KR102146392B1 - Cell-penetrating phosphoglycerate mutase 1 fusion protein and pharmaceutical Composition containing thereof - Google Patents

Abstract

An object of the present invention is to provide a phosphoglycerate mutase 1 fusion protein and a pharmaceutical composition comprising the same, which may more efficiently transport phosphoglycerate mutase 1 into cells or tissues, thereby preventing or treating symptoms or diseases such as central nervous system diseases, muscle pains, convulsions, and muscle necrosis, etc., which are symptoms caused by phosphoglycerate mutase 1 deficiency. To solve the above problem, the present inventors selected and tested a number of candidate peptides, and thus found that a peptide consisting of 18 amino acids derived from human adenosine receptor A2a (ADORA2A) and a modified sequence in which several sequences are substituted, added or deleted by using the peptide as a base sequence bind to the mutase 1 protein to smoothly permeate this protein into cells and tissues. The fusion protein, and an oligonucleotide or a vector encoding the same can be applied as a pharmaceutical composition for preventing or treating a phosphoglycerate mutase 1 deficiency disease.

Description

Phosphoglycerate mutase 1 fusion protein and pharmaceutical composition containing the same {Cell-penetrating phosphoglycerate mutase 1 fusion protein and pharmaceutical composition containing thereof}

The present invention relates to a phosphoglycerate mutase 1 fusion protein and a pharmaceutical composition comprising the same, wherein the phosphoglycerate mutase 1 is fused to a new cell penetrating peptide to prepare a fusion protein having high cell penetration efficiency, and this , Convulsions, muscle necrosis or myoglobinuria prophylaxis or treatment relates to a method of manufacturing and applying a pharmaceutical composition for use.

Protein delivery technology is called Protein Transduction Domain (PTD) or Cell Penetrating Peptide (CPP). A peptide consisting of 5 to 30 amino acids is fused with a polymer such as a protein or gene to form mammalian cells and tissues. It is a new concept of delivery system that can be easily delivered into the living body such as blood and blood. Although the specific mechanism has not yet been accurately elucidated, protein delivery technology has been widely used to deliver therapeutic proteins into cells or tissues in vitro and in vivo, and a wide variety of protein transport domains are known.

Among the numerous protein transport domains, widely known is a cell-permeable HIV-Tat peptide, whose sequence includes RKKRRQRRR, and has been used in the study of various therapeutic proteins. In addition, it was found that the Pep-1 peptide can be used to transport heterologous proteins in a natural state into cells. Pep-1 peptide consists of 21 amino acids (KETWWETWWTEW SQP KKKRKV) and has three domains, namely, a hydrophobic domain, a spacer, and a hydrophilic domain. Until now, studies using the Pep-1 peptide have shown that when a foreign protein is covalently fused to the Pep-1 peptide and administered to a cell, the protein can be transported in a natural state into the cell.

Other widely known protein transport domains include oligolysine, oligoarginine, and oligo (lysine + arginine).

Phosphoglycerate mutase 1 (phosphoglycerate mutase 1; hereinafter used together with "PGAM1") is an enzyme that catalyzes the glycolysis process, and converts 3-phosphoglycerate into 2-phosphoglycerate. Patients with phosphoglycerate mutase 1 deficiency may experience symptoms of the central nervous system such as mental retardation and seizures. In addition, when phosphoglycerate mutase 1 is deficient, symptoms such as muscle pain, cramps, muscle necrosis, myoglobinuria, exercise intolerance, and proximal muscle weakness may occur.

It is an object of the present invention to more efficiently transport phosphoglycerate mutase 1 into cells or tissues, and symptoms or diseases such as central nervous system diseases, muscle pain, convulsions, and muscle necrosis, which are symptoms caused by a deficiency of phosphoglycerate mutase 1 It is intended to provide a phosphoglycerate mutase 1 fusion protein capable of preventing or treating, and a pharmaceutical composition comprising the same.

In order to solve the above problem, the present inventors selected and tested a number of candidate peptides as cell-penetrating peptides. As a result, the present inventors determined a peptide consisting of 18 amino acids derived from human adenosine receptor A2a (ADORA2A) (hereinafter, referred to as "GK2" in the present invention. Name) and a modified sequence in which several sequences have been substituted, added or deleted using this as the base sequence, reveals that polymers such as proteins can smoothly permeate into cells, tissues, blood, etc. Poglycerate mutase 1 was fused to produce a cell-penetrating phosphoglycerate mutase 1 fusion protein. Cell-penetrating phosphoglycerate mutase 1 fusion protein smoothly permeated into cells and showed activity in cells.

The present invention

(A) R ₁ R ₂ R ₃ RIREFRQTFRKR ₄ R ₅ R ₆ R ₇ ,

(However, R ₁ is _{one selected} from Y, R, RR, RRR, RRRR, RY, RRY, RRRY, R ₂ is one selected from A and R, R ₃ is one selected from Y and R, and R ₄ is one selected from I and R, R ₅ is one selected from I and R, R ₆ is R, R ₇ is one selected from S, R, RR, RRR, RRRR, SR, SRR, SRRR),

(B) R ₈ REFRQTFRKR ₉ ,

(However, R ₈ is one selected from RKKRRQRRRYAYRI, RKKRRQRRRI, RKKRRQRR and YAYRI, R ₉ is one selected from IIRS, IIRKKRRQRRR and KRRQRRR),

(C) R ₁₀ RQTFRKIIRS

(However, R ₁₀ is 19 or more consecutive amino acids in the KETWWETWWTEWSQPKKKRKV sequence),

(D) YAYRIREFR ₁₀

(However, R ₁₀ is 19 or more consecutive amino acids in the KETWWETWWTEWSQPKKKRKV sequence),

Cell-penetrating phosphoglycerate combined with a cell-penetrating peptide that is one selected from the above (a) to (d), consisting of 16 to 31 amino acids, and that binds to a cargo molecule to transport cargo molecules into mammalian cells or tissues It relates to a mutase 1 fusion protein.

In addition, the present invention is characterized in that the cell-penetrating phosphoglycerate mutase 1 fusion protein is one selected from SEQ ID NO: 49 to SEQ ID NO: 71. However, these amino acid sequences are only exemplary, and it is obvious to those of ordinary skill in the art that the type of cell-penetrating phosphoglycerate mutase 1 fusion protein is not limited thereto.

In addition, the present invention is characterized in that the bond between the cell-penetrating peptide and the phosphoglycerate mutase 1 fusion protein is a covalent bond.

In addition, the present invention relates to a pharmaceutical composition for preventing or treating a phosphoglycerate mutase 1 deficiency disease comprising the cell-penetrating phosphoglycerate mutase 1 fusion protein.

Phosphoglycerate mutase 1 deficiency diseases or symptoms include central nervous system disorders such as mental retardation and seizures, and muscle pain, convulsions, muscle necrosis, myoglobinuria, exercise intolerance, and proximal muscle weakness. Examples can be cited.

In addition, the present invention relates to a recombinant polynucleotide encoding the cell-penetrating phosphoglycerate mutase 1 fusion protein.

In addition, the present invention is characterized in that the recombinant polynucleotide is one selected from SEQ ID NOs: 72 to 94. However, these polynucleotide sequences are exemplary only, and it is obvious to those of ordinary skill in the art that the type of recombinant polynucleotide encoding the cell-penetrating phosphoglycerate mutase 1 fusion protein is not limited thereto. Do.

In addition, the present invention relates to a pharmaceutical composition for preventing or treating a phosphoglycerate mutase 1 deficiency disease comprising a recombinant polynucleotide encoding the cell-penetrating phosphoglycerate mutase 1 fusion protein.

In addition, the present invention relates to a fusion protein expression vector comprising a recombinant polynucleotide encoding a cell-penetrating phosphoglycerate mutase 1 fusion protein.

In addition, the present invention relates to a pharmaceutical composition for preventing or treating a phosphoglycerate mutase 1 deficiency disease comprising the fusion protein expression vector.

A pharmaceutical composition containing the cell-penetrating phosphoglycerate mutase 1 fusion protein of the present invention, an oligonucleotide encoding the same, or a vector containing the oligonucleotide as an active ingredient, is formulated with a carrier commonly accepted in the pharmaceutical field. It can be formulated in various forms such as skin external preparation, oral, spray, patch, or injection by a conventional method. For example, oral compositions include tablets and gelatin capsules, in addition to the active ingredient, diluents (e.g. lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and/or glycine), lubricants (e.g. silica, talc). , Stearic acid and its magnesium or calcium salt and/or polyethylene glycol), and the tablet also contains a binder (e.g. magnesium aluminum silicate, starch paste, gelatin, methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidone). ), and in some cases, a disintegrant (eg starch, agar, alginic acid or sodium salt thereof) or a boiling mixture and/or an absorbent, colorant, flavoring and sweetening agent. The composition for injection is preferably an isotonic aqueous solution or suspension, and the compositions mentioned are sterilized and/or contain adjuvants (e.g., preservatives, stabilizers, wetting or emulsifying agent solution accelerators, salts/or buffers for controlling the osmotic pressure). In addition, they may contain other therapeutically useful substances.

The pharmaceutical preparation prepared as described above may be administered orally, or parenterally, ie, intravenously, subcutaneously, intraperitoneally, or topically, as desired. The dose can be administered by dividing the daily dose of 0.0001 to 100 mg/kg in 1 to several times. The dosage level for a specific patient may vary depending on the patient's weight, age, sex, health status, administration time, administration method, excretion rate, and severity of disease.

The cell-penetrating phosphoglycerate mutase 1 fusion protein fused with the novel cell-penetrating peptide of the present invention can easily penetrate into cells.

In addition, since the new cell-penetrating phosphoglycerate mutase 1 fusion protein of the present invention exhibits cytoprotective activity in cells, a fusion protein, an oligonucleotide encoding it, or a vector containing the oligonucleotide is used as a phosphoglycerate mute. It can be used as a pharmaceutical composition for the prevention or treatment of Izu 1 deficiency diseases or symptoms.

Figure 1 shows the construction of the GK2 expression vector on the pET-15b vector. The synthetic GK2 oligomer was cloned into the Nde I site, and the human PGAM1 (Phosphoglycerate mutase 1) cDNA was cloned into the Xho I and BamH I sites of pET-15b.
Figure 2 shows the construction of the GK2 expression vector on the pET-15b vector. The synthetic GK2 oligomer was cloned into the Nde I site, and the human CNRIP1 (CB1 cannabinoid receptor-interacting protein 1) cDNA was cloned into the Xho I and BamH I sites of pET-15b.
3 shows that the GK2 sequence is inserted into the Nde I site in the pET-15b vector.
4A is a result of Western blot analysis of the level of the introduced protein after the cells were treated with Pep-1-PGAM1 fusion protein, GK2-PGAM1 fusion protein, or TAT-PGAM1 fusion protein for 20 minutes. Figure 4B shows the results of Western blot analysis of control PGAM1 protein, GK2-PGAM1 fusion protein, control CNRIP1 protein, and GK2-CNRIP1 fusion protein after cell introduction test.
Figure 5 is a photograph of the intracellular location and level of the cell-introduced Pep-1-PGAM1 fusion protein, GK2-PGAM1 fusion protein, and TAT-PGAM1 fusion protein with a confocal fluorescence microscope.
6 is a photograph of the intracellular location and level of the cell-introduced GK2-PGAM1 fusion protein or GK2-CNRIP1 fusion protein with a confocal fluorescence microscope.
7 is a result of testing the protective effect of the cell-introduced GK2-PGAM1 fusion protein or GK2-CNRIP1 fusion protein against oxidative stress. Cells were treated with 5 μM of GK2-PGAM1 fusion protein, GK2-CNRIP1 fusion protein, control PGAM1 protein, or control CNRIP1 protein for 1 hour and then treated with 100 μM H ₂ O ₂ for 1 hour. Thereafter, cell viability was evaluated by MTT assay.
8A, 8B, and 8C are results of testing the intracellular location and level of the cell-permeated GK2-PGAM1 fusion protein with a confocal fluorescence microscope. The median number of the fusion protein is the protein transport domain number shown in Table 1.
9 shows the protective effect of the cell-introduced GK2-PGAM1 fusion protein against oxidative stress. Cell viability was evaluated by MTT assay.

