515 520 525 Ile Leu Lys Ile Asp Asp Val Val Asn Thr Arg     530 535 <210> 12 <211> 198 <212> PRT <213> Artificial Sequence <220> <223> peroxiredoxin 2 <400> 12 Met Ala Ser Gly Asn Ala Gln Ile Gly Lys Ser Ala Pro Asp Phe Thr   1 5 10 15 Ala Thr Ala Val Val Asp Gly Ala Phe Lys Glu Ile Lys Leu Ser Asp              20 25 30 Tyr Arg Gly Lys Tyr Val Val Leu Phe Phe Tyr Pro Leu Asp Phe Thr          35 40 45 Phe Val Cys Pro Thr Glu Ile Ile Ala Phe Ser Asp His Ala Glu Asp      50 55 60 Phe Arg Lys Leu Gly Cys Glu Val Leu Gly Val Ser Val Asp Ser Gln  65 70 75 80 Phe Thr His Leu Ala Trp Ile Asn Thr Pro Arg Lys Glu Gly Gly Leu                  85 90 95 Gly Pro Leu Asn Ile Pro Leu Leu Ala Asp Val Thr Lys Ser Leu Ser             100 105 110 Gln Asn Tyr Gly Val Leu Lys Asn Asp Glu Gly Ile Ala Tyr Arg Gly         115 120 125 Leu Phe Ile Ile Asp Ala Lys Gly Val Leu Arg Gln Ile Thr Val Asn     130 135 140 Asp Leu Pro Val Gly Arg Ser Val Asp Glu Ala Leu Arg Leu Val Gln 145 150 155 160 Ala Phe Gln Tyr Thr Asp Glu His Gly Glu Val Cys Pro Ala Gly Trp                 165 170 175 Lys Pro Gly Ser Asp Thr Ile Lys Pro Asn Val Asp Asp Ser Lys Glu             180 185 190 Tyr Phe Ser Lys His Asn         195 <210> 13 <211> 2104 <212> DNA <213> Artificial Sequence <220> <223> Heat shock cognate 71 kDa protein <400> 13 ggtctcattg aacgcggagg cagctgcctg gcatttgtgt ggtctcgtcg tcagcgcagc 60 tgggcctaca cacaagcaac catgtctaag ggacctgcag ttggcattga tctcggcacc 120 acctactcct gtgtgggtgt cttccagcat ggaaaggtgg aaattattgc caatgaccag 180 ggtaaccgca ccacgccaag ctatgttgct ttcacggaca cagagagatt aattggggat 240 gcggccaaga atcaggttgc aatgaacccc accaacacag tttttgatgc caaacgtctg 300 atcgggcgta ggtttgatga tgctgttgtt cagtctgata tgaagcactg gcccttcatg 360 gtggtgaatg atgcaggcag gcccaaggtc caagtggaat acaaagggga gacaaaaagt 420 ttctacccag aggaagtgtc ctccatggtt ctgacaaaga tgaaggaaat tgcagaagcg 480 tacctcggaa agaccgttac caacgctgtg gtcacagtgc ccgcttactt caatgactct 540 cagcgacagg caacaaaaga tgctggaact attgctggcc tcaatgtact tcgaatcatc 600 aatgaaccaa ctgctgctgc tattgcttac ggcttagata agaaggtcgg agctgaaagg 660 aatgtgctca tttttgactt gggaggtggc acttttgatg tgtcaatcct cactattgag 720 gatggaattt ttgaggtcaa atcaacagct ggagacaccc acttaggtgg agaagatttt 780 gacaaccgaa tggtcaatca tttcattgct gagttcaagc gaaagcacaa gaaagacatc 840 agtgagaaca agagagctgt ccgccgtctc cgcacggcct gcgagcgggc caagcgcacc 900 ctctcctcca gcacccaggc cagtattgag attgattctc tctatgaggg aattgacttc 960 tatacctcca ttacccgggc tcgatttgag gagttgaatg ctgacctgtt ccgtggcaca 1020 ctggaccctg tagagaaggc ccttcgagat gccaagctgg acaagtcaca gatccatgat 1080 attgtcttgg tgggtggttc taccagaatc cccaagattc agaaacttct gcaagacttc 1140 ttcaatggaa aagagctgaa caagagcatt aaccccgatg aagctgttgc ctatggtgca 1200 gctgtccagg cagccattct atctggagac aagtctgaga acgttcagga tttgctgctc 1260 ttggatgtca ctcctctttc ccttggtatt gaaactgctg gcggagtcat gactgtcctc 1320 atcacccca ataccaccat ccccaccaag cagacacaga ctttcaccac ctactctgac 1380 aaccagcctg gtgtactcat tcaggtgtat gaaggtgaaa gggccatgac caaggacaac 1440 aacctgcttg gaaagttcga gctcacaggc atccctccag caccccgtgg ggtccctcag 1500 attgaggtta cttttgacat cgatgccaat ggcatcctca atgtttctgc tgtagataag 1560 agcacaggaa aggagaacaa gatcaccatc accaatgaca agggccgctt gagtaaggaa 1620 gatattgagc gcatggtcca agaagctgag aagtacaagg ctgaggatga gaagcagaga 1680 gataaggttt cctccaagaa ctcactggag tcctatgcct tcaacatgaa agcaactgtg 1740 gaagatgaga aacttcaagg caagatcaat gatgaggaca aacagaagat tcttgacaag 1800 tgcaatgaaa tcatcagctg gctggataag aaccagactg cagagaagga agaatttgag 1860 catcagcaga aagaactgga gaaagtctgc aaccctatca ttaccaagct gtaccagagt 1920 gcaggtggca tgcctggggg aatgcctggt ggcttcccag gtggaggagc tcccccatct 1980 ggtggtgctt cttcaggccc caccattgaa gaggtggatt aagtcagtcc aagaaggtgt 2040 agctttgttc cacagggacc caaaacaagt aacatggaat aataaaacta tttaaattgg 2100 cacc 2104 <210> 14 <211> 3139 <212> DNA <213> Artificial Sequence <220> <223> Inter-alpha trypsin inhibitor, heavy chain 2 <400> 14 ttctttttaa aacgatctgc tctcatcaac agacaagttc ctgctaacct ggttccgtag 60 gtggggagtt ctccccagac catctgctcg ggggagcttg gcacagatgt ccagcaaaat 120 gcagcggcct gtgtgccttc tcatctggct gtttcttctg gaagcacaag ccttcgaaat 180 ccccataaac ggaaactcag aatttgcaga atacagtgat cttgtggaac tggccccaga 240 taaactccca tttgtgcaag agaatggaag acatcagaga agccttcctg aggaatccgg 300 agaggagacg gacactgttg atccagtgac tctttatagc tacaaagtcc agtccactat 360 tacttctcgg gtggccacaa ctaccatcca gagcaaactg gtgaacaatt ccccattgcc 420 tcagagtgtc gtgttcgatg ttcagattcc caaaggagcc tttatctcca acttcaccat 480 gactgtgaac ggcatgacat ttacaagctc cattaaagag aaaactgtgg gccgagctct 540 ttattcacag gcaagagcaa agggcaagac ggctggatgg gtaaggagca gaactctgga 600 tatggagaac ttcaacacag aggtgaacat cccgcctgga gccaaggtgc agtttgaact 660 tcattaccag gaggtgaagt ggaggaagct gggatcctat gagcacaaga ttcatttgca 720 gcccgggaag ctggccaaac acttggaggt gaatgtgtgg attatcgaac cccaaggaat 780 gagatttctt catgttcctg acacatttga aggccatttc caaggcgtcc ctgtcatatc 840 taaaggacaa cagaaggcac atgtatcctt caagcccaca gtagcacaac aaagaaaatg 900 ccccaactgc acagagactg cagtgaacgg ggagctggtg gtgatgtatg atgtcaacag 960 agaagagaag gccggggaac ttgaggtgtt taatggatat tttgttcact tctttgctcc 1020 tgagaacctg gacccaatcc ccaagaacat cctttttgtt attgacgtca gtggctccat 1080 gtgggggatt aagatgaagc agactgtgga ggcaatgaag acgatattgg atgacctaag 1140 aacagacgac caattctctg tggttgattt caaccataat gttcgaacct ggagaaatga 1200 tttagtttca gctactaaaa cacaaattgc agatgccaag agatacattg agaaaattca 1260 gcctagtgga ggcacgaata tcaacgaggc actgcttcga gcaattttca ttttgaatga 1320 agccagtaac atgggattgc tgaaccccga ctcagtctct ctgatcattt tggtttctga 1380 tggagatcca accgtagggg aactaaaatt gtccaaaatt cagaaaaatg tgaagcagag 1440 catccaagat aacatctccc tgtttagttt gggaatcgga tttgatgtcg actatgattt 1500 tttgaagagg ctgtccaatg aaaatcgtgg tattgctcaa cggatctacg ggaaccagga 1560 cacatcctct cagctcaaga aattttacaa ccaggtctct actccactgc tcagaaatgt 1620 tcagtttaac tacccgcagg catcagtaac agatgtcacc caaaacaatt tccacaacta 1680 cttcggaggt tctgagatcg tggtggcagg gaaatttgat cccagcaaac tgacagaagt 1740 ccagagcatc atcacggcaa cttcggctaa cacagaattg gtcttggaaa ccctgagcca 1800 gatggatgac ttggaggagt ttctgtcaaa ggacaagcat gcagaccctg acttcaccaa 1860 gaagctgtgg gcctatctca ccatcaacca actactagca gagagaagtc tggctcctac 1920 tgctgccatc aaaaggaaaa tcaccaaaac aatcctgcag atgtctctag accatcacat 1980 cgtgactccg cttactgcca tggtgataga gaatgatgct ggagatgaac gcatgctggc 2040 tgactccccg ccacaggacc actcctgctg ctcaggcgca ctgtattatg gcaccaaggt 2100 tgcctcgggt cccatcccat catgggccaa cccatctccg acaccaatgt cggccatgct 2160 tgcagtcgga gcgaagccac ttgagtctac tccacctact catttgaatc aagtggaaaa 2220 tgacccccac ttcatcattt acctgcccaa aagcaaaagg aatatttgtt tcaatattga 2280 ctcagaacct gggaaaatcc taagcctggt gtctgaccca gaatcaggga ttgtagtcaa 2340 cggtcagctt atcggcgcca agagggcaga gaatggaaag ctaagcacct actttggaaa 2400 actggggttt tatttccaaa aggaaggcat gaagatagaa atcagcaccg agaccatcac 2460 cctgagctct ggctcctcta catctcgact gtcctggtcc gacacagctc atcttggaaa 2520 ctcgagggtg ctcatctctg tgaagaaaga aaaatctgtg actctcaccc tgaataaaga 2580 gctgttcttt tccgttctgc tgcatcgtgt gtggaggaag cacccagtta acgtggactt 2640 cctggggatc tacgctcccc ccatagacaa gttctcacct agagtgcatg gactgttagg 2700 ccagttcatg caggagccag ccatccacat cttcaatgag agacccggaa aggagccggg 2760 aaaacccgag gcaagcatgg aggtgaaagg acataagctg actgtcacca gaggcctgca 2820 gaaggactac aggacggata tcgtgtttgg aacagacgtt ccctgctggt ttgtacacaa 2880 cagtgggaaa ggtttcatcg atggacatta taaggattat tttgtgcctc agctctacag 2940 ctttctcaag cggccttaat ggcttatagt tttggaaact gtatgtgtat ccttttctcc 3000 cttgatactt tttgcagtta ctcccccgtt tcagcaactc aaaataaacg cagatattat 3060 agtggcctaa aaggcctgct aacccaccgg agaaaaataa atatttgcaa aaaaggctca 3120 ggaaaaaaaa aaaaaaaaa 3139 <210> 15 <211> 1812 <212> DNA <213> Artificial Sequence <220> <223> Vitamin D-binding protein <400> 15 tttgccttat gatgatatta ctatctattt ttgggaacaa ggctgattgt cccttgcttc 60 tgggcagaga ttaataattg atgagtttct agttaggatc gtttgcaaat cctgtgctcc 120 tgccttttgc gtggtggccg gcaggtgaga ggaggtgctg caagactctc tggtcgcaga 180 atgaagaggg ttctggttct cctgctggcc ttagcctttg ggcacgctct agagagaggc 240 cgagactatg agaaggataa agtttgcaat gaactcgcca tgctggggaa agaggacttc 300 agatctttat cactaatcct atatagcagg aagttttcca gtagcacatt tgagcaggtc 360 aaccagcttg tgaaggaagt tgtctccttg actgaggagt gctgtgctga gggagctgac 420 cccacctgct acgacaccag gacctcagag ctgtctgtta agtcctgtga aagtgatgct 480 ccctttccgg ttcaccctgg aactcctgag tgttgcacca aggaggggct ggagcggaaa 540 ctctgcatgg ctgccctcag tcaccagccg caggaatttc ccacctacgt ggaaccaaca 600 aatgatgaga tctgtgaggc cttcaggagg gacccaaagg gatttgctga ccagtttctg 660 tatgagtatt ccagcaatta tggacaagcg cctctgccac ttttagttgc ttacaccaag 720 aattatctct ctatggttgg atcctgctgt acttctgcaa acccaactgt atgctttgtg 780 aaggagagac tccagatgaa gcatttgtca cttctcacca ccatgtcaaa cagagtctgc 840 tcacaatatg ctgcatatgg aaaggaaaaa tcaaggctga gccatctcat aaaactagcc 900 caaaaagtgc caactgctaa cctggagaac gttctgccac tagctgaaga ctttactgaa 960 atcctgtcca gatgttgtga gtctacctca gaggattgca tggccagtga gctgcctgag 1020 cacacaataa aaatctgtca aaacttatcc aaaaagaatt ctaagtttga agagtgctgt 1080 caagaaaaca cacccatgaa catttttatg tgcacctact tcatgccagc tgctgaacca 1140 cttcaattgc cagctatcaa gttgccaact ggcaaagacc tctgtggtca gagtaccaca 1200 caagccatgg accagtatac atttgaacta agcagaagga ctcaagttcc agaagtgttc 1260 ctcagcaaag ttctggagcc aaccctgaaa acccttaggg agtgctgtga cactcaggat 1320 tctgttgcct gtttcagcac tcagagtccc ctgctgaaga ggcaactaac ttctttcatc 1380 gaaaaaggtc aagaaatgtg tgcagattat tctgagaaca catttactga gtacaagaaa 1440 aaattggcag aacggctaag gacaaaaaca cccaacacct ctccggcaga gctgaaagac 1500 atggtggaga aacactcgga ctttgcctct aagtgctgct ctataaactc acctcctctc 1560 tactgcagct cacagattga tgcagaaatg atagacaccc tgcagtcctg atcagggccc 1620 gtgcactagc ttggatcttg aactgaacca ctctggaaaa ttgccactgg taacaaaatc 1680 aagcaccaca gagatggcct tccaagaaga tcaccaagat aattccattt ttcttagcta 1740 caatgttttt ggagaattat aaaaaataaa taaataaatt gaaatatggt gcaaactcta 1800 attatgatgc ag 1812 <210> 16 <211> 8560 <212> DNA <213> Artificial Sequence <220> <223> Talin-1 <400> 16 aactcggaag tggctcctgg gctgcgccac gtcccggggg ctatgcaaat tatggggacg 60 tcctttcaag cttcccacgc cccagggcgg ggattctgag gatccttgtc cgccttcgcc 120 tcgagctatt aaaaaaaaaa acttaagcgt ttaaaaggga actccagctt agttcgggcg 180 ttctctccag agctctgcac gctcgcttcc tgctggggag gggcggccag gcttcgcggg 240 cgcccgagca tcgagaacta cggccagagc agcttcctgc agtctccgct accatagagc 300 gcgggcccag ggcgccgccg gcgggtgggg gacgtttcca ggacggaagt ggccgagagt 360 gtgtcgaagg gagggcgagg ccggagcccc aaaagcgacc gggagaagga gcgggtagcg 420 ggcccagggc ggggcgcaga gccgggcagc gcaggtatcg ccaggccgaa gagcagaagc 480 tgccaccatg gttgcgcttt cgctgaagat tagcattggg aatgtggtga agacgatgca 540 atttgagcca tctaccatgg tgtacgacgc ctgccgcatg attcgtgagc ggatcccaga 600 ggccctggct ggccctccca acgactttgg gctctttctg tcagatgatg accccaaaaa 660 aggcatctgg ctagaggctg ggaaagcttt ggactactac atgctccgaa atggggacac 720 catggagtac agaaaaaagc agagacccct gaagatccgg atgttagacg gaacggtgaa 780 gactatcatg gtggatgact ccaagaccgt cacagacatg ctcatgacca tttgtgcccg 840 aattggtatc accaaccatg atgagtattc actggttcga gagctgatgg aagaaaagaa 900 agatgagggg acagggaccc tgagaaaaga caagacccta ctgcgagatg aaaagaagat 960 ggagaagcta aagcagaagt tgcacacaga cgatgagttg aactggctgg accatgggcg 1020 gacactgagg gaacagggag tggaagagca tgagacgttg ctgctgcgga gaaagttctt 1080 ctactcagac cagaatgtgg attcccgaga ccctgtacag ctgaaccttc tctatgtgca 1140 ggcacgagat gacatcctga atggctccca tcctgtctcc tttgacaagg cctgtgaatt 1200 tgcaggcttc cagtgccaga tccagtttgg acctcacaat gaacagaagc acaaggctgg 1260 cttccttgac ctgaaggact tcctgcccaa ggagtatgtg aagcagaagg gagagcgtaa 1320 gatctttcag gcacacaaga attgtgggca gatgagtgaa attgaggcca aggtacgcta 1380 cgtgaagctg gcccgttccc tcaagactta cggtgtctcc ttcttcctag tcaaggaaaa 1440 gatgaagggg aagaataaac tggtgcccag gctgttgggc atcactaagg agtgtgtgat 1500 gcgtgtggat gagaagacca aggaggtgat tcaggaatgg agcctcacca acatcaaacg 1560 ctgggctgcc tctcccaaga gctttactct ggactttgga gactatcagg atggctacta 1620 ctcagtacag acgacagaag gcgagcagat cgcacagctc attgctggct acatagatat 1680 catccttaag aagaaaaaaa gcaaggacca ttttgggctg gagggagatg aagagtctac 1740 tatgttggag gactcagtgt cacccaaaaa gtcaacagtc cttcagcagc agtacaaccg 1800 agtggggaaa gtggagcacg gctctgtggc tctgccagcc atcatgcgct ctggagcctc 1860 tggtcctgag aatttccagg tggggagcat gccacctgcc cagcagcaga tcaccagtgg 1920 ccagatgcac cgaggacaca tgccacctct gacttcagcc cagcaggcgc tcactggaac 1980 cattaactcc agcatgcagg ctgtgcaggc tgcccaggcc actctggatg actttgagac 2040 tctaccccct cttggccagg atgctgcctc caaagcttgg cgtaagaaca agatggatga 2100 atcaaagcat gagatccact cccaggtaga tgccatcaca gctggcactg cctctgtggt 2160 aaacctgaca gcaggagatc ctgcagagac agactataca gcagtgggct gtgcagtcac 2220 caccatctct tccaacctga cggagatgtc ccgtggggtg aagctattgg ctgccctgct 2280 ggaggatgaa ggcggcaatg gccggcccct tctgcaagca gcaaagggcc ttgcaggggc 2340 tgtgtcggaa ctgcttcgca gtgctcagcc cgccagcgct gagccccgtc agaacctgct 2400 gcaggcagcc gggaacgtgg gccaggccag tggggagctg ttgcagcaaa ttggggaaag 2460 tgacactgac ccccacttcc aggatgttct aatgcagcta gcaaaggcag tggcaagtgc 2520 tgcagccgcc ctggtcctca aggccaagag tgtggcccag cgcacagagg attccgggct 2580 tcagacccaa gtgattgctg cagctacgca atgtgctctg tccacttccc agctggtggc 2640 ctgtaccaag gtggtagcac ctacaatcag ctcacctgtg tgccaagaac agctagtcga 2700 ggctgggcga ctggtggcca aagctgtgga gggctgtgtg