Hereinafter, the configuration of the present invention will be described in more detail with reference to specific embodiments. However, it is apparent to those of ordinary skill in the art that the scope of the present invention is not limited only to the description of the examples.

<Method>

1. Cell culture

NSC34 cells, a mouse motor neuron-like hybrid cell line, were distributed and cultured from Hallym University's College of Natural Sciences. The cell culture was prepared by adding 10% fetal bovine serum (FBS) and an antibiotic solution (100 units/ml penicillin, 100 μg/ml streptomycin) to DMEM medium. NSC34 cells were cultured using the above culture solution under conditions of maintaining 37°C, 95% humidity, and 5% CO ₂ , and when the cells adhered 70-80% to the culture dish, they were subcultured with trypsin-EDTA.

2. Construction of recombinant GK2

Annealing was performed at 37° C. for 2 hours using the following oligonucleotide.

Sense oligonucleotide (SEQ ID NO: 47):

tatgtacgcctaccgtatccgcgagttccgccagaccttccgcaagatcattcgcagcca,

Antisense oligonucleotide (SEQ ID NO: 48):

tatggctgcgaatgatcttgcggaaggtctggcggaactcgcggatacggtaggcgtaca.

Thereafter, the NdeI site of the pET-15b vector was cut, and the GK2 coding oligonucleotide pair prepared above was ligated. Recombined GK2 in Escherichia coli strain Top10 was transformed. DNA isolated from the transformed cells was amplified by PCR using the following primers.

Forward primer: T7, reverse primer: T7 terminator.

PGAM1 (Phosphoglycerate mutase 1) or CNRIP1 (CB1 cannabinoid receptor-interacting protein 1) gene was ligated by cutting the BamH Ⅰ and Xho Ⅰ sites of the GK2 vector.

3. Expression and purification of recombinant GK2

Recombinant GK2 plasmid was transferred to Escherichia coli It was transformed into strain BL21. Transformed cells were cultured in 100 ml LB medium containing 100 μg/ml ampicillin for 6 hours at 37° C. and 180 rpm for 6 hours, and then 0.5 mM IPTG (isopropyl-β-D-thiogalactoside) was added to 30° C., 120 rpm. Incubated for 16 hours. The recovered cells were sonicated and centrifuged to separate only the supernatant, and purified using a Ni ^{2 +} -nitrilotriacetic acid Sepharose affinity chromatography column. Protein concentration was normalized with bovine serum albumin (BSA) and quantified using a protein quantification kit.

4. NSC34 GK2 into cells Fusion protein Transduction

NSC34 cells were dispensed on a 60 mm plate and cultured at 37° C., 95% humidity, and 5% CO ₂ . In order to investigate the cellular permeability of GK2, the same concentration (5 μM) of PEP-1-PGAM1 fusion protein, TAT-PGAM1 fusion protein, and GK2-PGAM1 fusion protein were each treated for 20 minutes. In addition, in order to confirm that the cell permeability of the cell-penetrating peptide GK2 appears in any protein, another protein of the same concentration, GK2-CNRIP1 fusion protein, was treated for 20 minutes. The cultured cells were rinsed with PBS (phosphate buffer saline), and RIPA lysis buffer was added to extract proteins, followed by centrifugation (4° C., 12,000 rpm, 10 min) to quantify protein in the supernatant.

5. Western Blot analysis

50 μg of protein was mixed with 5X sample buffer and boiled for 2 minutes to prepare a protein sample, and then separated according to molecular weight using 15% mini gel SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis). After electrophoresis was completed, it was transferred to a nitrocellulose membrane, and blocking of the membrane was carried out at room temperature for 1 hour with TBS-T (pH 7.5, 25mM Tris-Cl, 150mM NaCl, 0.1% Tween 20) containing 5% skim milk. In order to measure the expression of the protein, the primary antibody (anti-histidine probe) was diluted 1:1,000 in TBS-T, reacted at room temperature for 1 hour, and washed with TBS-T. As a secondary antibody, HRP (horse radish peroxidase) conjugated anti-rabbit IgG was diluted 1:10,000 in TBS-T and reacted at room temperature for 1 hour. After washing with TBS-T, the expression level of each protein was confirmed using a detection reagent.

6. Fluorescence microscope analysis

Each glass cover slip was placed in a 24-well plate, and cells were dispensed thereon to incubate for 24 hours. 5 μM of Pep-1-PGAM1 fusion protein, TAT-PGAM1 fusion protein, GK2-PGAM1 fusion protein, or GK2-CNRIP1 fusion protein were transduced into NSC34 cells for 2 hours, and then the culture medium was removed and rinsed 3 times with PBS. . After 4% paraformaldehyde was added to each well and the cells were fixed for 5 minutes, PBS was added to wash the cells three times. After blocking and permeation in PBS (PBS-BT) containing 3% BSA and 0.1% Triton X-100 for 40 minutes at room temperature, it was washed 3 times with PBS-BT. His-probe primary antibody was diluted 1:2,000 and incubated at room temperature for 1 hour. The secondary antibody (Alexa Fluor 488) was diluted to 1:10,000, darkened, reacted for 1 hour, and washed 3 times for 5 minutes with PBS-BT. The glass cover slip was removed from the well to remove moisture from the edge and lifted. It was observed with a confocal fluorescence microscope and fluorescence expression was confirmed using an image analyzer.

<Result>

Outcome 1: GK2 Fusion protein Schematic diagram and refinement

In order to express the cell-penetrating peptide, the GK2 oligonucleotide (tacgcctac cgtatccgcg agttccgcca gaccttccgc aagatcattc gcagc) was cloned into the pET-15b plasmid Nde I site [Fig. 3]. To determine the cell penetration efficiency of a cell-permeable peptide GK2 the Xho I and BamH I sites of the pET-15b plasmid PGAM1 (Phosphoglycerate mutase 1) cDNA [1] or CNRIP1 (CB1 cannabinoid receptor-interacting protein 1) cDNA [ 2 ] Was cloned. As a control vector, only PGAM1 or CNRIP1 was cloned into the pET-15b plasmid. After expression of the fusion protein, it was purified using a Ni ²⁺ -nitrilotriacetic acid Sepharose affinity chromatography column, which was confirmed by SDS-PAGE [Fig. 1, Fig. 2].

Result 2: GK2 Fusion protein Cell introduction

In order to examine the cell permeation efficiency of GK2, the same concentration (5 μM) of PEP-1-PGAM1 fusion protein, GK2-PGAM1 fusion protein, or Tat-PGAM1 fusion protein was treated on NSC34 cells for 20 minutes. The cell permeation level was then analyzed by Western blotting. The level of intracellular fusion protein was highest in GK2-PGAM1 fusion protein, compared to that of TAT-PGAM1 fusion protein, about 90%, and PEP-1-PGAM1 fusion protein, which showed about 38% cell penetration level [Fig. 4A] . The control PGAM1 protein was not penetrated into the cells.

In order to prove that the cell-penetrating peptide GK2 fuses with an arbitrary protein and penetrates into the cell, the GK2-PGAM1 fusion protein and GK2-CNRIP1 fusion protein prepared by fusion of the cell-penetrating peptide GK2 to two types of proteins PGAM1 and CNRIP1 are identical. The cells were treated for 20 minutes at a concentration (5 μM). Although there is a difference in cell permeation efficiency according to the protein type, the two types of proteins were smoothly permeated into the NSC34 cells [Fig. 4B].

To determine the location of the Pep-1-PGAM1 fusion protein, GK2-PGAM1 fusion protein, or TAT-PGAM1 fusion protein introduced into the cell, each fusion protein-permeated cell was immunostained with DAPI and Alexa Fluor 488-conjugated secondary antibody. I did. Control PGAM1 protein was not observed in NSC34 cells. Unlike this, PEP-1-PGAM1 fusion protein, GK2-PGAM1 fusion protein, or TAT-PGAM1 fusion protein was detected mainly in the cytoplasm by fluorescence microscopy, and also in the nucleus [Fig. 5]. In addition, the GK2-PGAM1 fusion protein exhibited a higher intensity fluorescence signal than the PEP-1-PGAM1 fusion protein and the TAT-PGAM1 fusion protein. This suggests that the cell penetration efficiency of the GK2-PGAM1 fusion protein is the highest.

In addition, the intracellular location of GK2 transmitted through fusion with an arbitrary protein was investigated in the same manner as above. The GK2-PGAM1 fusion protein and the GK2-CNRIP1 fusion protein were mainly detected in the cytoplasm and also in the nucleus [Fig. 6].

Outcome 3: GK2 for oxidative stress Fusion protein effect

It was investigated whether the GK2 fusion protein penetrated into cells has a protective effect against oxidative stress. A constant concentration (5 μM) of GK2-PGAM1 fusion protein, GK2-CNRIP1 fusion protein, control PGAM1 protein, and control CNRIP1 protein were treated for 1 hour and then treated with 100 μM H ₂ O ₂ for 1 hour. Then, the cell viability was evaluated by MTT assay [FIG. 7]. The cell viability of the cells treated with H ₂ O ₂ decreased by about 60% compared to the control cells. The survival rates of cells pretreated with GK2-PGAM1 and GK2-CNRIP1 increased to about 80% and 88%. In comparison, the control PGAM1 protein and the control CNRIP1 protein had no protective effect on cell death by H ₂ O ₂ .

Outcome 4: GK2 cells permeability Peptide Follow Fusion protein Effects on cell introduction and oxidative stress

Table 1 shows GK2 derivative peptides (GK2-1 to GK2-21) in which one or more peptides are substituted or added to the GK2 peptide sequence (RAYRIREFRQTFRKIIRS). Immunostaining was performed in the same manner as above to confirm the intracellular location of the GK2 derivative peptide and the PGAM1 fusion protein that penetrated the cell. GK2 candidate proteins were also detected in the cytoplasm and nucleus. Among them, 8 cells having good cell permeation efficiency were selected and shown in FIG. 8.

MTT analysis was performed in the same manner as above to determine the cytoprotective effect of the fusion protein fused with the GK2 derivative peptide against oxidative stress. The cell viability, which was reduced by 60% by H ₂ O ₂ , increased from 72% to a maximum of 87% when the fusion protein fused with the GK2 candidate protein and the PGAM1 protein was pretreated [Fig. 9]. In the case of the GK2-PGAM1 fusion protein, the cell viability increased to 88%.

Table 1 below shows the amino acid sequences of 23 specific examples among the cell-penetrating peptides of the present invention, and are respectively SEQ ID NOs: 1 to 23 in the order of the table. In addition, Table 2 below is a nucleotide sequence encoding 23 specific examples among the cell-penetrating peptides of the present invention, and are respectively SEQ ID NOs: 24 to 46 in the order of the table. The amino acid sequence of these cell-penetrating peptides (SEQ ID NO: 1-23) and the oligonucleotide sequence (SEQ ID NO: 24-46) encoding these cell-penetrating peptides are only representative examples, and the amino acid sequence of the cell-penetrating peptide and the oligonucleotide sequence encoding the same This is not limited thereto is apparent to those of ordinary skill in the art to which the present invention belongs.

In addition, the amino acid sequence of the cell-penetrating phosphoglycerate mutase 1 fusion protein bound to the GK2 to GK2-21 cell-penetrating peptide of Table 1 below is in sequence SEQ ID NO: 49 to 71, respectively, and these cell-penetrating phosphoglycerate mutases 1 The recombinant oligonucleotide sequence encoding the fusion protein is shown in the sequence listing as SEQ ID NOs: 72 to 94, respectively.