tctgcctcgc aagcagctac 2760 agaagatgga cagctgcttc gaggggtagg agcagcagcc acggctgtca cccaggccct 2820 caatgagctg ctgcagcacg tgaaggccca cgccaccgga gctgggcctg ccggtcggta 2880 tgaccaagcc actgatacca tcctcaccgt cactgaaaac atcttcagct ccatgggtga 2940 tgctggggag atggtgcgcc aggcccgcat cctggcccaa gccacctcag acctggtcaa 3000 tgctatcaag gccgatgctg agggggagag tgatctggag aactctagaa agctcctgag 3060 tgctgccaag atcctcgctg atgccaccgc caagatggtg gaggcggcca agggagcagc 3120 cgcccaccct gacagtgagg aacagcagca gcgactgcgt gaagcagctg aggggcttcg 3180 catggccacc aatgcagctg cgcagaacgc catcaagaag aagttggtgc agcgcctgga 3240 gcatgcagcc aagcaagctg cagcctctgc aacacagacc attgctgcag cccaacatgc 3300 agcctctgcc cccaaggcct ctgcgggccc ccagccccta ctggtgcaga gctgtaaggc 3360 cgtggcagag cagattccgt tgctggtgca gggtgtccga ggaagtcaag ctcagcctga 3420 cagccctagt gctcagctcg ccctcattgc tgccagccag agcttcctgc agccaggtgg 3480 gaagatggtg gcagcggcaa aagcctcagt gccaacaatt caggaccagg cttcagcaat 3540 gcagctgagt cagtgcgcaa agaacctagg cactgccctg gccgagctgc gcactgctgc 3600 tcagaaggct caggaggcat gtggaccttt ggagatggat tctgcactga gtgttgtaca 3660 aaatctagag aaagacctac aggaaataaa agcagcagct cgagatggca agcttaagcc 3720 cttacccggg gaaacaatgg agaagtgtac ccaagatctt ggcaacagca ccaaagcggt 3780 gagctctgcc atcgccaagc tgctggggga gattgcccag ggcaatgaga actatgcagg 3840 tattgcagct cgggatgtag ctggtggact aaggtcacta gcccaggctg cgcggggtgt 3900 ggctgcgctg acatcagatc ctgcagtgca ggccattgtg cttgacacag ccagcgacgt 3960 tctggacaag gccagcagcc tcattgaaga ggcaaaaaaa gcatctggac acccagggga 4020 cccagaaagc cagcaaaggc ttgctcaggt agctaaagca gtgacccaag ccctgaaccg 4080 ctgtgtcagc tgcctgcctg gccaaagaga cgtggataat gccctacgag ccgttggaga 4140 tgccagcaag cgactcctga gtgactcgct tcctccaagc acggggacat ttcaagaagc 4200 acagagccga ttgaatgaag ctgcagctgg gttaaatcag gccgccacag aactggtgca 4260 agcctcccga ggaactcctc aggacctggc tcgggcctcc ggccgatttg gacaggactt 4320 cagcaccttc ctggaagctg gtgtggagat ggccggacag gccccgagcc aggaggaccg 4380 agcccaggtg gtgtccaacc tgaagggcat atccatgtct tcaagcaaac ttctccttgc 4440 cgctaaggcc ttgtccacag accctgcttc tcccaacctc aagagtcagc tggctgcggc 4500 tgcccgggca gtgacggaca gcatcaacca gctcatcacc atgtgcaccc agcaggcacc 4560 tggccagaag gagtgtgaca atgcacttcg gcagctagag acagtccgag aactcctgga 4620 gaatccagtc cagcccatca acgacatgtc ctacttcggt tgcttggaca gtgtcatgga 4680 gaactccaag gtcctaggtg aggccatgac tggcatctcc caaaatgcca agaatggaaa 4740 tctgccggag tttggagatg ccattgccac agcctccaag gctctctgtg gcttcacgga 4800 ggcagccgca caggcagcat atctagttgg tgtctctgac cccaacagcc aagctggaca 4860 gcaaggactg gtggaaccca cacagtttgc ccgtgcaaac caggcaattc agatggcctg 4920 tcagagtctg ggggagcctg gctgtaccca ggcccaggtg ttatctgcag ccactattgt 4980 agccaaacac acatctgcat tgtgtaacag ctgtcgcctg gcttccgcta ggactgccaa 5040 tcccactgcc aagcgccagt ttgtacagtc agccaaggag gtggccaaca gtacagccaa 5100 tcttgtcaag accatcaagg cactagatgg ggacttcaca gaagagaacc gtgcccagtg 5160 ccgagcagcc acagcccctc tgctggaagc tgtggataac ctgagtgcct ttgcctccaa 5220 ccctgagttc tccagcgtcc ctgcccagat cagccctgag ggcagagcag ccatggagcc 5280 cattgtaatc tctgctaaga caatgttgga gagtgctgga gggctcatcc agacagctcg 5340 ggcgttagca gtcaatcccc gagacccccc acgttggtct gtgctagctg gccactctcg 5400 gactgtctca gattccatca agaaacttat tacaagcatg agggacaaag ccccagggca 5460 gctggagtgt gagacagcca ttgcggctct gaacagctgc ttgcgggacc tagatcaggc 5520 ttcgcttgcc gctgtcagcc agcagcttgc tccccgtgaa ggaatctctc aagaggcttt 5580 gcacacccag atgctcactg cagtgcaaga aatttctcat ctcattgagc cgctggccag 5640 cgctgctcgg gctgaagcct cccagctggg acacaaggtg tcccaaatgg cccagtactt 5700 tgaaccactc accctggctg cagtgggtgc tgcgtctaag accctgagcc acccacagca 5760 gatggcactt ctggaccaga ctaaaacgtt ggcagagtct gccttgcagt tgctatatac 5820 tgccaaggag gctggtggta accccaagca agcagcacac acccaggaag ccctggagga 5880 ggctgtgcag atgatgacag aggccgtaga agacctgacg acgaccctca acgaagcagc 5940 cagtgctgca ggagtcgttg gcggcatggt ggactctatc actcaggcca tcaaccagct 6000 agatgaagga cctatgggtg acccagaagg ctcattcgta gattaccaga caaccatggt 6060 gaggacagcc aaggccattg ctgtcactgt tcaggagatg gtaaccaagt caaacaccag 6120 ccctgaagag ctaggccctc ttgccaacca gctgaccagt gactatggcc gactggcctc 6180 acaagccaag cctgcagctg tggctgctga aaatgaagag ataggcgctc acatcaaaca 6240 ccgagtacag gagctgggcc atggctgctc tgctctggtc accaaggcag gtgccttgca 6300 gtgtagcccc agtgatgtct acaccaagaa ggagctcata gagtgtgccc gcagagtgtc 6360 agagaaggtc tcccatgttc tggctgcact ccaggctggg aatcgtggta cccaggcctg 6420 cattacagca gccagtgctg tgtctggtat cattgctgac ctcgacacca ccatcatgtt 6480 cgctactgct ggcacactta accgtgaggg tgctgaaact tttgctgacc accgggaggg 6540 tatcttaaag acagcaaagg ttcttgtgga ggacaccaag gtcctagtgc agaatgcagc 6600 tgggagccag gagaagttgg cacaagccgc ccagtcctcc gtggccacca ttacccgcct 6660 cgctgatgtg gtcaagctcg gtgcagccag cctaggagcc gaagaccctg aaactcaggt 6720 ggtgctgatc aatgcagtaa aggacgtagc caaggccctg ggtgacctca tcagcgctac 6780 gaaggctgca gcgggcaaag ttggggatga ccctgcagtg tggcagctca agaactctgc 6840 caaggtgatg gtgaccaatg tgacatcatt gctcaagaca gtgaaggctg tggaagatga 6900 ggccaccaaa ggcacacggg ccctagaggc aaccacagag cacatacgtc aggaactggc 6960 ggtcttctgt tccccagagc cacctgccaa gacctctacc cctgaagact tcatccgaat 7020 gaccaagggt attactatgg caacagccaa agccgttgct gctggcaatt cctgtcgaca 7080 ggaagatgtc attgccacag ccaatctgag ccgacgggct attgcggaca tgcttcgggc 7140 ttgcaaggaa gcagctttcc acccagaagt ggcgcctgat gtacggctcc gagccctgca 7200 ctatggccgg gagtgtgcca atggttacct ggaactgctg gaccacgtgc tgctgaccct 7260 tcagaagcca aacccagacc tgaagcagca gctaactgga cactcaaagc gggttgctgg 7320 ctctgtgact gagctcatcc aggctgctga agccatgaaa ggaacagagt gggtggaccc 7380 agaggaccct actgtcattg ctgagaatga actcttggga gccgcagccg ccatcgaggc 7440 tgcagccaag aagctggagc agctgaagcc cagggccaaa cccaaggagg cggatgagtc 7500 cttgaacttt gaggaacaaa tcctagaagc tgccaagtcc atcgctgcag ccaccagtgc 7560 actggtaaag gctgcgtcag cagcccagag ggagctggtg gctcaaggaa aggtgggcgc 7620 cattccagcc aatgcactgg atgatgggca gtggtcgcag ggcctcattt ccgctgcccg 7680 tatggtggct gcggccacca acaatctgtg tgaagcagct aacgcagctg tccagggcca 7740 tgctagccag gagaaactca tttcctcagc caagcaagta gctgcctcca cagctcagct 7800 cctggtagct tgcaaggtca aagcagatca ggactctgag gcaatgaaac ggctccaggc 7860 tgctggcaat gcagtgaaga gggcctcaga taacctggta aaggcggccc agaaagctgc 7920 agccttcgaa gaccaggaga atgagacggt ggtggtgaag gagaaaatgg ttgggggcat 7980 tgcccagatt atcgcagcac aggaagagat gcttcggaag gaacgagagc tggaagaggc 8040 tcggaaaaag ctcgcccaga tccggcagca gcagtacaag ttcttgcctt cagagcttcg 8100 agacgagcac taaaaaagcc ctgtgtattt aatgcagacc cagcccagag actgtgcctg 8160 ccactaccaa agccttctgg gctgctcagg acccaagctg cccaacccca gcactccccc 8220 aaagtgcctg ccaaaccctg ggcctggccc tgcccagtcc cactgcaagc cgtgtcctct 8280 ccctgactcc caagtgcctt tgcaccctag ggcccctaag tgcctgcccc cctccagagt 8340 attaacgctc caagagtatt attaatgctg ccgtacctca gtttgaacct gccagggccc 8400 cagctgctcc agcctgccag cagcttccag ccagtcccca cggccacgtc agctcaactc 8460 atcccttttt gatactatat cccctaccca gctacctatg gggcttgagg gttgtaaacc 8520 caaacaggtc agactccaat aaaggtgatt ctacagctgc 8560 <210> 17 <211> 2551 <212> DNA <213> Artificial Sequence <220> <223> Catalase <400> 17 gaagtcacca ctccagcggg cctggccaac aagattgcct tctccgggtg gagaccgctg 60 cgtccgtccc tgctgtctca cgttccgcag ctctgcagct ccgcaatcct acaccatgtc 120 ggacagtcgg gacccagcca gcgaccagat gaagcagtgg aaggagcagc gggcctcgca 180 gagacctgat gtcctgacca ccggaggcgg gaacccaata ggagataaac ttaatatcat 240 gaccgcgggg tcccgagggc ccctcctcgt tcaggatgtg gttttcactg acgagatggc 300 acactttgac agagagcgga ttcctgagag agtggtacac gcaaaaggag caggtgcttt 360 tggatacttt gaggtcaccc acgatatcac cagatactcc aaggcaaagg tgtttgagca 420 tattggaaag aggaccccta ttgccgttcg attctccaca gtcactggag agtcaggctc 480 agctgacaca gttcgtgacc ctcgggggtt tgcagtgaaa ttttacactg aagatggtaa 540 ctgggatctt gtgggaaaca acacccctat tttcttcatc agggatgcca tattgtttcc 600 atcctttatc catagccaga agagaaaccc acagactcac ctgaaggatc ctgacatggt 660 ctgggacttc tggagtcttc gtcccgagtc tctccatcag gtttctttct tgttcagtga 720 ccgagggatt cccgatggtc accggcacat gaatggctat ggatcacaca ccttcaagtt 780 ggttaatgca gatggagagg cagtctattg caagttccat tacaagaccg accagggcat 840 caaaaacttg cctgttggag aggcaggaag gcttgctcag gaagatccgg attatggcct 900 ccgagatctt ttcaatgcca tcgccaatgg caattacccg tcctggacgt tttacatcca 960 ggtcatgact tttaaggagg cagaaacttt cccatttaat ccatttgatc tgaccaaggt 1020 ttggcctcac aaggactacc ctcttatacc agttggcaaa ctggttttaa acaaaaatcc 1080 agttaattac tttgctgaag ttgaacagat ggcttttgac ccaagcaata tgccccctgg 1140 catcgagccc agccctgaca aaatgcttca gggccgcctt tttgcctacc cggacactca 1200 ccgccaccgc ctgggaccca actatctgca gatacctgtg aactgtccct accgcgctcg 1260 agtggccaac taccagcgtg atggccccat gtgcatgcat gacaaccagg gtggtgcccc 1320 caactattac cccaacagct tcagcgcacc agagcagcag cgctcagccc tggagcacag 1380 cgtccagtgc gctgtagatg tgaaacgctt caacagtgct aatgaagaca atgtcactca 1440 ggtgcggaca ttctacacaa aggtgttgaa cgaggaggag aggaaacgcc tgtgtgagaa 1500 cattgccggc cacctgaagg acgctcagct tttcattcag aagaaagcgg tcaagaattt 1560 cactgacgtc caccctgact atggggcccg catccaggct cttctggaca agtacaacgc 1620 tgagaagcct aagaacgcaa ttcacaccta cacgcaggcc ggctctcaca tggctgcgaa 1680 gggaaaagct aacctgtaac tccggtgctc agcctccgct gaggagacct ctcgtgaagc 1740 cgagcctgag gatcacctgt aatcaacgct ggatggattc tcccactccg gagcgcagac 1800 tcacgctgat gactttaaaa cgataatccg ggcttctaga gtgaatgata accatgcttt 1860 tgatgccgtt tcctgaaggg aaatgaaagg ttagggctta gcaatcattt aacagaaaca 1920 tggatctaat aggacttctg tttggattat tcatttaaat gactacattt aaaatgatta 1980 caagaaaggt gttctagcca gaaacatgac ttgattagac aagataaaaa tcttggcgag 2040 aatagtgtat tctcctatta cctcatggtc tggtatatat acaatacaac acacatacca 2100 cacacacaca cacatgcaat acacacacta cacacacata cacacactca cacacactca 2160 tacacacaca tgaagagatg ataaagatgg cccactcaga attttttttt ttatttttct 2220 aaggtcctta taagcaaaac catacttgca tcatgtcttc caaaagtaac tttagcactg 2280 ttgaaactta atgtttattc ctgtgctgtg cggtgctgtg ctgtgctgtg ctgtgcagct 2340 aatcagattc ttgttttttc ccacttggat tatgttgatg ttaatacgca gtgatttcac 2400 ataggatgat ttgtacttgc ttacattttt acaataaaat gatctacatg gaaggaccgt 2460 gtttggttgc tttcagctct gtataatgtg gaatgtgaag tagagattac cagctctctc 2520 tgcagtaaca ataaaagcgc cagcggccag a 2551 <210> 18 <211> 4124 <212> DNA <213> Artificial Sequence <220> <223> Signal transducing adapter molecule 2 <400> 18 agtcggagtc ccgccagagc gtcgggattc agctcgggtt gccggcgtga tgcctctgtt 60 cactgccaac ccgttcgagc aagatgtgga aaaagccaca aatgagtaca acaccacaga 120 agactggagt ctgatcatgg acatctgtga cagggttgga agcactccca gtggcaaaa 180 agattgccta aaagccataa tgaaaagggt aaaccataag gtccctcacg ttgctctgca 240 ggcattgacc cttcttggag cttgtgtggc aaactgtgga aagatatttc atttagaagt 300 atgttcccgt gattttgcaa cggaagtacg ttctgtgata aaaaataagg ctcatcctaa 360 agtatgtgaa aaactaaaat ctctaatggt ggaatggtca gaagaattcc agaaggaccc 420 ccaatttagt ctgatatctg caactattaa gtctatgaaa gaagaagggg ttacttttcc 480 ttccgcaggc tcccagactg tcgcggctgc tgccaaaaac gggacatcat tgaacaaaaa 540 caaagaagat gaggacatag ctaaagctat tgagttatcg ttgcaagagc agaagcagca 600 gtacacggag acaaaggctt tgtacccacc cgcagagagt cagctcaata acaaggctgc 660 acggagagtc agagctttat acgactttga agctgttgag gacaacgagc tcacctttaa 720 acatggtgag ctcattactg ttttggatga cagtgacgcc aactggtggc aaggagaaaa 780 tcacagagga acaggactct tcccctccaa ctttgtaacg actgacttaa gcacagaggt 840 tgagacagca acggtggaca aattgaatgt aattgatgat gatgtggagg aaattaagaa 900 atcagaacct gagcctgttt atatagatga gggtaaaatg gatagagccc tgcagattct 960 ccagagcata gatccaaaag agtcaaaacc cgactcccaa gacctcttgg acttggaaga 1020 tgtttgccaa cagatgggtc cgatgataga tgaaaaactt gaagagattg ataggaagca 1080 ttcagaactg tctgagttga acgtaaaggt gctggaagcc ctggacctgt acaataagtt 1140 ggtgaatgaa gcgcctgtgt actcggtcta ttcaaagctg catcccgcac attacccacc 1200 cgcagcagct ggggttccag tgcagacata tccagtccag tcgcatggtg gaaactacct 1260 gggccacggc attcaccaag tatctgttgc ccagaactat aacctaggac ctgatcctat 1320 gggctcgctg agatctctgc ctccaaatat gaactcggta acagcacaca ccgtccaacc 1380 tccatactta agcactggac aggacactgt ctccaaccct tcttacatga accagagctc 1440 tcgtcttcag gcagctgctg gtacagctgc ttacacacag cctgtgggga tgtctacaga 1500 tgtgtcttct ttccagaaca cagcatccgg tttgcctcag ctggctggct ttccagtggc 1560 agttcctgca cctgtcgctg cacagccaca agcaagctac caccagcagc ctctcctgta 1620 gagacgcacc aggacctgct gaacggcttt gtgggtgtgc tgtttaatct aggggattct 1680 aatctgaaat attaaaagtt tcccctctca gtcaaaaaga accatgaata aataaagcac 1740 aaaaccccac cttaccctga aggccataaa aggttttttt ttccagtcca gttggcttga 1800 agttgacctt ttggtcaaag gcagcttgga tgttctaagt tagtcgggct gaactttctg 1860 ggcccttact gagtagcctt acgacactac atgatatgag gtaggagtgt ggaattttct 1920 ctttaaagaa cccgttgttt cctcttaaga tgtgattttt gaggagcact gaagaccttg 1980 aaacattttt tgtctgcact tcccccaccc cttgaatttc ccacaccctg ttattttaaa 2040 aggcgagcat cttttagtgc aagcctttag cttgccagca tatttcctgt tggctcctta 2100 aattacagtt gtgtttgcag tactgtcagg tttgctctca gtgtaatgct gagtagtaat 2160 tagcagatac ttgtctacag aagcaagaac aatttggaag gtacaaaaat ggtttctgtc 2220 attaccagtg aaaaccatgt aaatgcaaat attgtggagc agttagccag tcctctgaca 2280 gacgagggcc agggactgcc ccaggagatg ttagccttcc ccagaaaggg catcctctca 2340 tactcgtgtc cctggtattt agaattgttt tctagtgtct tttggtggat gtctaatgaa 2400 ggtgctgtcc tttggatttc ctgcctgcta catatgcaga cactgcactt ccttcttagg 2460 gattctcagt gaatctgagc caggagacac tccccataac ctgatgtgat aggaaccatc 2520 atgggtatca gtgtgaggtg ggcgttcctt tcttacagaa agctgtcatc gtgtgccatt 2580 gggaagggat tgtgtgtccc