These fusion protein amino acid sequences and recombinant oligonucleotide sequences are representative examples, but the amino acid sequence of the cell-penetrating phosphoglycerate mutase 1 fusion protein and the recombinant oligonucleotide sequence encoding the fusion protein are not limited thereto. It is obvious to those of ordinary skill in the technical field.

Name Peptide sequence GK2 YAYRIREFRQTFRKIIRS GK2-0 RAYRIREFRQTFRKIIRS GK2-1 RRYRIREFRQTFRKIIRS GK2-2 RRRRIREFRQTFRKIIRS GK2-3 YAYRIREFRQTFRKIIRR GK2-4 YAYRIREFRQTFRKIRRR GK2-5 YAYRIREFRQTFRKRRRR GK2-6 YARRIREFRQTFRKIIRS GK2-7 YAYRIREFRQTFRKIRRS GK2-8 YAYRIREFRQTFRKRIRS GK2-9 RYAYRIREFRQTFRKIIRS GK2-10 RRYAYRIREFRQTFRKIIRS GK2-11 RRRYAYRIREFRQTFRKIIRS GK2-12 YAYRIREFRQTFRKIIRSR GK2-13 YAYRIREFRQTFRKIIRSRR GK2-14 YAYRIREFRQTFRKIIRSRRR GK2-15 RKKRRQRRRYAYRIREFRQTFRKIIRS GK2-16 RKKRRQRRRIREFRQTFRKIIRS GK2-17 RKKRRQRRREFRQTFRKIIRS GK2-18 YAYRIREFRQTFRKIIRKKRRQRRR GK2-19 YAYRIREFRQTFRKKRRQRRR GK2-20 KETWWETWWTEWSQPKKKRKVRQTFRKIIRS GK2-21 YAYRIREFKETWWETWWTEWSQPKKKRKV

Name Nucleotide sequence GK2 tac gcc tac cgt atc cgc gag ttc cgc cag acc ttc cgc aag atc att cgc agc GK2-0 cgc gcc tac cgt atc cgc gag ttc cgc cag acc ttc cgc aag atc att cgc agc GK2-1 cgc cgt tac cgt atc cgc gag ttc cgc cag acc ttc cgc aag atc att cgc agc GK2-2 cgc cgt cgc cgt atc cgc gag ttc cgc cag acc ttc cgc aag atc att cgc agc GK2-3 tac gcc tac cgt atc cgc gag ttc cgc cag acc ttc cgc aag atc att cgc cgc GK2-4 tac gcc tac cgt atc cgc gag ttc cgc cag acc ttc cgc aag atc cgt cgc cgc GK2-5 tac gcc tac cgt atc cgc gag ttc cgc cag acc ttc cgc aag cgc cgt cgc cgc GK2-6 tac gcc cgc cgt atc cgc gag ttc cgc cag acc ttc cgc aag atc att cgc agc GK2-7 tac gcc tac cgt atc cgc gag ttc cgc cag acc ttc cgc aag atc cgc cgc agc GK2-8 tac gcc tac cgt atc cgc gag ttc cgc cag acc ttc cgc aag cgc att cgc agc GK2-9 cgc tac gcc tac cgt atc cgc gag ttc cgc cag acc ttc cgc aag atc att cgc agc GK2-10 cgc cgt tac gcc tac cgt atc cgc gag ttc cgc cag acc ttc cgc aag atc att cgc agc GK2-11 cgc cgt cgc tac gcc tac cgt atc cgc gag ttc cgc cag acc ttc cgc aag atc att cgc agc GK2-12 tac gcc tac cgt atc cgc gag ttc cgc cag acc ttc cgc aag atc att cgc agc cgc GK2-13 tac gcc tac cgt atc cgc gag ttc cgc cag acc ttc cgc aag atc att cgc agc cgc cgt GK2-14 tac gcc tac cgt atc cgc gag ttc cgc cag acc ttc cgc aag atc att cgc agc cgc cgt cgc GK2-15 agg aag aag cgg aga cag cga cga aga tac gcc tac cgt atc cgc gag ttc cgc cag acc ttc cgc aag atc att cgc agc GK2-16 agg aag aag cgg aga cag cga cga aga atc cgc gag ttc cgc cag acc ttc cgc aag atc att cgc agc GK2-17 agg aag aag cgg aga cag cga cga aga gag ttc cgc cag acc ttc cgc aag atc att cgc agc GK2-18 tac gcc tac cgt atc cgc gag ttc cgc cag acc ttc cgc aag atc att cgc aag aag cgg aga cag cga cga aga GK2-19 tac gcc tac cgt atc cgc gag ttc cgc cag acc ttc cgc aag aag cgg aga cag cga cga aga GK2-20 aaa gaa acc tgg tgg gaa acc tgg tgg acc gaa tgg tct cag ccg aaa aaa aaa cgt aaa gtg cgc cag acc ttc cgc aag atc att cgc agc GK2-21 tac gcc tac cgt atc cgc gag ttc aaa gaa acc tgg tgg gaa acc tgg tgg acc gaa tgg tct cag ccg aaa aaa aaa cgt aaa gtg