ctagaagtag ttgtgagcct ttgcaggagc agcaggcagc 2640 ggtgtggtgg ccacaggctg acaggacagc agtcagctgc ttgtctgaag ctcctgtctt 2700 acgttaggtc ttagtacagt tccacatact gtaagagttg gtggcaaacc caagggaatt 2760 ttctcttaca gctttgcaca ctactttatt atatcagctt tttacacaac ataaactaga 2820 aaacatagat gcacaggact cggatccatg atatttacac tgggaaatgt tctaaatggg 2880 attttttaaa atttaaaatt tttgcatgaa tgcacttctg tatatttttt taagccctta 2940 aactttatta aaaatccatt gtgagacaat ttggaaatct tatcgtttat ctgtttacga 3000 tacttggtta taagtatgtt ttttttccaa agaaactgta ttctagaacc tactactgca 3060 taaaatagcc agcacagaag agagcgtggg ccgtccctgc aagcctgtat cttgtcagtg 3120 tctggttgtc agatgcagga ccctggaagg cttttcagtc cttagccctc accttctgtg 3180 tttcacacgg ccttctccct tcagctggac tcattgagtc cttacaagga tggagtgcgt 3240 gcaaaatggt accttactgt ttctagaaca gccttctgct cttcaaagat taagtaaata 3300 aatgtaaaag tccagagctg cccttgataa cgtttgtgta gaggaatgcg tcttccctgc 3360 ccttccaaac tgtgtttaat aagctcatgc tagtgtttgt gcatttcatg ttcagttatt 3420 gtctaaatta taaaagcttt atttagtcag tgtaaatact gcgttctcat taagtgggga 3480 ccagccattg aagcttagcc gttttggaat actaaaatat gccacatgac ctgtatataa 3540 atagtgtata ttagtgtctg tgaagtttaa tatatgtaga aatttcttcc tcctacattt 3600 tatttttttc cattcataaa tttgtgttgc agagttattg ttagcctaaa atattaaatc 3660 tgtattgatc tgatttcagt attaatacga tatgtaaaga tatatatttt cccatgatcc 3720 agttgaactg tacttatttt aagatgtttg tagctattta ttttcagcca aaggaatagt 3780 ttaccttcag atttctacat tgggttcata tttaatgttt tctaacaaaa atttttgaat 3840 ttcatttttc atataaatct tgatatatta gaaaattctt aaaaatatat agaattttat 3900 agtataaaac atttccttgg gtgttaagtg gagtatgttt gaaaagtcaa cagtcagcca 3960 atgttgtctt tttatttggt aatttgctta caaaattacc tacacattat ctttttgctt 4020 agcatttaaa tatgtatttc cattatgcta tttcaagtac ttgatttttg actaatgatt 4080 gtactgtata ctgaagatga taataaagtt tcatccacat tttg 4124 <210> 19 <211> 3705 <212> DNA <213> Artificial Sequence <220> <223> Beta-parvin <400> 19 ggccggcggt gggtggcgcg cactcgctta tgtcctccgc gccaccacgg tcgcccaccc 60 cgcgggcccc caagatgaag aaggacgagt ctttcttggg caagttgggc ggcacactgg 120 caaggaagaa gaagaccagg gaggtgactg acttacagga agaaggcaag agcgccatca 180 actctccaat ggccccggct ttggtggaca ttcaccctga agacacccag ctagaggaga 240 atgaggagcg taccatgatc gatcccacct ctagagaaga ccccaagttc aaggagttgg 300 tcaaggtact tctggactgg atcaacgatg tgctggcaga ggagcgcatc attgtgaagc 360 agctggagga ggacctgtac gatggccagg tgctccagaa acttctggaa aagctggcgc 420 actgcaagct gaatgtggca gaggtgacgc agtccgagat cgggcagaag cagaagttgc 480 agacggttct ggaagccgtg caagacctgc ttcggcccca tggctggccg ctgcgttgga 540 atgtggactc tatccatggg aagaacctag tggccattct gcatctcctc gtctccctgg 600 ccatgcactt cagggccccc attcaccttc ctgagcatgt caccgtgcag gtggtggtcg 660 ttcggaagcg agaaggcttg ctgcactcca gccacatctc agaggagctg accacgacca 720 cagagatcat gatgggccgg tttgagcggg acgccttcga tactcttttc gaccacgccc 780 cggacaagct caaccttgtg aagaagtctc tcatcacgtt tgtgaacaaa cacctgaata 840 agctaaactt ggaggtgact gacctggaga cccagtttgc agatggtgtg tacctagttc 900 tacttctggg ccttcttgaa gactactttg tcccccttca caacttctac ctgactcctg 960 acagctttga ccagaaggtg cacaacgtgg cctttgcctt tgaactgatg ctggatggag 1020 gcctcaagaa gcccaaggca cgccctgaag atgtggtgaa cttggacctc aagtccactc 1080 tgcgggtcct ttacacgctg ttcaccaagt acaaggatgt ggagtgacgg agcaactgat 1140 cctccaggga caagtgctag gccagaaaaa tgggtgtatc catccacaca gtccttgctt 1200 tcccagaaaa cacctccctg gggaggcctc aggcttccct ggcctctgct ccctgtcccc 1260 tgcctggttt ggttttaaca cccccccacc ccaccccctt ccgccgtgtg gttccctagc 1320 tggttaggtc attgagaact tggtgcttca aataccctct cccgtttaag tttaaggcct 1380 cacttcttct ttttctctgc ctcccctccc cctccccccc cccccccccc cccggagaaa 1440 gatgaaaaag agcaaaggcc cagcctagcc ctcggctgga gagggctgtc cacctgtgct 1500 ccagcccaca gagcaattac tgcaacagcc ctggctccag ggactccgcc ccatttgtcc 1560 tcaggaatcc cctgggctgg cctctccttg gaagcaaggg tctcagtaaa aaaagatcct 1620 ctcagttttt cactcctgag tctcagaaga tttagactcc tgatgtgtgt ctccacacac 1680 cccatcccca tccgtcttgc ggtttgctgc ttttaacgga ccttttacat gaactctcag 1740 gacccagaat gccttggggc cccggggtag tgtgtacggt gcatgtctgg gtctcttgct 1800 ggctctgaat gtgcagcttc agagccggct tggagtagga acctcctgtc tcggaggccc 1860 tgttagggaa tggacacatt tttagagact ttacaagaag gtagggatcc attacaaggg 1920 ctgcctgacc acaggctacc agcgagcagc cagaattaca gaggccgtgt gtctgactgc 1980 tctggaggct gagggtagaa ggtggaggtg ctggcaggct gcctttctct gccttcacac 2040 cattcctcct cctcctcctc ctcctcctcc tcctctattt atctcctctt cattccacag 2100 tggaaccatc ccctgcatga cctcgctgac cccatgatgt cagcaagcca cccctccctc 2160 cccccgcccc ccagtcgcca aatgcagcct ccctccgacc agccaggaat tggtttgtgg 2220 gctgtgtctc agctggcagt gggacctaag agatgtccac atccctccta gacactgctg 2280 gtatccccta cagcatgacc tataagagaa aatcctgctg ctccctcaag cccttcacca 2340 cccacttccc ttgactctgg aggtggtgcc atggccggct ggctccttgt ctcaggaatc 2400 ctgcttctgt ggttcacccc cacccccacc cccttctcac gttcctcctg cctctcacct 2460 gccttacagc actgttcatg gcagcttaca ggaaaccttc ctttcctgat tcccacctta 2520 ccacaagacc cagggctgtg gggtgaggtg ttgctaccga gctgaacgcc agcaatgatg 2580 ttccagaaaa cattttaata tcttcccttg gttccactgc tgctaagctg gggacgggtc 2640 ttggaatagc cgctccggtg gaggaggctt cccagcaggg gagagagata attaaaatgg 2700 cattaccgtg tctccctgtg ggatgcggtg acattaaaga gccacactga caaaataccc 2760 gggactggaa ggttctgtgc tgccttcctc gcagacacag aaccacagca gtatctggga 2820 gctgctggga ccgctttgct ctgctcacag gcggtctggg gcggggatcc tagatgcaaa 2880 gacctaccgt gctgaaggga gggaaagaat tggtctggga cgggcggtgg cttcctgggg 2940 ttccctatct ggagggcaca agtcctgctt ggaatgtaag caggctcctt tgcctgaggc 3000 agaacttggg aagccagggt tctatgaagg tgtgacctcc cccattgact gagcgtggag 3060 gtgtggttct cacacaccag tgaagggctg gccttaccgt cccttcactg cggagctgcc 3120 attcatacca tagcctggac aaaggcacca ataactaaga tgtctaggcc aacttcaggg 3180 ccacccgcgt aaggcaaggc tttcttcgta gaaaaaaaag agagaacccc cacccagatg 3240 acctcaggtg accacctgccc acccccaaac tttcccacag ttctctgggt ggggggctgg 3300 gatacaaata cttctctttg agctgttaat ttgagggcat tctggaacct tctactgctt 3360 ggatcttcat tctctcctga tattgttgag aggtctggcc actaaaactc ttagaaatgt 3420 catcatctgt cacttccatg gggtcttgag gaaggggcag gcaggtctgc accccctgcc 3480 cccaggtctg caggccagga ctgggttcct cccttctgcc ctgccctgat tggcacatcc 3540 cctccagctc tccagccttc tgtggacctg caggcagctc ttgtgccacc ttcacactca 3600 gttgtgcaag tcctgagctc aggccccctg catccttcct cattggctga ccaaggggaa 3660 tcttatatcc aagcagccga atccgttacg tcccctgtgc ggccg 3705 <210> 20 <211> 2177 <212> DNA <213> Artificial Sequence <220> <223> Guanine nucleotide-binding protein G (i) subunit alpha-2 <400> 20 gtcgctcgga actgcggacc tgagagcttc ccgcagaggg ccggcggtgg gagcggagtg 60 ggtcgggcgg ggccgagccg ggccgtgggc cgtgtggggg ccaggccggg ccggcggacg 120 gcaggatggg ctgcaccgtg agcgccgagg acaaggcggc agccgagcgc tctaaaatga 180 tcgacaagaa cctgcgggag gacggcgaga aggcggcgcg ggaggtgaag ttgcttctgt 240 taggtgctgg agagtcaggg aagagcacca tcgtcaagca gatgaagatc atccatgaag 300 atggctactc agaagaggag tgccggcagt accgtgccgt ggtctacagc aacaccatcc 360 agtctatcat ggccatcgtg aaggccatgg gcaacctgca gatcgacttt gctgatcccc 420 agcgtgcgga tgatgccagg cagctgttcg ccctgtcctg tgctgcagag gaacaaggga 480 tgcttcctga agacctgtcc ggtgtcatcc ggaggctctg ggctgaccac ggtgtgcaag 540 cctgctttgg ccgctcacga gaataccagc tcaatgactc agccgcttac tacctgaatg 600 atctggagcg cattgcacag agtgactaca tccctacaca gcaggatgtg ctgcggaccc 660 gtgtgaagac cacgggcatc gtggaaacac acttcacctt caaggactta cacttcaaga 720 tgtttgatgt gggtggtcag cggtctgagc gcaagaagtg gatccactgc tttgagggcg 780 tcacggccat catcttctgt gtcgccttga gcgcttatga cttggtgctg gctgaggatg 840 aggagatgaa ccgcatgcat gagagcatga agctgtttga cagcatctgc aacaacaagt 900 ggttcacaga cacctccatc atcctcttcc tcaacaagaa ggacctgttt gaagaaaaga 960 tcacacagag ctccctgacc atctgtttcc ctgagtacac gggggccaac aagtacgacg 1020 aggcagccag ctacatccag agcaagtttg aggatctaaa taagcgcaaa gacaccaagg 1080 agatctacac gcacttcacg tgcgccaccg acaccaagaa cgtgcagttt gtgttcgatg 1140 ccgtcactga cgtcatcatc aagaacaacc tgaaggactg tggcctcttc tgaggggcag 1200 tggacctggc aggatgggcc accgctgact gtgctcccca ctacccctga ggaagatggg 1260 ggcaagaaga ccatgttccc tgcctgttcc cccagctgct tctcccatct tttctctctg 1320 ttctcagctc ccctgtcccc tccctcggct ctagacttgg gggaggggtt gccacaggcc 1380 tcccagtcta aaacccacct ttgtctgagg tgccgggagt agccatggta cccccttcct 1440 gggcatccgt tcgggttttc taaccgttgt cttgttctgg ggtgagcggg gagcgcatgc 1500 agagatccca aggcctatgt ctggagggta ccaactcctc cagcctagac ccctggcttt 1560 gtccaacacc agccctgacc caagtccaaa tgtttacagg gagcctcctg cctaccccac 1620 tcctgccgc tcggaggccc caaaggaaaa agcacaagaa gcgtgagaga taccgccatt 1680 cctggagaca aagcccacct gctcattctc gtagctttaa agaaaaagaa aaaggaaaaa 1740 taaggaaaac tgcaaatcta gaaaactttt tagagaaacc tatttaaaac tgtcaggtcc 1800 tgaccagcac ccccacccca gcccagcccc agccccaccc ctttccagcg attccgtgcc 1860 ttgagtgtgt ctgcgtgttt acacccatcc ctctttgggc ggccccctgc tctgccctcc 1920 acggaattgg attccaaggg ctgttccaga caactgccaa cgtcactgag ggccctgctc 1980 tgaagccctg ggccctggct ccattaacct aaatgtagct ccttagcgct aatctaggaa 2040 ccgccgctgc ctgctgaggg ccaggcccct cacgccctcg ccccaggccc gggtcctcca 2100 gcgttgaaca cttccttgct tttttcacat gttttatgga attgttcaca tgatttgaaa 2160 taataaaatg tagaaag 2177 <210> 21 <211> 4447 <212> DNA <213> Artificial Sequence <220> <223> Delta-aminolevulinic acid dehydratase (porphobilinogen synthase) <400> 21 gagcgcggtg tccagagccc ggctcggagc ggcggcgagc agcgtccttg gtacgtgagc 60 aggcgccggc ctctgggaag tggcgggttg aagggggacg ggaccccaga acaagagctg 120 aggctgcagc agacacccag gagccgccac tccatccagc agacttctct gtgttccact 180 gccccgacca tgcaccacca gtctgttctg cacagcggct actttcaccc actgcttcgg 240 agctggcaga ctgctgcctc caccgtcagt gcctccaacc tcatctatcc catctttgtc 300 acggatgttc ctgatgatgt ccagcctatc gccagcctcc caggagtggc caggtatggc 360 gtaaaccagc tagaagagat gctgagacct ctggtggaag ctggcctgcg ctgtgtcctg 420 atctttggcg tccccagcag agttcccaag gatgaacagg gctctgcagc tgactctgag 480 gactccccaa ctattgaggc tgtccgtctg ctgaggaaga ccttcccttc cctcctagtg 540 gcctgtgacg tctgcttgtg cccctacacc tcccatggcc actgtggcct cctgagtgaa 600 aatggagcat tcctagcaga ggagagccga cagcggttgg cagaggtggc actggcctat 660 gccaaggcag gctgtcaggt tgtagctccg tcagacatga tggacggacg agttgaggcc 720 atcaaggctg ccctgctaaa acatggactt ggcaacaggg tctctgtgat gagctatagt 780 gccaaatttg cctcctgttt ctacggtcct ttccgggatg cagctcagtc aagcccagct 840 tttggagacc gacgctgtta tcagctgcct cctggtgccc gtggcctggc cctccgagca 900 gtggcccgag acattcaaga aggagctgac atgctcatgg tgaagccggg attgccctac 960 ctggacatgg tgcgagaggt gaaggacaag caccccgagc tccccctcgc agtataccag 1020 gtgtctggag agtttgccat gttgtggcac ggagcccagg ccggggcctt tgatctcagg 1080 actgctgtac tggagaccat gacggccttc cgcagagccg gtgccgacat catcatcacc 1140 tactttgcac cgcagctgtt gaagtggctg aaggaagaat gaagaaaaag tgtcggactt 1200 gatcttgacc aagctccctg ggccttacag aagggggaaa gtaaatgcgc tgttagaact 1260 gtgccctgtg ccctcttcct gcccacgctg ctggcagggt gcagcgccct gggtggtttg 1320 ccagcatgct aaactcccac tcacagctgc agcctaccag gccctccaag tttcccactc 1380 tggctcagcc cagagaccct cctccctgct ctgctgaggt gtggccttag cttggcagaa 1440 cccaagagcc tcaggcaccg gcttggagct ttgggacaag aggagagcag ttggagcttg 1500 aaacctaaag agttattcat taatgaggat caaaccccag aagcagcagg gattcctggc 1560 cacaggagat gccagcatca gaaagtctgt ctgcacaagt cagttcctga gaacctctgc 1620 ccagggcaga tggacacagc aggatgcaca gcagcatacg tctctgtagg atgtatgcag 1680 agaaagggga gcgtaacaaa gcaggtgaat cgatacttgc taagaagggg aagtccaaga 1740 cagctgtccc tgagttgaaa ccttaaaggg acaaagagga ccagcagacg tgctcagaga 1800 cccctttggt aggaggagct tggcagggct agagggcagg cgacgaagag gaacgtggat 1860 aacagagggg aggttggaat agttcgttcc ccgacttagc agggacctaa agttttgttt 1920 tgcatctgtg ggacagccat cttaaagatt tgggcgtttg gtttagtttt ctccactaag 1980 tatccctggc ctcacccacc taggcaagca ctctgccact gaactgtacc tatagccttc 2040 aaggcacgtt ctggagacca gactagatgt gttgcgggag gtagataagg gaacaagaga 2100 gagtgaggtt gtctttgtgg gtttctgaag ttgcccagct ggttttagtt cattcatctg 2160 acatgacacc cgtactgcgg ctaatgaagg ttagggccag tttgcaggca cacctgactt 2220 tagaggtgga ggccctactc tagctctgac tctaacgcga tgtcattctt aaggtgcaga 2280 attagttaga gttaataact ggcaactcct gcaatcagac tggtccaata cttggtggct 2340 ttcttttaaa gctgtgaata actcaagagc tgatcttcct acttctcagc cttctggaag 2400 gtagccacca aaatcaaatc agccctaaac tgtctctttc ttggatcctg agcatagatt 2460 taccatctag aacccagatc actaatttaa aacctgctat catcaggcca ggggtatacc 2520 agactagact tgtactatct tggctgttat acagcctctg agctctgtgg ttaaaatggt 2580 tcgtgtgtta gggaagccat caccaagctg ttaagagagc caggagacat gctaccctgc 2640 atgtgccaag gccctaatgt gtccaggccc aatggcaaat gggaaggtca ccaggtcccg 2700 acctttgaag gatcagaaaa cagtcccctg aaggactgat gtttggatga acagctaaag 2760 ctgagacata attgaccaag agccttccag ggaaagaacc attgtgcaaa gttcctggag 2820 ccgaagtgga gctggatgag cacggggctg gtgtggcccg cacaccagga agaatacagg 2880 aacaggtggc ctggacatct cagtctaagc cgtacagggg cttcgatgcc acactcctcc 2940 acctttatcc caagatcaaa gggcacctca aggatgccag caggggaatg accatgttgt 3000 gggttttcac ttagtaacac catggcatta tgcagatgag atgtttgaat tccatggtga 3060 ccatagggat gcaaaaaagg agattattta gagtatagtt tagacctgct tctctgaatg 3120 gcaagtggcc tgaccattta atggagaaag cagactcaag gtagaaccct aagtttagct 3180 tgagtccttg gcaatgagaa aggcaccccc cactaaggct actcacattt tcggcagaac 3240 tgggttaaag gttaaaagga aaaccagatg taacattctg acccagctcc aaagccccga 3300 cccttaacca ctcatgacac ctcccagtaa gggacagtga gaaagatggc tggcctctgc 3360 tgtgtgacta actggtatgg gaggaggacc ttggagccca ttttcccctg gagctgggtc 3420 ttgtcacctg aagtgtccac