<110> Industry Academic Cooperation Foundation, Hallym University <120> Cell-penetrating phosphoglycerate mutase 1 fusion protein and pharmaceutical Composition containing thereof <130> HallymU-SYCHOI-GK2-PGAM1-190509 <160> 94 <170> KoPatentIn 3.0 <210> 1 <211> 18 <212> PRT <213> Homo Sapiens <400> 1 Tyr Ala Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Ile Ile 1 5 10 15 Arg Ser <210> 2 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> novel cell-penetrating peptide <400> 2 Arg Ala Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Ile Ile 1 5 10 15 Arg Ser <210> 3 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> novel cell-penetrating peptide <400> 3 Arg Arg Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Ile Ile 1 5 10 15 Arg Ser <210> 4 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> novel cell-penetrating peptide <400> 4 Arg Arg Arg Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Ile Ile 1 5 10 15 Arg Ser <210> 5 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> novel cell-penetrating peptide <400> 5 Tyr Ala Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Ile Ile 1 5 10 15 Arg Arg <210> 6 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> novel cell-penetrating peptide <400> 6 Tyr Ala Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Ile Arg 1 5 10 15 Arg Arg <210> 7 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> novel cell-penetrating peptide <400> 7 Tyr Ala Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Arg Arg 1 5 10 15 Arg Arg <210> 8 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> novel cell-penetrating peptide <400> 8 Tyr Ala Arg Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Ile Ile 1 5 10 15 Arg Ser <210> 9 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> novel cell-penetrating peptide <400> 9 Tyr Ala Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Ile Arg 1 5 10 15 Arg Ser <210> 10 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> novel cell-penetrating peptide <400> 10 Tyr Ala Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Arg Ile 1 5 10 15 Arg Ser <210> 11 <211> 19 <212> PRT <213> Artificial Sequence <220> <223> novel cell-penetrating peptide <400> 11 Arg Tyr Ala Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Ile 1 5 10 15 Ile Arg Ser <210> 12 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> novel cell-penetrating peptide <400> 12 Arg Arg Tyr Ala Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys 1 5 10 15 Ile Ile Arg Ser 20 <210> 13 <211> 21 <212> PRT <213> Artificial Sequence <220> <223> novel cell-penetrating peptide <400> 13 Arg Arg Arg Tyr Ala Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg 1 5 10 15 Lys Ile Ile Arg Ser 20 <210> 14 <211> 19 <212> PRT <213> Artificial Sequence <220> <223> novel cell-penetrating peptide <400> 14 Tyr Ala Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Ile Ile 1 5 10 15 Arg Ser Arg <210> 15 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> novel cell-penetrating peptide <400> 15 Tyr Ala Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Ile Ile 1 5 10 15 Arg Ser Arg Arg 20 <210> 16 <211> 21 <212> PRT <213> Artificial Sequence <220> <223> novel cell-penetrating peptide <400> 16 Tyr Ala Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Ile Ile 1 5 10 15 Arg Ser Arg Arg Arg 20 <210> 17 <211> 27 <212> PRT <213> Artificial Sequence <220> <223> novel cell-penetrating peptide <400> 17 Arg Lys Lys Arg Arg Gln Arg Arg Arg Tyr Ala Tyr Arg Ile Arg Glu 1 5 10 15 Phe Arg Gln Thr Phe Arg Lys Ile Ile Arg Ser 20 25 <210> 18 <211> 23 <212> PRT <213> Artificial Sequence <220> <223> novel cell-penetrating peptide <400> 18 Arg Lys Lys Arg Arg Gln Arg Arg Arg Ile Arg Glu Phe Arg Gln Thr 1 5 10 15 Phe Arg Lys Ile Ile Arg Ser 20 <210> 19 <211> 21 <212> PRT <213> Artificial Sequence <220> <223> novel cell-penetrating peptide <400> 19 Arg Lys Lys Arg Arg Gln Arg Arg Arg Glu Phe Arg Gln Thr Phe Arg 1 5 10 15 Lys Ile Ile Arg Ser 20 <210> 20 <211> 25 <212> PRT <213> Artificial Sequence <220> <223> novel cell-penetrating peptide <400> 20 Tyr Ala Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Ile Ile 1 5 10 15 Arg Lys Lys Arg Arg Gln Arg Arg Arg 20 25 <210> 21 <211> 21 <212> PRT <213> Artificial Sequence <220> <223> novel cell-penetrating peptide <400> 21 Tyr Ala Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Lys Arg 1 5 10 15 Arg Gln Arg Arg Arg 20 <210> 22 <211> 31 <212> PRT <213> Artificial Sequence <220> <223> novel cell-penetrating peptide <400> 22 Lys Glu Thr Trp Trp Glu Thr Trp Trp Thr Glu Trp Ser Gln Pro Lys 1 5 10 15 Lys Lys Arg Lys Val Arg Gln Thr Phe Arg Lys Ile Ile Arg Ser 20 25 30 <210> 23 <211> 29 <212> PRT <213> Artificial Sequence <220> <223> novel cell-penetrating peptide <400> 23 Tyr Ala Tyr Arg Ile Arg Glu Phe Lys Glu Thr Trp Trp Glu Thr Trp 1 5 10 15 Trp Thr Glu Trp Ser Gln Pro Lys Lys Lys Arg Lys Val 20 25 <210> 24 <211> 54 <212> DNA <213> Homo Sapiens <400> 24 tacgcctacc gtatccgcga gttccgccag accttccgca agatcattcg cagc 54 <210> 25 <211> 54 <212> DNA <213> Artificial Sequence <220> <223> cell-penetrating peptide coding sequence <400> 25 cgcgcctacc gtatccgcga gttccgccag accttccgca agatcattcg cagc 54 <210> 26 <211> 54 <212> DNA <213> Artificial Sequence <220> <223> cell-penetrating peptide coding sequence <400> 26 cgccgttacc gtatccgcga gttccgccag accttccgca agatcattcg cagc 54 <210> 27 <211> 54 <212> DNA <213> Artificial Sequence <220> <223> cell-penetrating peptide coding sequence <400> 27 cgccgtcgcc gtatccgcga gttccgccag accttccgca agatcattcg cagc 54 <210> 28 <211> 54 <212> DNA <213> Artificial Sequence <220> <223> cell-penetrating peptide coding sequence <400> 28 tacgcctacc gtatccgcga gttccgccag accttccgca agatcattcg ccgc 54 <210> 29 <211> 54 <212> DNA <213> Artificial Sequence <220> <223> cell-penetrating peptide coding sequence <400> 29 tacgcctacc gtatccgcga gttccgccag accttccgca agatccgtcg ccgc 54 <210> 30 <211> 54 <212> DNA <213> Artificial Sequence <220> <223> cell-penetrating peptide coding sequence <400> 30 tacgcctacc gtatccgcga gttccgccag accttccgca agcgccgtcg ccgc 54 <210> 31 <211> 54 <212> DNA <213> Artificial Sequence <220> <223> cell-penetrating peptide coding sequence <400> 31 tacgcccgcc gtatccgcga gttccgccag accttccgca agatcattcg cagc 54 <210> 32 <211> 54 <212> DNA <213> Artificial Sequence <220> <223> cell-penetrating peptide coding sequence <400> 32 tacgcctacc gtatccgcga gttccgccag accttccgca agatccgccg cagc 54 <210> 33 <211> 54 <212> DNA <213> Artificial Sequence <220> <223> cell-penetrating peptide coding sequence <400> 33 tacgcctacc gtatccgcga gttccgccag accttccgca agcgcattcg cagc 54 <210> 34 <211> 57 <212> DNA <213> Artificial Sequence <220> <223> cell-penetrating peptide coding sequence <400> 34 cgctacgcct accgtatccg cgagttccgc cagaccttcc gcaagatcat tcgcagc 57 <210> 35 <211> 60 <212> DNA <213> Artificial Sequence <220> <223> cell-penetrating peptide coding sequence <400> 35 cgccgttacg cctaccgtat ccgcgagttc cgccagacct tccgcaagat cattcgcagc 60 60 <210> 36 <211> 63 <212> DNA <213> Artificial Sequence <220> <223> cell-penetrating peptide coding sequence <400> 36 cgccgtcgct acgcctaccg tatccgcgag ttccgccaga ccttccgcaa gatcattcgc 60 agc 63 <210> 37 <211> 57 <212> DNA <213> Artificial Sequence <220> <223> cell-penetrating peptide coding sequence <400> 37 tacgcctacc gtatccgcga gttccgccag accttccgca agatcattcg cagccgc 57 <210> 38 <211> 60 <212> DNA <213> Artificial Sequence <220> <223> cell-penetrating peptide coding sequence <400> 38 tacgcctacc gtatccgcga gttccgccag accttccgca agatcattcg cagccgccgt 60 60 <210> 39 <211> 63 <212> DNA <213> Artificial Sequence <220> <223> cell-penetrating peptide coding sequence <400> 39 tacgcctacc gtatccgcga gttccgccag accttccgca agatcattcg cagccgccgt 60 cgc 63 <210> 40 <211> 81 <212> DNA <213> Artificial Sequence <220> <223> cell-penetrating peptide coding sequence <400> 40 aggaagaagc ggagacagcg acgaagatac gcctaccgta tccgcgagtt ccgccagacc 60 ttccgcaaga tcattcgcag c 81 <210> 41 <211> 69 <212> DNA <213> Artificial Sequence <220> <223> cell-penetrating peptide coding sequence <400> 41 aggaagaagc ggagacagcg acgaagaatc cgcgagttcc gccagacctt ccgcaagatc 60 attcgcagc 69 <210> 42 <211> 63 <212> DNA <213> Artificial Sequence <220> <223> cell-penetrating peptide coding sequence <400> 42 aggaagaagc ggagacagcg acgaagagag ttccgccaga ccttccgcaa gatcattcgc 60 agc 63 <210> 43 <211> 75 <212> DNA <213> Artificial Sequence <220> <223> cell-penetrating peptide coding sequence <400> 43 tacgcctacc gtatccgcga gttccgccag accttccgca agatcattcg caagaagcgg 60 agacagcgac gaaga 75 <210> 44 <211> 63 <212> DNA <213> Artificial Sequence <220> <223> cell-penetrating peptide coding sequence <400> 44 tacgcctacc gtatccgcga gttccgccag accttccgca agaagcggag acagcgacga 60 aga 63 <210> 45 <211> 93 <212> DNA <213> Artificial Sequence <220> <223> cell-penetrating peptide coding sequence <400> 45 aaagaaacct ggtgggaaac ctggtggacc gaatggtctc agccgaaaaa aaaacgtaaa 60 gtgcgccaga ccttccgcaa gatcattcgc agc 93 <210> 46 <211> 87 <212> DNA <213> Artificial Sequence <220> <223> cell-penetrating peptide coding sequence <400> 46 tacgcctacc gtatccgcga gttcaaagaa acctggtggg aaacctggtg gaccgaatgg 60 tctcagccga aaaaaaaacg taaagtg 87 <210> 47 <211> 60 <212> DNA <213> Artificial Sequence <220> <223> sense oligonucleotide for cell-penetrating peptide <400> 47 tatgtacgcc taccgtatcc gcgagttccg ccagaccttc cgcaagatca ttcgcagcca 60 60 <210> 48 <211> 60 <212> DNA <213> Artificial Sequence <220> <223> antisense oligonucleotide for cell-penetrating peptide <400> 48 tatggctgcg aatgatcttg cggaaggtct ggcggaactc gcggatacgg taggcgtaca 60 60 <210> 49 <211> 276 <212> PRT <213> Artificial Sequence <220> <223> GK2-PGAM1 fusion protein <400> 49 Tyr Ala Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Ile Ile 1 5 10 15 Arg Ser His Met Leu Glu Met Ala Ala Tyr Lys Leu Val Leu Ile Arg 20 25 30 His Gly Glu Ser Ala Trp Asn Leu Glu Asn Arg Phe Ser Gly Trp Tyr 35 40 45 Asp Ala Asp Leu Ser Pro Ala Gly His Glu Glu Ala Lys Arg Gly Gly 50 55 60 Gln Ala Leu Arg Asp Ala Gly Tyr Glu Phe Asp Ile Cys Phe Thr Ser 65 70 75 80 Val Gln Lys Arg Ala Ile Arg Thr Leu Trp Thr Val Leu Asp Ala Ile 85 90 95 Asp Gln Met Trp Leu Pro Val Val Arg Thr Trp Arg Leu Asn Glu Arg 100 105 110 His Tyr Gly Gly Leu Thr Gly Leu Asn Lys Ala Glu Thr Ala Ala Lys 115 120 125 His Gly Glu Ala Gln Val Lys Ile Trp Arg Arg Ser Tyr Asp Val Pro 130 135 140 Pro Pro Pro Met Glu Pro Asp His Pro Phe Tyr Ser Asn Ile Ser Lys 145 150 155 160 Asp Arg Arg Tyr Ala Asp Leu Thr Glu Asp Gln Leu Pro Ser Cys Glu 165 170 175 Ser Leu Lys Asp Thr Ile Ala Arg Ala Leu Pro Phe Trp Asn Glu Glu 180 185 190 Ile Val Pro Gln Ile Lys Glu Gly Lys Arg Val Leu Ile Ala Ala His 195 200 205 Gly Asn Ser Leu Arg Gly Ile Val Lys His Leu Glu Gly Leu Ser Glu 210 215 220 Glu Ala Ile Met Glu Leu Asn Leu Pro Thr Gly Ile Pro Ile Val Tyr 225 230 235 240 Glu Leu Asp Lys Asn Leu Lys Pro Ile Lys Pro Met Gln Phe Leu Gly 245 250 255 Met Lys Arg Arg Cys Ala Lys Pro Trp Lys Ala Val Ala Ala Gln Gly 260 265 270 Lys Ala Lys Lys 275 <210> 50 <211> 276 <212> PRT <213> Artificial Sequence <220> <223> GK2-0-PGAM1 fusion protein <400> 50 Arg Ala Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Ile Ile 1 5 10 15 Arg Ser His Met Leu Glu Met Ala Ala Tyr Lys Leu Val Leu Ile Arg 20 25 30 His Gly Glu Ser Ala Trp Asn Leu Glu Asn Arg Phe Ser Gly Trp Tyr 35 40 45 Asp Ala Asp Leu Ser Pro Ala Gly His Glu Glu Ala Lys Arg Gly Gly 50 55 60 Gln Ala Leu Arg Asp Ala Gly Tyr Glu Phe Asp Ile Cys Phe Thr Ser 65 70 75 80 Val Gln Lys Arg Ala Ile Arg Thr Leu Trp Thr Val Leu Asp Ala Ile 85 90 95 Asp Gln Met Trp Leu Pro Val Val Arg Thr Trp Arg Leu Asn Glu Arg 100 105 110 His Tyr Gly Gly Leu Thr Gly Leu Asn Lys Ala Glu Thr Ala Ala Lys 115 120 125 His Gly Glu Ala Gln Val Lys Ile Trp Arg Arg Ser Tyr Asp Val Pro 130 135 140 Pro Pro Pro Met Glu Pro Asp His Pro Phe Tyr Ser Asn Ile Ser Lys 145 150 155 160 Asp Arg Arg Tyr Ala Asp Leu Thr Glu Asp Gln Leu Pro Ser Cys Glu 165 170 175 Ser Leu Lys Asp Thr Ile Ala Arg Ala Leu Pro Phe Trp Asn Glu Glu 180 185 190 Ile Val Pro Gln Ile Lys Glu Gly Lys Arg Val Leu Ile Ala Ala His 195 200 205 Gly Asn Ser Leu Arg Gly Ile Val Lys His Leu Glu Gly Leu Ser Glu 210 215 220 Glu Ala Ile Met Glu Leu Asn Leu Pro Thr Gly Ile Pro Ile Val Tyr 225 230 235 240 Glu Leu Asp Lys Asn Leu Lys Pro Ile Lys Pro Met Gln Phe Leu Gly 245 250 255 Met Lys Arg Arg Cys Ala Lys Pro Trp Lys Ala Val Ala Ala Gln Gly 260 265 270 Lys Ala Lys Lys 275 <210> 51 <211> 276 <212> PRT <213> Artificial Sequence <220> <223> GK2-1-PGAM1 fusion protein <400> 51 Arg Arg Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Ile Ile 1 5 10 15 Arg Ser His Met Leu Glu Met Ala Ala Tyr Lys Leu Val Leu Ile Arg 20 25 30 His Gly Glu Ser Ala Trp Asn Leu Glu Asn Arg Phe Ser Gly Trp Tyr 35 40 45 Asp Ala Asp Leu Ser Pro Ala Gly His Glu Glu Ala Lys Arg Gly Gly 50 55 60 Gln Ala Leu Arg Asp Ala Gly Tyr Glu Phe Asp Ile Cys Phe Thr Ser 65 70 75 80 Val Gln Lys Arg Ala Ile Arg Thr Leu Trp Thr Val Leu Asp Ala Ile 85 90 95 Asp Gln Met Trp Leu Pro Val Val Arg Thr Trp Arg Leu Asn Glu Arg 100 105 110 His Tyr Gly Gly Leu Thr Gly Leu Asn Lys Ala Glu Thr Ala Ala Lys 115 120 125 His Gly Glu Ala Gln Val Lys Ile Trp Arg Arg Ser Tyr Asp Val Pro 130 135 140 Pro Pro Pro Met Glu Pro Asp His Pro Phe Tyr Ser Asn Ile Ser Lys 145 150 155 160 Asp Arg Arg Tyr Ala Asp Leu Thr Glu Asp Gln Leu Pro Ser Cys Glu 165 170 175 Ser Leu Lys Asp Thr Ile Ala Arg Ala Leu Pro Phe Trp Asn Glu Glu 180 185 190 Ile Val Pro Gln Ile Lys Glu Gly Lys Arg Val Leu Ile Ala Ala His 195 200 205 Gly Asn Ser Leu Arg Gly Ile Val Lys His Leu Glu Gly Leu Ser Glu 210 215 220 Glu Ala Ile Met Glu Leu Asn Leu Pro Thr Gly Ile Pro Ile Val Tyr 225 230 235 240 Glu Leu Asp Lys Asn Leu Lys Pro Ile Lys Pro Met Gln Phe Leu Gly 245 250 255 Met Lys Arg Arg Cys Ala Lys Pro Trp Lys Ala Val Ala Ala Gln Gly 260 265 270 Lys Ala Lys Lys 275 <210> 52 <211> 276 <212> PRT <213> Artificial Sequence <220> <223> GK2-2-PGAM1 fusion protein <400> 52 Arg Arg Arg Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Ile Ile 1 5 10 15 Arg Ser His Met Leu Glu Met Ala Ala Tyr Lys Leu Val Leu Ile Arg 20 25 30 His Gly Glu Ser Ala Trp Asn Leu Glu Asn Arg Phe Ser Gly Trp Tyr 35 40 45 Asp Ala Asp Leu Ser Pro Ala Gly His Glu Glu Ala Lys Arg Gly Gly 50 55 60 Gln Ala Leu Arg Asp Ala Gly Tyr Glu Phe Asp Ile Cys Phe Thr Ser 65 70 75 80 Val Gln Lys Arg Ala Ile Arg Thr Leu Trp Thr Val Leu Asp Ala Ile 85 90 95 Asp Gln Met Trp Leu Pro Val Val Arg Thr Trp Arg Leu Asn Glu Arg 100 105 110 His Tyr Gly Gly Leu Thr Gly Leu Asn Lys Ala Glu Thr Ala Ala Lys 115 120 125 His Gly Glu Ala Gln Val Lys Ile Trp Arg Arg Ser Tyr Asp Val Pro 130 135 140 Pro Pro Pro Met Glu Pro Asp His Pro Phe Tyr Ser Asn Ile Ser Lys 145 150 155 160 Asp Arg Arg Tyr Ala Asp Leu Thr Glu Asp Gln Leu Pro Ser Cys Glu 165 170 175 Ser Leu Lys Asp Thr Ile Ala Arg Ala Leu Pro Phe Trp Asn Glu Glu 180 185 190 Ile Val Pro Gln Ile Lys Glu Gly Lys Arg Val Leu Ile Ala Ala His 195 200 205 Gly Asn Ser Leu Arg Gly Ile Val Lys His Leu Glu Gly Leu Ser Glu 210 215 220 Glu Ala Ile Met Glu Leu Asn Leu Pro Thr Gly Ile Pro Ile Val Tyr 225 230 235 240 Glu Leu Asp Lys Asn Leu Lys Pro Ile Lys Pro Met Gln Phe Leu Gly 245 250 255 Met Lys Arg Arg Cys Ala Lys Pro Trp Lys Ala Val Ala Ala Gln Gly 260 265 270 Lys Ala Lys Lys 275 <210> 53 <211> 276 <212> PRT <213> Artificial Sequence <220> <223> GK2-3-PGAM1 fusion protein <400> 53 Tyr Ala Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Ile Ile 1 5 10 15 Arg Arg His Met Leu Glu Met Ala Ala Tyr Lys Leu Val Leu Ile Arg 20 25 30 His Gly Glu Ser Ala Trp Asn Leu Glu Asn Arg Phe Ser Gly Trp Tyr 35 40 45 Asp Ala Asp Leu Ser Pro Ala Gly His Glu Glu Ala Lys Arg Gly Gly 50 55 60 Gln Ala Leu Arg Asp Ala Gly Tyr Glu Phe Asp Ile Cys Phe Thr Ser 65 70 75 80 Val Gln Lys Arg Ala Ile Arg Thr Leu Trp Thr Val Leu Asp Ala Ile 85 90 95 Asp Gln Met Trp Leu Pro Val Val Arg Thr Trp Arg Leu Asn Glu Arg 100 105 110 His Tyr Gly Gly Leu Thr Gly Leu Asn Lys Ala Glu Thr Ala Ala Lys 115 120 125 His Gly Glu Ala Gln Val Lys Ile Trp Arg Arg Ser Tyr Asp Val Pro 130 135 140 Pro Pro Pro Met Glu Pro Asp His Pro Phe Tyr Ser Asn Ile Ser Lys 145 150 155 160 Asp Arg Arg Tyr Ala Asp Leu Thr Glu Asp Gln Leu Pro Ser Cys Glu 165 170 175 Ser Leu Lys Asp Thr Ile Ala Arg Ala Leu Pro Phe Trp Asn Glu Glu 180 185 190 Ile Val Pro Gln Ile Lys Glu Gly Lys Arg Val Leu Ile Ala Ala His 195 200 205 Gly Asn Ser Leu Arg Gly Ile Val Lys His Leu Glu Gly Leu Ser Glu 210 215 220 Glu Ala Ile Met Glu Leu Asn Leu Pro Thr Gly Ile Pro Ile Val Tyr 225 230 235 240 Glu Leu Asp Lys Asn Leu Lys Pro Ile Lys Pro Met Gln Phe Leu Gly 245 250 255 Met Lys Arg Arg Cys Ala Lys Pro Trp Lys Ala Val Ala Ala Gln Gly 260 265 270 Lys Ala Lys Lys 275 <210> 54 <211> 276 <212> PRT <213> Artificial Sequence <220> <223> GK2-4-PGAM1 fusion protein <400> 54 Tyr Ala Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Ile Arg 1 5 10 15 Arg Arg His Met Leu Glu Met Ala Ala Tyr Lys Leu Val Leu