ttggtggcag ccacctacat ctttgcaaac aaagcaggca 3480 cagacagaca tggcttacac ctaagggctt cctgcacaga agggcttcct ggacttcaag 3540 actaacctaa gctgcagaat aagatccttc tttaaaatcg ggaagagact tgagagtcat 3600 ctgaaagggt tgtaactgac ctgtgccaca tccatagaga actagatttt cagtctggca 3660 aggatattat cagcctccca gatacctggc agtgggtgtc cagggacttg atgagacttg 3720 gttttctcat tcccctctca aatgtgccag tttaggctgt agtttcaccc tttggcaggc 3780 atcataatca gccatttaac tccttaagag tggagaacac ccagtttcca gctagccatc 3840 ccatatagac tgccagtcta gcacaggaag ggcctagtga tttgtggcag tcttgagtct 3900 gggtactctt ggttgcaggg actgaaactc aggtcaatag aaaaagaaac tgaactggtt 3960 cgcatgccta aatcttggta atgtcaatta tgtgtgtccc caagtcaaag gctttgatct 4020 tttggccttg gggagacttt attgttggat aagccttccc agccaacaac ttgatccctc 4080 actgaaataa cccctaagat ttgtaatccc agagaaagac ccattctgtc ctgtccctgc 4140 cataacaact ctggagatct gtgattgttc tgcgtaggtc acatgtctgc caattggctc 4200 actcagtatg aactggacag ggatgggtga catggttgac ctgccccccc attacataaa 4260 aggctgacaa ggccttggct ctggtaccaa atggtcctct gcctggaatg ccttctctct 4320 ctcagctgc atccagcaat ctctctaaac tcaccctctg tattcggatt agtgaataca acaagtcaaa 4440 appointment 4447 <210> 22 <211> 1854 <212> DNA <213> Artificial Sequence <220> <223> L-lactate dehydrogenase <400> 22 gggtttggg gggggtgggg gggttaggaa ggaagcttgc gcgtgcgcag gcttaagcac 60 gttgctatgc cttggggtcg caccttgtgg ccgttattgg cgccctctgc tcttgatttt 120 tggtacttcc tggagcaact tggcgctcta cttgctgtag ggctctgggt gatgggagaa 180 gagcgggagg gcagctttct aaccatataa gaggagatac catccccttt tggggttcat 240 caagatgagt aagtcctcag gcggctacac gtacacggag acctcggtat tatttttcca 300 tttcaaggtc tcaaaagatt caaagtccaa gatggcaacc ctcaaggacc agctgattgt 360 gaatcttctt aaggaagagc aggctcccca gaacaagatt acagttgttg gggttggtgc 420 tgttggcatg gcttgtgcca tcagtatctt aatgaaggac ttggcggatg agcttgccct 480 tgttgacgtc atggaagaca aactcaaggg cgagatgatg gatctccagc atggcagcct 540 cttccttaaa acaccaaaaa ttgtctccag caaagactac tgtgtaactg cgaactccaa 600 gctggtcatt atcaccgcgg gggcccgtca gcaagagggg gagagccggc tcaacctggt 660 ccagcgaaac gtgaacatct tcaagttcat cattcccaac attgtcaagt acagtccaca 720 ctgcaagctg ctgatcgtct ccaatccagt ggatatcttg acctacgtgg cttggaaaat 780 cagtggcttt cccaaaaacc gagtaattgg aagtggttgc aatctggatt cagcgcggtt 840 ccgttacctg atgggagaga ggctgggggt tcacgcgctg agctgtcacg gctgggtcct 900 gggagaacat ggcgactcca gtgtgcctgt gtggagtggt gtgaatgttg ccggcgtctc 960 cctgaagtct cttaacccag aactgggcac tgacgcagac aaggagcagt ggaaggaggt 1020 tcacaagcag gtggtggaca gtgcctacga ggtgatcaag ctgaaaggtt acacatcctg 1080 ggccattggc ctctctgtgg cagacttggc tgagagcata atgaagaacc ttaggcgggt 1140 gcatcccatt tccaccatga ttaagggtct ctatggaatc aatgaggatg tcttcctcag 1200 tgtcccatgt atcctgggac aaaatggaat ctcggatgtt gtgaaggtga cactgactcc 1260 tgaggaagag gcccgcctga agaagagcgc agacaccctc tggggaatcc agaaggagct 1320 gcagttctaa agtcttcccc gtgtcctagc acttcactgt ccaggctgca gcagggcttc 1380 taggcagacc acacccttct cgtctgagct gtggttagta cagtggtgtt gagatggtgt 1440 ggggaaacat ctcactcccc acagctctgc cctgctgcca agtggtactt gtgtagtggt 1500 gacctggtta gtgtgacagt cccactgtct ctgagacaca ctgccaactg caggcttcga 1560 ttacccctgt gagcctgctg cattgctgcc ctgcaccaaa catgcctagg ccgacgagtt 1620 cccagttaag tcgtataacc tggctccagt gtgtacgtcc atgatgcata tcttgtgcat 1680 aaatgttgta caggatattt tatatattat atgtgtctgt agtgtgcatt gcaatattat 1740 gtgagatgta agatctgcat atggatgatg gaaccaacca cccaagtgtc atgccaaata 1800 aaaccttgaa cagtgaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaa 1854 <210> 23 <211> 1966 <212> DNA <213> Artificial Sequence <220> <223> T-complex protein 1 subunit delta <400> 23 ggctctccag caggcccctt gctcggcttc cgccttctct cccggcgtcg ctttctggaa 60 ggttcgtgga ggaggcggtc agggagaccg ttactccaca gcaagccgga atccgtgtcc 120 atccgtcctc ctgaacccgc gcagacgcca ccaaggtcgc catgccggag aacgtagctt 180 cccgaagcgg ggcgcccacc gccgggcccg gcagccgcgg gaaaagcgcc taccaggacc 240 gcgacaagcc agcccagatc cgcttcagca atatttccgc ggccaaagcg gttgctgatg 300 ctattagaac aagccttgga cctaaaggaa tggacaaaat gattcaagat ggaaaaggcg 360 atgtgaccat tacaaatgat ggtgccacca ttctgaaaca aatgcaggta ttgcatccag 420 cagccagaat gctggtggaa ttgtctaaag ctcaagacat agaagcagga gatggcacca 480 cgtcggttgt catcattgct ggctctcttt tagactcctg taccaaactt ctgcagaaag 540 gtatacatcc aaccatcatt tccgagtcat tccagaaagc tttggaaaag ggtcttgaaa 600 tccttactga catgtctcga cctgtgcaac tgagcgacag agaaactttg ttaaatagcg 660 caactacttc attgaattca aaggttgtct ctcagtattc aagtctactc tctccaatga 720 gtgtcaatgc ggtgatgaaa gtgattgacc cagccacagc taccagtgta gatcttcgag 780 atattaaaat agttaagaag cttggtggga caatagatga ctgtgagctg gtggaaggcc 840 tcgttctcac acagaaagta gcaaattctg gcataacaag agttgaaaag gctaagattg 900 ggcttattca gttttgctta tctgctccta aaacagatat ggataatcaa atagtagtat 960 ctgactatgc ccagatggat cgagtgcttc gagaggagag agcctatatt ttaaatttgg 1020 tgaagcaaat taagaaaaca ggatgtaatg tccttctcat acagaagtct atcctgagag 1080 atgcccttag tgatcttgca ttacattttc tgaataagat gaagattatg gtggttaagg 1140 acgttgaaag agaagacatt gaattcatct gtaagacaat tggaaccaaa ccagttgctc 1200 acattgacca gttcactgct gacatgctgg gttctgctga gttagcagag gaagtcagtt 1260 taaatggttc tggaaaacta ttcaagatta caggttgtac aagcccaggg aaaacagtta 1320 caattgtcgt acgtggttct aacaaactgg tgattgaaga agctgagcgc tccattcatg 1380 atgctctctg tgtcatccga tgcttagtaa agaaaagagc tcttattgca ggaggtggtg 1440 ctccagaaat agagctggcc ctcagactga cagagtactc ccgaacactg agtggtatgg 1500 agtcctactg tgttcgtgct ttcgcggatg ctatggaagt cattccatct acactagctg 1560 aaaatgctgg cctgaatccc atttctacag taacagagct aagaaaccgc catgcccaag 1620 gagaaaaaac tacaggcatt aatgtccgaa agggtgggat ctccaacatt ttggaggaaa 1680 tggttgttca gcctctgttg gtgtcagtca gtgctttgac cttagcaact gaaactgtgc 1740 ggagcattct gaaaatcgat gatgtggtaa atactcgata atctggataa aaggatggtt 1800 gactgcatca ttatggacag aagtactgtg gctggaatga aggacaacca ccttgttcct 1860 tgtctggaag cttcagagtt tttggacatt gtcttccagt tggcatttgt ttgaaattgt 1920 attgaaacaa tttaatgaaa acattaaata cttggtttta aactcc 1966 <210> 24 <211> 1446 <212> DNA <213> Artificial Sequence <220> <223> peroxiredoxin 2 <400> 24 gcccacctcc ccgcgcactg cctggtgtgg tttgggccac gcataaaagg ttctccggcc 60 tagggctctc tcggttttga gatctctttc ctgtctctac ccgtgtctgg acgtccgtgt 120 gtccggctct tgctcacgca gacttagcac caaaagtgac cttggaaagg caagacaaag 180 gatcttctaa ggctctaaag accaagcttg tgtgggtctc tgatttcagt catggcctcc 240 ggcaacgcgc aaatcggaaa gtcggctcct gacttcacgg ccacagcggt ggtggatggt 300 gccttcaagg aaatcaagct ttcggactac agagggaagt acgtggtcct ctttttctac 360 ccactggact tcacttttgt ttgccccacg gagatcatcg cttttagcga ccatgctgag 420 gacttccgaa agctaggctg cgaggtgctg ggagtgtctg tggactctca gttcacccac 480 ctggcgtgga tcaatacccc acggaaggag ggaggcttgg gccccctgaa tatccctctg 540 