Ile Arg 20 25 30 His Gly Glu Ser Ala Trp Asn Leu Glu Asn Arg Phe Ser Gly Trp Tyr 35 40 45 Asp Ala Asp Leu Ser Pro Ala Gly His Glu Glu Ala Lys Arg Gly Gly 50 55 60 Gln Ala Leu Arg Asp Ala Gly Tyr Glu Phe Asp Ile Cys Phe Thr Ser 65 70 75 80 Val Gln Lys Arg Ala Ile Arg Thr Leu Trp Thr Val Leu Asp Ala Ile 85 90 95 Asp Gln Met Trp Leu Pro Val Val Arg Thr Trp Arg Leu Asn Glu Arg 100 105 110 His Tyr Gly Gly Leu Thr Gly Leu Asn Lys Ala Glu Thr Ala Ala Lys 115 120 125 His Gly Glu Ala Gln Val Lys Ile Trp Arg Arg Ser Tyr Asp Val Pro 130 135 140 Pro Pro Pro Met Glu Pro Asp His Pro Phe Tyr Ser Asn Ile Ser Lys 145 150 155 160 Asp Arg Arg Tyr Ala Asp Leu Thr Glu Asp Gln Leu Pro Ser Cys Glu 165 170 175 Ser Leu Lys Asp Thr Ile Ala Arg Ala Leu Pro Phe Trp Asn Glu Glu 180 185 190 Ile Val Pro Gln Ile Lys Glu Gly Lys Arg Val Leu Ile Ala Ala His 195 200 205 Gly Asn Ser Leu Arg Gly Ile Val Lys His Leu Glu Gly Leu Ser Glu 210 215 220 Glu Ala Ile Met Glu Leu Asn Leu Pro Thr Gly Ile Pro Ile Val Tyr 225 230 235 240 Glu Leu Asp Lys Asn Leu Lys Pro Ile Lys Pro Met Gln Phe Leu Gly 245 250 255 Met Lys Arg Arg Cys Ala Lys Pro Trp Lys Ala Val Ala Ala Gln Gly 260 265 270 Lys Ala Lys Lys 275 <210> 55 <211> 276 <212> PRT <213> Artificial Sequence <220> <223> GK2-5-PGAM1 fusion protein <400> 55 Tyr Ala Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Arg Arg 1 5 10 15 Arg Arg His Met Leu Glu Met Ala Ala Tyr Lys Leu Val Leu Ile Arg 20 25 30 His Gly Glu Ser Ala Trp Asn Leu Glu Asn Arg Phe Ser Gly Trp Tyr 35 40 45 Asp Ala Asp Leu Ser Pro Ala Gly His Glu Glu Ala Lys Arg Gly Gly 50 55 60 Gln Ala Leu Arg Asp Ala Gly Tyr Glu Phe Asp Ile Cys Phe Thr Ser 65 70 75 80 Val Gln Lys Arg Ala Ile Arg Thr Leu Trp Thr Val Leu Asp Ala Ile 85 90 95 Asp Gln Met Trp Leu Pro Val Val Arg Thr Trp Arg Leu Asn Glu Arg 100 105 110 His Tyr Gly Gly Leu Thr Gly Leu Asn Lys Ala Glu Thr Ala Ala Lys 115 120 125 His Gly Glu Ala Gln Val Lys Ile Trp Arg Arg Ser Tyr Asp Val Pro 130 135 140 Pro Pro Pro Met Glu Pro Asp His Pro Phe Tyr Ser Asn Ile Ser Lys 145 150 155 160 Asp Arg Arg Tyr Ala Asp Leu Thr Glu Asp Gln Leu Pro Ser Cys Glu 165 170 175 Ser Leu Lys Asp Thr Ile Ala Arg Ala Leu Pro Phe Trp Asn Glu Glu 180 185 190 Ile Val Pro Gln Ile Lys Glu Gly Lys Arg Val Leu Ile Ala Ala His 195 200 205 Gly Asn Ser Leu Arg Gly Ile Val Lys His Leu Glu Gly Leu Ser Glu 210 215 220 Glu Ala Ile Met Glu Leu Asn Leu Pro Thr Gly Ile Pro Ile Val Tyr 225 230 235 240 Glu Leu Asp Lys Asn Leu Lys Pro Ile Lys Pro Met Gln Phe Leu Gly 245 250 255 Met Lys Arg Arg Cys Ala Lys Pro Trp Lys Ala Val Ala Ala Gln Gly 260 265 270 Lys Ala Lys Lys 275 <210> 56 <211> 276 <212> PRT <213> Artificial Sequence <220> <223> GK2-6-PGAM1 fusion protein <400> 56 Tyr Ala Arg Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Ile Ile 1 5 10 15 Arg Ser His Met Leu Glu Met Ala Ala Tyr Lys Leu Val Leu Ile Arg 20 25 30 His Gly Glu Ser Ala Trp Asn Leu Glu Asn Arg Phe Ser Gly Trp Tyr 35 40 45 Asp Ala Asp Leu Ser Pro Ala Gly His Glu Glu Ala Lys Arg Gly Gly 50 55 60 Gln Ala Leu Arg Asp Ala Gly Tyr Glu Phe Asp Ile Cys Phe Thr Ser 65 70 75 80 Val Gln Lys Arg Ala Ile Arg Thr Leu Trp Thr Val Leu Asp Ala Ile 85 90 95 Asp Gln Met Trp Leu Pro Val Val Arg Thr Trp Arg Leu Asn Glu Arg 100 105 110 His Tyr Gly Gly Leu Thr Gly Leu Asn Lys Ala Glu Thr Ala Ala Lys 115 120 125 His Gly Glu Ala Gln Val Lys Ile Trp Arg Arg Ser Tyr Asp Val Pro 130 135 140 Pro Pro Pro Met Glu Pro Asp His Pro Phe Tyr Ser Asn Ile Ser Lys 145 150 155 160 Asp Arg Arg Tyr Ala Asp Leu Thr Glu Asp Gln Leu Pro Ser Cys Glu 165 170 175 Ser Leu Lys Asp Thr Ile Ala Arg Ala Leu Pro Phe Trp Asn Glu Glu 180 185 190 Ile Val Pro Gln Ile Lys Glu Gly Lys Arg Val Leu Ile Ala Ala His 195 200 205 Gly Asn Ser Leu Arg Gly Ile Val Lys His Leu Glu Gly Leu Ser Glu 210 215 220 Glu Ala Ile Met Glu Leu Asn Leu Pro Thr Gly Ile Pro Ile Val Tyr 225 230 235 240 Glu Leu Asp Lys Asn Leu Lys Pro Ile Lys Pro Met Gln Phe Leu Gly 245 250 255 Met Lys Arg Arg Cys Ala Lys Pro Trp Lys Ala Val Ala Ala Gln Gly 260 265 270 Lys Ala Lys Lys 275 <210> 57 <211> 276 <212> PRT <213> Artificial Sequence <220> <223> GK2-7-PGAM1 fusion protein <400> 57 Tyr Ala Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Ile Arg 1 5 10 15 Arg Ser His Met Leu Glu Met Ala Ala Tyr Lys Leu Val Leu Ile Arg 20 25 30 His Gly Glu Ser Ala Trp Asn Leu Glu Asn Arg Phe Ser Gly Trp Tyr 35 40 45 Asp Ala Asp Leu Ser Pro Ala Gly His Glu Glu Ala Lys Arg Gly Gly 50 55 60 Gln Ala Leu Arg Asp Ala Gly Tyr Glu Phe Asp Ile Cys Phe Thr Ser 65 70 75 80 Val Gln Lys Arg Ala Ile Arg Thr Leu Trp Thr Val Leu Asp Ala Ile 85 90 95 Asp Gln Met Trp Leu Pro Val Val Arg Thr Trp Arg Leu Asn Glu Arg 100 105 110 His Tyr Gly Gly Leu Thr Gly Leu Asn Lys Ala Glu Thr Ala Ala Lys 115 120 125 His Gly Glu Ala Gln Val Lys Ile Trp Arg Arg Ser Tyr Asp Val Pro 130 135 140 Pro Pro Pro Met Glu Pro Asp His Pro Phe Tyr Ser Asn Ile Ser Lys 145 150 155 160 Asp Arg Arg Tyr Ala Asp Leu Thr Glu Asp Gln Leu Pro Ser Cys Glu 165 170 175 Ser Leu Lys Asp Thr Ile Ala Arg Ala Leu Pro Phe Trp Asn Glu Glu 180 185 190 Ile Val Pro Gln Ile Lys Glu Gly Lys Arg Val Leu Ile Ala Ala His 195 200 205 Gly Asn Ser Leu Arg Gly Ile Val Lys His Leu Glu Gly Leu Ser Glu 210 215 220 Glu Ala Ile Met Glu Leu Asn Leu Pro Thr Gly Ile Pro Ile Val Tyr 225 230 235 240 Glu Leu Asp Lys Asn Leu Lys Pro Ile Lys Pro Met Gln Phe Leu Gly 245 250 255 Met Lys Arg Arg Cys Ala Lys Pro Trp Lys Ala Val Ala Ala Gln Gly 260 265 270 Lys Ala Lys Lys 275 <210> 58 <211> 276 <212> PRT <213> Artificial Sequence <220> <223> GK2-8-PGAM1 fusion protein <400> 58 Tyr Ala Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Arg Ile 1 5 10 15 Arg Ser His Met Leu Glu Met Ala Ala Tyr Lys Leu Val Leu Ile Arg 20 25 30 His Gly Glu Ser Ala Trp Asn Leu Glu Asn Arg Phe Ser Gly Trp Tyr 35 40 45 Asp Ala Asp Leu Ser Pro Ala Gly His Glu Glu Ala Lys Arg Gly Gly 50 55 60 Gln Ala Leu Arg Asp Ala Gly Tyr Glu Phe Asp Ile Cys Phe Thr Ser 65 70 75 80 Val Gln Lys Arg Ala Ile Arg Thr Leu Trp Thr Val Leu Asp Ala Ile 85 90 95 Asp Gln Met Trp Leu Pro Val Val Arg Thr Trp Arg Leu Asn Glu Arg 100 105 110 His Tyr Gly Gly Leu Thr Gly Leu Asn Lys Ala Glu Thr Ala Ala Lys 115 120 125 His Gly Glu Ala Gln Val Lys Ile Trp Arg Arg Ser Tyr Asp Val Pro 130 135 140 Pro Pro Pro Met Glu Pro Asp His Pro Phe Tyr Ser Asn Ile Ser Lys 145 150 155 160 Asp Arg Arg Tyr Ala Asp Leu Thr Glu Asp Gln Leu Pro Ser Cys Glu 165 170 175 Ser Leu Lys Asp Thr Ile Ala Arg Ala Leu Pro Phe Trp Asn Glu Glu 180 185 190 Ile Val Pro Gln Ile Lys Glu Gly Lys Arg Val Leu Ile Ala Ala His 195 200 205 Gly Asn Ser Leu Arg Gly Ile Val Lys His Leu Glu Gly Leu Ser Glu 210 215 220 Glu Ala Ile Met Glu Leu Asn Leu Pro Thr Gly Ile Pro Ile Val Tyr 225 230 235 240 Glu Leu Asp Lys Asn Leu Lys Pro Ile Lys Pro Met Gln Phe Leu Gly 245 250 255 Met Lys Arg Arg Cys Ala Lys Pro Trp Lys Ala Val Ala Ala Gln Gly 260 265 270 Lys Ala Lys Lys 275 <210> 59 <211> 277 <212> PRT <213> Artificial Sequence <220> <223> GK2-9-PGAM1 fusion protein <400> 59 Arg Tyr Ala Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Ile 1 5 10 15 Ile Arg Ser His Met Leu Glu Met Ala Ala Tyr Lys Leu Val Leu Ile 20 25 30 Arg His Gly Glu Ser Ala Trp Asn Leu Glu Asn Arg Phe Ser Gly Trp 35 40 45 Tyr Asp Ala Asp Leu Ser Pro Ala Gly His Glu Glu Ala Lys Arg Gly 50 55 60 Gly Gln Ala Leu Arg Asp Ala Gly Tyr Glu Phe Asp Ile Cys Phe Thr 65 70 75 80 Ser Val Gln Lys Arg Ala Ile Arg Thr Leu Trp Thr Val Leu Asp Ala 85 90 95 Ile Asp Gln Met Trp Leu Pro Val Val Arg Thr Trp Arg Leu Asn Glu 100 105 110 Arg His Tyr Gly Gly Leu Thr Gly Leu Asn Lys Ala Glu Thr Ala Ala 115 120 125 Lys His Gly Glu Ala Gln Val Lys Ile Trp Arg Arg Ser Tyr Asp Val 130 135 140 Pro Pro Pro Pro Met Glu Pro Asp His Pro Phe Tyr Ser Asn Ile Ser 145 150 155 160 Lys Asp Arg Arg Tyr Ala Asp Leu Thr Glu Asp Gln Leu Pro Ser Cys 165 170 175 Glu Ser Leu Lys Asp Thr Ile Ala Arg Ala Leu Pro Phe Trp Asn Glu 180 185 190 Glu Ile Val Pro Gln Ile Lys Glu Gly Lys Arg Val Leu Ile Ala Ala 195 200 205 His Gly Asn Ser Leu Arg Gly Ile Val Lys His Leu Glu Gly Leu Ser 210 215 220 Glu Glu Ala Ile Met Glu Leu Asn Leu Pro Thr Gly Ile Pro Ile Val 225 230 235 240 Tyr Glu Leu Asp Lys Asn Leu Lys Pro Ile Lys Pro Met Gln Phe Leu 245 250 255 Gly Met Lys Arg Arg Cys Ala Lys Pro Trp Lys Ala Val Ala Ala Gln 260 265 270 Gly Lys Ala Lys Lys 275 <210> 60 <211> 278 <212> PRT <213> Artificial Sequence <220> <223> GK2-10-PGAM1 fusion protein <400> 60 Arg Arg Tyr Ala Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys 1 5 10 15 Ile