cttgctgacg tgactaaaag cttgtcccag aattacggcg tgttgaaaaa tgatgagggc 600 attgcttaca ggggtctctt tatcatcgat gccaagggtg tccttcgcca gatcacagtc 660 aatgacctac ctgtgggacg ctctgtagac gaggctctcc gcctagtcca ggcctttcag 720 tatcatacg agcatgggga agtctgccct gctggctgga agcccggcag tgacaccatc 780 aagcccaatg tggatgacag caaggaatac ttctccaaac acaactgaga tgggtaaaca 840 tgggtgagcc tgaagcttgg atttcacctg tgccccaacc tggatgtcct gtgctggccc 900 agaaaatgct agattttcct ccactctctg aaggggctgg agtctaggct gaggttttct 960 cattacccac ctggaatctg gtgaatagtg atcctgccct gagcacacct agctgggccc 1020 aggtctatag gaaaccaata aagtattagg gacagtgtaa gtctgtgttg gtgtgtgttt 1080 agtagtttta gtagtttaca ctctggcttg tgcaactgac tctgtagtta tgcctcttca 1140 ccacaaggtg gaggcattac ctcattcacg ccatggtctg agcatggtgg gacacagtga 1200 gccaaataa ttactaggcc tggagggatt ctttaagagc tctttatgtt tctcctgaag 1260 atccaggttc cattcctgga gcttacatag ttgcttacaa ccctctttaa cgccagatcc 1320 aacaccttcg tctgatcctc cagcaccagg catgcacgtg aacatgggca gaacgtgcat 1380 agacatgaaa taaatacttc tgtaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 1440 aaaaaa 1446

Claims (16)

a) Heat shock cognate 71 kDa protein, ITIH2 (inter-alpha trypsin inhibitor, heavy chain 2), DBP (vitamin D-binding protein), and TLN1 from blood samples isolated from subjects requiring diagnosis of normal persons and Alzheimer's disease (TA), CAT (catalase), STAM2 (signal transducing adapter molecule 2), PARVB (beta-parvin), GNAI2 (guanine nucleotide-binding protein G (i) subunit alpha-2), ALAD (delta-aminolevulinic acid dehydratase, LDHA (L-lactate dehydrogenase), CCT4 (T-complex protein 1 subunit delta), and PRDX2 (peroxiredoxin 2), and
b) the subject is judged to be an Alzheimer's patient if the amount of protein detected in step (a) is increased or decreased in the subject compared to a normal subject,
How to diagnose Alzheimer's disease. 2. The method of claim 1, wherein the protein having an increased amount in the subject as compared to a normal person is selected from the group consisting of HSPA8, ITIH2, DBP, TLN1, CAT, STAM2, PARVB, GNAI2, ALAD, LDHA, and CCT4 Gt; Alzheimer &apos; s disease. &Lt; / RTI &gt; 2. The method of claim 1, wherein the protein having a reduced amount in the subject as compared to a normal subject is PRDX2. 4. A method for diagnosing Alzheimer's disease according to any one of claims 1 to 3, wherein the detection of the protein detects an antigen-antibody complex formed by the protein or an immunogenic fragment thereof and an antibody thereto . The method according to claim 4, wherein the detection of the antigen-antibody complex is carried out by Western blotting, enzyme linked immunosorbent assay (ELISA), immunoprecipitation assay, complement fixation assay, fuorecence activated cell sorter (FACS), or protein chip. The present invention relates to an exosomal protein which is increased or decreased in the blood of a patient suffering from Alzheimer's disease as compared with blood of a normal person. The exosome protein is selected from the group consisting of HSPA8, ITIH2, DBP, TLN1, CAT, STAM2, PARVB, GNAI2, ALAD, LDHA and CCT4 A composition for diagnosing Alzheimer's disease, comprising as an active ingredient an antibody that specifically binds to one or more selected proteins or immunogenic fragments thereof. 7. The composition of claim 6, wherein the antibody is a monoclonal antibody, a polyvalent antibody, or a recombinant antibody. The present invention relates to an exo-somatic protein which is expressed by HSPA8, ITIH2, DBP, TLN1, CAT, STAM2, PARVB, GNAI2, ALAD, LDHA, CCT4 and PRDX2 in a relatively increased or decreased expression in the blood of a patient with Alzheimer's disease 0.0 &gt; 1, &lt; / RTI &gt; wherein the expression level of at least one protein selected from the group is selected from the group consisting of: 9. The kit for diagnosing Alzheimer's disease according to claim 8, wherein the agent for measuring the protein expression level is an antibody that specifically binds to the protein or an immunogenic fragment thereof. a) preparing a blood extracellular vesicle from a blood sample separated from a subject;
b) a gene selected from among the genes of Hspa8 , Iyih2 , Gc , Tln1 , Cat, Stam2 , Parvb , Gnai2 , Alad , Ldha , Cct4 and Prdx2 , which are constituted by the nucleotide sequence selected from the group consisting of SEQ ID NOS: 13 to 23 and SEQ ID NO: Measuring one or more levels of expression; And
c) a method of providing information for monitoring or diagnosis of Alzheimer's disease, comprising determining that one or more of the genes of b) is increased or decreased compared to a normal control, . 11. The method according to claim 10, wherein the expression level of b) is one selected from the group consisting of RT-PCR, ELISA, Western blotting, and immunohistochemistry, and provides information for monitoring or diagnosis of Alzheimer's disease How to. a) measuring the expression level of at least one exosome protein selected from the group consisting of HSPA8, ITIH2, GC, TLN1, CAT, STAM2, PARVB, GNAI2, ALAD, LDHA, CCT4 and PRDX2 from a biological sample isolated from the subject; And
b) determining if the subject is at increased risk of Alzheimer &apos; s disease or is an Alzheimer &apos; s disease subject if the level of expression of the protein of step a) is increased or decreased compared to a normal control, A method for measuring the level of exosome protein expression to provide diagnostic information. 13. The method according to claim 12, wherein the biological sample of step a) is selected from the group consisting of bone marrow, serum, plasma and blood. 14. The method according to claim 12 or 13, wherein the measurement of protein expression level in step a) uses an antibody that specifically binds to each of the proteins. a) treating the test substance with an extracellular fibrin cell isolated from a patient suffering from Alzheimer's disease;
b) measuring the expression level of at least one exosomal protein selected from the group consisting of HSPA8, ITIH2, GC, TLN1, CAT, STAM2, PARVB, GNAI2, ALAD, LDHA, CCT4 and PRDX2 in the extracellular follicle of step a) step; And
c) selecting the test substance which increases or decreases the protein expression level of the step b) compared to the untreated control,
A method for screening candidates for treating Alzheimer's disease. 16. The method of claim 15, wherein the protein expression level measurement in step b) is selected from the group consisting of immunofluorescence, mass spectrometry, protein chip, Western blotting and ELISA.

Images