Ile Arg Ser His Met Leu Glu Met Ala Ala Tyr Lys Leu Val Leu 20 25 30 Ile Arg His Gly Glu Ser Ala Trp Asn Leu Glu Asn Arg Phe Ser Gly 35 40 45 Trp Tyr Asp Ala Asp Leu Ser Pro Ala Gly His Glu Glu Ala Lys Arg 50 55 60 Gly Gly Gln Ala Leu Arg Asp Ala Gly Tyr Glu Phe Asp Ile Cys Phe 65 70 75 80 Thr Ser Val Gln Lys Arg Ala Ile Arg Thr Leu Trp Thr Val Leu Asp 85 90 95 Ala Ile Asp Gln Met Trp Leu Pro Val Val Arg Thr Trp Arg Leu Asn 100 105 110 Glu Arg His Tyr Gly Gly Leu Thr Gly Leu Asn Lys Ala Glu Thr Ala 115 120 125 Ala Lys His Gly Glu Ala Gln Val Lys Ile Trp Arg Arg Ser Tyr Asp 130 135 140 Val Pro Pro Pro Pro Met Glu Pro Asp His Pro Phe Tyr Ser Asn Ile 145 150 155 160 Ser Lys Asp Arg Arg Tyr Ala Asp Leu Thr Glu Asp Gln Leu Pro Ser 165 170 175 Cys Glu Ser Leu Lys Asp Thr Ile Ala Arg Ala Leu Pro Phe Trp Asn 180 185 190 Glu Glu Ile Val Pro Gln Ile Lys Glu Gly Lys Arg Val Leu Ile Ala 195 200 205 Ala His Gly Asn Ser Leu Arg Gly Ile Val Lys His Leu Glu Gly Leu 210 215 220 Ser Glu Glu Ala Ile Met Glu Leu Asn Leu Pro Thr Gly Ile Pro Ile 225 230 235 240 Val Tyr Glu Leu Asp Lys Asn Leu Lys Pro Ile Lys Pro Met Gln Phe 245 250 255 Leu Gly Met Lys Arg Arg Cys Ala Lys Pro Trp Lys Ala Val Ala Ala 260 265 270 Gln Gly Lys Ala Lys Lys 275 <210> 61 <211> 279 <212> PRT <213> Artificial Sequence <220> <223> GK2-11-PGAM1 fusion protein <400> 61 Arg Arg Arg Tyr Ala Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg 1 5 10 15 Lys Ile Ile Arg Ser His Met Leu Glu Met Ala Ala Tyr Lys Leu Val 20 25 30 Leu Ile Arg His Gly Glu Ser Ala Trp Asn Leu Glu Asn Arg Phe Ser 35 40 45 Gly Trp Tyr Asp Ala Asp Leu Ser Pro Ala Gly His Glu Glu Ala Lys 50 55 60 Arg Gly Gly Gln Ala Leu Arg Asp Ala Gly Tyr Glu Phe Asp Ile Cys 65 70 75 80 Phe Thr Ser Val Gln Lys Arg Ala Ile Arg Thr Leu Trp Thr Val Leu 85 90 95 Asp Ala Ile Asp Gln Met Trp Leu Pro Val Val Arg Thr Trp Arg Leu 100 105 110 Asn Glu Arg His Tyr Gly Gly Leu Thr Gly Leu Asn Lys Ala Glu Thr 115 120 125 Ala Ala Lys His Gly Glu Ala Gln Val Lys Ile Trp Arg Arg Ser Tyr 130 135 140 Asp Val Pro Pro Pro Pro Met Glu Pro Asp His Pro Phe Tyr Ser Asn 145 150 155 160 Ile Ser Lys Asp Arg Arg Tyr Ala Asp Leu Thr Glu Asp Gln Leu Pro 165 170 175 Ser Cys Glu Ser Leu Lys Asp Thr Ile Ala Arg Ala Leu Pro Phe Trp 180 185 190 Asn Glu Glu Ile Val Pro Gln Ile Lys Glu Gly Lys Arg Val Leu Ile 195 200 205 Ala Ala His Gly Asn Ser Leu Arg Gly Ile Val Lys His Leu Glu Gly 210 215 220 Leu Ser Glu Glu Ala Ile Met Glu Leu Asn Leu Pro Thr Gly Ile Pro 225 230 235 240 Ile Val Tyr Glu Leu Asp Lys Asn Leu Lys Pro Ile Lys Pro Met Gln 245 250 255 Phe Leu Gly Met Lys Arg Arg Cys Ala Lys Pro Trp Lys Ala Val Ala 260 265 270 Ala Gln Gly Lys Ala Lys Lys 275 <210> 62 <211> 277 <212> PRT <213> Artificial Sequence <220> <223> GK2-12-PGAM1 fusion protein <400> 62 Tyr Ala Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Ile Ile 1 5 10 15 Arg Ser Arg His Met Leu Glu Met Ala Ala Tyr Lys Leu Val Leu Ile 20 25 30 Arg His Gly Glu Ser Ala Trp Asn Leu Glu Asn Arg Phe Ser Gly Trp 35 40 45 Tyr Asp Ala Asp Leu Ser Pro Ala Gly His Glu Glu Ala Lys Arg Gly 50 55 60 Gly Gln Ala Leu Arg Asp Ala Gly Tyr Glu Phe Asp Ile Cys Phe Thr 65 70 75 80 Ser Val Gln Lys Arg Ala Ile Arg Thr Leu Trp Thr Val Leu Asp Ala 85 90 95 Ile Asp Gln Met Trp Leu Pro Val Val Arg Thr Trp Arg Leu Asn Glu 100 105 110 Arg His Tyr Gly Gly Leu Thr Gly Leu Asn Lys Ala Glu Thr Ala Ala 115 120 125 Lys His Gly Glu Ala Gln Val Lys Ile Trp Arg Arg Ser Tyr Asp Val 130 135 140 Pro Pro Pro Pro Met Glu Pro Asp His Pro Phe Tyr Ser Asn Ile Ser 145 150 155 160 Lys Asp Arg Arg Tyr Ala Asp Leu Thr Glu Asp Gln Leu Pro Ser Cys 165 170 175 Glu Ser Leu Lys Asp Thr Ile Ala Arg Ala Leu Pro Phe Trp Asn Glu 180 185 190 Glu Ile Val Pro Gln Ile Lys Glu Gly Lys Arg Val Leu Ile Ala Ala 195 200 205 His Gly Asn Ser Leu Arg Gly Ile Val Lys His Leu Glu Gly Leu Ser 210 215 220 Glu Glu Ala Ile Met Glu Leu Asn Leu Pro Thr Gly Ile Pro Ile Val 225 230 235 240 Tyr Glu Leu Asp Lys Asn Leu Lys Pro Ile Lys Pro Met Gln Phe Leu 245 250 255 Gly Met Lys Arg Arg Cys Ala Lys Pro Trp Lys Ala Val Ala Ala Gln 260 265 270 Gly Lys Ala Lys Lys 275 <210> 63 <211> 278 <212> PRT <213> Artificial Sequence <220> <223> GK2-13-PGAM1 fusion protein <400> 63 Tyr Ala Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Ile Ile 1 5 10 15 Arg Ser Arg Arg His Met Leu Glu Met Ala Ala Tyr Lys Leu Val Leu 20 25 30 Ile Arg His Gly Glu Ser Ala Trp Asn Leu Glu Asn Arg Phe Ser Gly 35 40 45 Trp Tyr Asp Ala Asp Leu Ser Pro Ala Gly His Glu Glu Ala Lys Arg 50 55 60 Gly Gly Gln Ala Leu Arg Asp Ala Gly Tyr Glu Phe Asp Ile Cys Phe 65 70 75 80 Thr Ser Val Gln Lys Arg Ala Ile Arg Thr Leu Trp Thr Val Leu Asp 85 90 95 Ala Ile Asp Gln Met Trp Leu Pro Val Val Arg Thr Trp Arg Leu Asn 100 105 110 Glu Arg His Tyr Gly Gly Leu Thr Gly Leu Asn Lys Ala Glu Thr Ala 115 120 125 Ala Lys His Gly Glu Ala Gln Val Lys Ile Trp Arg Arg Ser Tyr Asp 130 135 140 Val Pro Pro Pro Pro Met Glu Pro Asp His Pro Phe Tyr Ser Asn Ile 145 150 155 160 Ser Lys Asp Arg Arg Tyr Ala Asp Leu Thr Glu Asp Gln Leu Pro Ser 165 170 175 Cys Glu Ser Leu Lys Asp Thr Ile Ala Arg Ala Leu Pro Phe Trp Asn 180 185 190 Glu Glu Ile Val Pro Gln Ile Lys Glu Gly Lys Arg Val Leu Ile Ala 195 200 205 Ala His Gly Asn Ser Leu Arg Gly Ile Val Lys His Leu Glu Gly Leu 210 215 220 Ser Glu Glu Ala Ile Met Glu Leu Asn Leu Pro Thr Gly Ile Pro Ile 225 230 235 240 Val Tyr Glu Leu Asp Lys Asn Leu Lys Pro Ile Lys Pro Met Gln Phe 245 250 255 Leu Gly Met Lys Arg Arg Cys Ala Lys Pro Trp Lys Ala Val Ala Ala 260 265 270 Gln Gly Lys Ala Lys Lys 275 <210> 64 <211> 279 <212> PRT <213> Artificial Sequence <220> <223> GK2-14-PGAM1 fusion protein <400> 64 Tyr Ala Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Ile Ile 1 5 10 15 Arg Ser Arg Arg Arg His Met Leu Glu Met Ala Ala Tyr Lys Leu Val 20 25 30 Leu Ile Arg His Gly Glu Ser Ala Trp Asn Leu Glu Asn Arg Phe Ser 35 40 45 Gly Trp Tyr Asp Ala Asp Leu Ser Pro Ala Gly His Glu Glu Ala Lys 50 55 60 Arg Gly Gly Gln Ala Leu Arg Asp Ala Gly Tyr Glu Phe Asp Ile Cys 65 70 75 80 Phe Thr Ser Val Gln Lys Arg Ala Ile Arg Thr Leu Trp Thr Val Leu 85 90 95 Asp Ala Ile Asp Gln Met Trp Leu Pro Val Val Arg Thr Trp Arg Leu 100 105 110 Asn Glu Arg His Tyr Gly Gly Leu Thr Gly Leu Asn Lys Ala Glu Thr 115 120 125 Ala Ala Lys His Gly Glu Ala Gln Val Lys Ile Trp Arg Arg Ser Tyr 130 135 140 Asp Val Pro Pro Pro Pro Met Glu Pro Asp His Pro Phe Tyr Ser Asn 145 150 155 160 Ile Ser Lys Asp Arg Arg Tyr Ala Asp Leu Thr Glu Asp Gln Leu Pro 165 170 175 Ser Cys Glu Ser Leu Lys Asp Thr Ile Ala Arg Ala Leu Pro Phe Trp 180 185 190 Asn Glu Glu Ile Val Pro Gln Ile Lys Glu Gly Lys Arg Val Leu Ile 195 200 205 Ala Ala His Gly Asn Ser Leu Arg Gly Ile Val Lys His Leu Glu Gly 210 215 220 Leu Ser Glu Glu Ala Ile Met Glu Leu Asn Leu Pro Thr Gly Ile Pro 225 230 235 240 Ile Val Tyr Glu Leu Asp Lys Asn Leu Lys Pro Ile Lys Pro Met Gln 245 250 255 Phe Leu Gly Met Lys Arg Arg Cys Ala Lys Pro Trp Lys Ala Val Ala 260 265 270 Ala Gln Gly Lys Ala Lys Lys 275 <210> 65 <211> 285 <212> PRT <213> Artificial Sequence <220> <223> GK2-15-PGAM1 fusion protein <400> 65 Arg Lys Lys Arg Arg Gln Arg Arg Arg Tyr Ala Tyr Arg Ile Arg Glu 1 5 10 15 Phe Arg Gln Thr Phe Arg Lys Ile Ile Arg Ser His Met Leu Glu Met 20 25 30 Ala Ala Tyr Lys Leu Val Leu Ile Arg His Gly Glu Ser Ala Trp Asn 35 40 45 Leu Glu Asn Arg Phe Ser Gly Trp Tyr Asp Ala Asp Leu Ser Pro Ala 50 55 60 Gly His Glu Glu Ala Lys Arg Gly Gly Gln Ala Leu Arg Asp Ala Gly 65 70 75 80 Tyr Glu Phe Asp Ile Cys Phe Thr Ser Val Gln Lys Arg Ala Ile Arg 85 90 95 Thr Leu Trp Thr Val Leu Asp Ala Ile Asp Gln Met Trp Leu Pro Val 100 105 110 Val Arg Thr Trp Arg Leu Asn Glu Arg His Tyr Gly Gly Leu Thr Gly 115 120 125 Leu Asn Lys Ala Glu Thr Ala Ala Lys His Gly Glu Ala Gln Val Lys 130 135 140 Ile Trp Arg Arg Ser Tyr Asp Val Pro Pro Pro Pro Met Glu Pro Asp 145 150 155 160 His Pro Phe Tyr Ser Asn Ile Ser Lys Asp Arg Arg Tyr Ala Asp Leu 165 170 175 Thr Glu Asp Gln Leu Pro Ser Cys Glu Ser Leu Lys Asp Thr Ile Ala 180 185 190 Arg Ala Leu Pro Phe Trp Asn Glu Glu Ile Val Pro Gln Ile Lys Glu 195 200 205 Gly Lys Arg Val Leu Ile Ala Ala His Gly Asn Ser Leu Arg Gly Ile 210 215 220 Val Lys His Leu Glu Gly Leu Ser Glu Glu Ala Ile Met Glu Leu Asn 225 230 235 240 Leu Pro Thr Gly Ile Pro Ile Val Tyr Glu Leu Asp Lys Asn Leu Lys 245 250 255 Pro Ile Lys Pro Met Gln Phe Leu Gly Met Lys Arg Arg Cys Ala Lys 260 265 270 Pro Trp Lys Ala Val Ala Ala Gln Gly Lys Ala Lys Lys 275 280 285 <210> 66 <211> 281 <212> PRT <213> Artificial Sequence <220> <223> GK2-16-PGAM1 fusion protein <400> 66 Arg Lys Lys Arg Arg Gln Arg Arg Arg Ile Arg Glu Phe Arg Gln Thr 1 5 10 15 Phe Arg Lys Ile Ile Arg Ser His Met Leu Glu Met Ala Ala Tyr Lys 20 25 30 Leu Val Leu Ile Arg His Gly Glu Ser Ala Trp Asn Leu Glu Asn Arg 35 40 45 Phe Ser Gly Trp Tyr Asp Ala Asp Leu Ser Pro Ala Gly His Glu Glu 50 55 60 Ala Lys Arg Gly Gly Gln Ala Leu Arg Asp Ala Gly Tyr Glu Phe Asp 65 70 75 80 Ile Cys Phe Thr Ser Val Gln Lys Arg Ala Ile Arg Thr Leu Trp Thr 85 90 95 Val Leu Asp Ala Ile Asp Gln Met Trp Leu Pro Val Val Arg Thr Trp 100 105 110 Arg Leu Asn Glu Arg His Tyr Gly Gly Leu Thr Gly Leu Asn Lys Ala 115 120 125 Glu Thr Ala Ala Lys His Gly Glu Ala Gln Val Lys Ile Trp Arg Arg 130 135 140 Ser Tyr Asp Val Pro Pro Pro Pro Met Glu Pro Asp His Pro Phe Tyr 145 150 155 160 Ser Asn Ile Ser Lys Asp Arg Arg Tyr Ala Asp Leu Thr Glu Asp Gln 165 170 175 Leu Pro Ser Cys Glu Ser Leu Lys Asp Thr Ile Ala Arg Ala Leu Pro 180 185 190 Phe Trp Asn Glu Glu Ile Val Pro Gln Ile Lys Glu Gly Lys Arg Val 195 200 205 Leu Ile Ala Ala His Gly Asn Ser Leu Arg Gly Ile Val Lys His Leu 210 215 220 Glu Gly Leu Ser Glu Glu Ala Ile Met Glu Leu Asn Leu Pro Thr Gly 225 230 235 240 Ile Pro Ile Val Tyr Glu Leu Asp Lys Asn Leu Lys Pro Ile Lys Pro 245 250 255 Met Gln Phe Leu Gly Met Lys Arg Arg Cys Ala Lys Pro Trp Lys Ala 260 265 270 Val Ala Ala Gln Gly Lys Ala Lys Lys 275 280 <210> 67 <211> 279 <212> PRT <213> Artificial Sequence <220> <223> GK2-17-PGAM1 fusion protein <400> 67 Arg Lys Lys Arg Arg Gln Arg Arg Arg Glu Phe Arg Gln Thr Phe Arg 1 5 10 15 Lys Ile Ile Arg Ser His Met Leu Glu Met Ala Ala Tyr Lys Leu Val 20 25 30 Leu Ile Arg His Gly Glu Ser Ala Trp Asn Leu Glu Asn Arg Phe Ser 35 40 45 Gly Trp Tyr Asp Ala Asp Leu Ser Pro Ala Gly His Glu Glu Ala Lys 50 55 60 Arg Gly Gly Gln Ala Leu Arg Asp Ala Gly Tyr Glu Phe Asp Ile Cys 65 70 75 80 Phe Thr Ser Val Gln Lys Arg Ala Ile Arg Thr Leu Trp Thr Val Leu 85 90 95 Asp Ala Ile Asp Gln Met Trp Leu Pro Val Val Arg Thr Trp Arg Leu 100 105 110 Asn Glu Arg His Tyr Gly Gly Leu Thr Gly Leu Asn Lys Ala Glu Thr 115 120 125 Ala Ala Lys His Gly Glu Ala Gln Val Lys Ile Trp Arg Arg Ser Tyr 130 135 140 Asp Val Pro Pro Pro Pro Met Glu Pro Asp His Pro Phe Tyr Ser Asn 145 150 155 160 Ile Ser Lys Asp Arg Arg Tyr Ala Asp Leu Thr Glu Asp Gln Leu Pro 165 170 175 Ser Cys Glu Ser Leu Lys Asp Thr Ile Ala Arg Ala Leu Pro Phe Trp 180 185 190 Asn Glu Glu Ile Val Pro Gln Ile Lys Glu Gly Lys Arg Val Leu Ile 195 200 205 Ala Ala His Gly Asn Ser Leu Arg Gly Ile Val Lys His Leu Glu Gly 210 215 220 Leu Ser Glu Glu Ala Ile Met Glu Leu Asn Leu Pro Thr Gly Ile Pro 225 230 235 240 Ile Val Tyr Glu Leu Asp Lys Asn Leu Lys Pro Ile Lys Pro Met Gln 245 250 255 Phe Leu Gly Met Lys Arg Arg Cys Ala Lys Pro Trp Lys Ala Val Ala 260 265 270 Ala Gln Gly Lys Ala Lys Lys 275 <210> 68 <211> 283 <212> PRT <213> Artificial Sequence <220> <223> GK2-18-PGAM1 fusion protein <400> 68 Tyr Ala Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Ile Ile 1 5 10 15 Arg Lys Lys Arg Arg Gln Arg Arg Arg His Met Leu Glu Met Ala Ala 20 25 30 Tyr Lys Leu Val Leu Ile Arg His Gly Glu Ser Ala Trp Asn Leu Glu 35 40 45 Asn Arg Phe Ser Gly Trp Tyr Asp Ala Asp Leu Ser Pro Ala Gly His 50 55 60 Glu Glu Ala Lys Arg Gly Gly Gln Ala Leu Arg Asp Ala Gly Tyr Glu 65 70 75 80 Phe Asp Ile Cys Phe Thr Ser Val Gln Lys Arg Ala Ile Arg Thr Leu 85 90 95 Trp Thr Val Leu Asp Ala Ile Asp Gln Met Trp Leu Pro Val Val Arg 100 105 110 Thr Trp Arg Leu Asn Glu Arg His Tyr Gly Gly Leu Thr Gly Leu Asn 115 120 125 Lys Ala Glu Thr Ala Ala Lys His Gly Glu Ala Gln Val Lys Ile Trp 130 135 140 Arg Arg Ser Tyr Asp Val Pro Pro Pro Pro Met Glu Pro Asp His Pro 145 150 155 160 Phe Tyr Ser Asn Ile Ser Lys Asp Arg Arg Tyr Ala Asp Leu Thr Glu 165 170 175 Asp Gln Leu Pro Ser Cys Glu Ser Leu Lys Asp Thr Ile Ala Arg Ala 180 185 190 Leu Pro Phe Trp Asn Glu Glu Ile Val Pro Gln Ile Lys Glu Gly Lys 195 200 205 Arg Val Leu Ile Ala Ala His Gly Asn Ser Leu Arg Gly Ile Val Lys 210 215 220 His Leu Glu Gly Leu Ser Glu Glu Ala Ile Met Glu Leu Asn Leu Pro 225 230 235 240 Thr Gly Ile Pro Ile Val Tyr Glu Leu Asp Lys Asn Leu Lys Pro Ile 245 250 255 Lys Pro Met Gln Phe Leu Gly Met Lys Arg Arg Cys Ala Lys Pro Trp 260 265 270 Lys Ala Val Ala Ala Gln Gly Lys Ala Lys Lys 275 280 <210> 69 <211> 279 <212> PRT <213> Artificial Sequence <220> <223> GK2-19-PGAM1 fusion protein <400> 69 Tyr Ala Tyr Arg Ile Arg Glu Phe Arg Gln Thr Phe Arg Lys Lys Arg 1 5 10 15 Arg Gln Arg Arg Arg His Met Leu Glu Met Ala Ala Tyr Lys Leu Val 20 25 30 Leu Ile Arg His Gly Glu Ser Ala Trp Asn Leu Glu Asn Arg Phe Ser 35 40 45 Gly Trp Tyr Asp Ala Asp Leu Ser Pro Ala Gly His Glu Glu Ala Lys 50 55 60 Arg Gly Gly Gln Ala Leu Arg Asp Ala Gly Tyr Glu Phe Asp Ile Cys 65 70 75 80 Phe Thr Ser Val Gln Lys Arg Ala Ile Arg Thr Leu Trp Thr Val Leu 85 90 95 Asp Ala Ile Asp Gln Met Trp Leu Pro Val Val Arg Thr Trp Arg Leu 100 105 110 Asn Glu Arg His Tyr Gly Gly Leu Thr Gly Leu Asn Lys Ala Glu Thr 115 120 125 Ala Ala Lys His Gly Glu Ala Gln Val Lys Ile Trp Arg Arg Ser Tyr 130 135 140 Asp Val Pro Pro Pro Pro Met Glu Pro Asp His Pro Phe Tyr Ser Asn 145 150 155 160 Ile Ser Lys Asp Arg Arg Tyr Ala Asp Leu Thr Glu Asp Gln Leu Pro 165 170 175 Ser Cys Glu Ser Leu Lys Asp Thr Ile Ala Arg Ala Leu Pro Phe Trp 180 185 190 Asn Glu Glu Ile Val Pro Gln Ile Lys Glu Gly Lys Arg Val Leu Ile 195 200 205 Ala Ala His Gly Asn Ser Leu Arg Gly Ile Val Lys His Leu Glu Gly 210 215 220 Leu Ser Glu Glu Ala Ile Met Glu Leu Asn Leu Pro Thr Gly Ile Pro 225 230 235 240 Ile Val Tyr Glu Leu Asp Lys Asn Leu Lys Pro Ile Lys Pro Met Gln 245 250 255 Phe Leu Gly Met Lys Arg Arg Cys Ala Lys Pro Trp Lys Ala Val Ala 260 265 270 Ala Gln Gly Lys Ala Lys Lys 275 <210> 70 <211> 289 <212> PRT <213> Artificial Sequence <220> <223> GK2-20-PGAM1 fusion protein <400> 70 Lys Glu Thr Trp Trp Glu Thr Trp Trp Thr Glu Trp Ser Gln Pro Lys 1 5 10 15 Lys Lys Arg Lys Val Arg Gln Thr Phe Arg Lys Ile Ile Arg Ser His 20 25 30 Met Leu Glu Met Ala Ala Tyr Lys Leu Val Leu Ile Arg His Gly Glu 35 40 45 Ser Ala Trp Asn Leu Glu Asn Arg Phe Ser Gly Trp Tyr Asp Ala Asp 50 55 60 Leu Ser Pro Ala Gly His Glu Glu Ala Lys Arg Gly Gly Gln Ala Leu 65 70 75 80 Arg Asp Ala Gly Tyr Glu Phe Asp Ile Cys Phe Thr Ser Val Gln Lys 85 90 95 Arg Ala Ile Arg Thr Leu Trp Thr Val Leu Asp Ala Ile Asp Gln Met 100 105 110 Trp Leu Pro Val Val Arg Thr Trp Arg Leu Asn Glu Arg His Tyr Gly 115 120 125 Gly Leu Thr Gly Leu Asn Lys Ala Glu Thr Ala Ala Lys His Gly Glu 130 135 140 Ala Gln Val Lys Ile Trp Arg Arg Ser Tyr Asp Val Pro Pro Pro Pro 145 150 155 160 Met Glu Pro Asp His Pro Phe Tyr Ser Asn Ile Ser Lys Asp Arg Arg 165 170 175 Tyr Ala Asp Leu Thr Glu Asp Gln Leu Pro Ser Cys Glu Ser Leu Lys 180 185 190 Asp Thr Ile Ala Arg Ala Leu Pro Phe Trp Asn Glu Glu Ile Val Pro 195 200 205 Gln Ile Lys Glu Gly Lys Arg Val Leu Ile Ala Ala His Gly Asn Ser 210 215 220 Leu Arg Gly Ile Val Lys His Leu Glu Gly Leu Ser Glu Glu Ala Ile 225 230 235 240 Met Glu Leu Asn Leu Pro Thr Gly Ile Pro Ile Val Tyr Glu Leu Asp 245 250 255 Lys Asn Leu Lys Pro Ile Lys Pro Met Gln Phe Leu Gly Met Lys Arg 260 265 270 Arg Cys Ala Lys Pro Trp Lys Ala Val Ala Ala Gln Gly Lys Ala Lys 275 280 285 Lys <210> 71 <211> 287 <212> PRT <213> Artificial Sequence <220> <223> GK2-21-PGAM1 fusion protein <400> 71 Tyr Ala Tyr Arg Ile Arg Glu Phe Lys Glu Thr Trp Trp Glu Thr Trp 1 5 10 15 Trp Thr Glu Trp Ser Gln Pro Lys Lys Lys Arg Lys Val His Met Leu 20 25 30 Glu Met Ala Ala Tyr Lys Leu Val Leu Ile Arg His Gly Glu Ser Ala 35 40 45 Trp Asn Leu Glu Asn Arg Phe Ser Gly Trp Tyr Asp Ala Asp Leu Ser 50 55 60 Pro Ala Gly His Glu Glu Ala Lys Arg Gly Gly Gln Ala Leu Arg Asp 65 70 75 80 Ala Gly Tyr Glu Phe Asp Ile Cys Phe Thr Ser Val Gln Lys Arg Ala 85 90 95 Ile Arg Thr Leu Trp Thr Val Leu Asp Ala Ile Asp Gln Met Trp Leu 100 105 110 Pro Val Val Arg Thr Trp Arg Leu Asn Glu Arg His Tyr Gly Gly Leu 115 120 125 Thr Gly Leu Asn Lys Ala Glu Thr Ala Ala Lys His Gly Glu Ala Gln 130 135 140 Val Lys Ile Trp Arg Arg Ser Tyr Asp Val Pro Pro Pro Pro Met Glu 145 150 155 160 Pro Asp His Pro Phe Tyr Ser Asn Ile Ser Lys Asp Arg Arg Tyr Ala 165 170 175 Asp Leu Thr Glu Asp Gln Leu Pro Ser Cys Glu Ser Leu Lys Asp Thr 180 185 190 Ile Ala Arg Ala Leu Pro Phe Trp Asn Glu Glu Ile Val Pro Gln Ile 195 200 205 Lys Glu Gly Lys Arg Val Leu Ile Ala Ala His Gly Asn Ser Leu Arg 210 215 220 Gly Ile Val Lys His Leu Glu Gly Leu Ser Glu Glu Ala Ile Met Glu 225 230 235 240 Leu Asn Leu Pro Thr Gly Ile Pro Ile Val Tyr Glu Leu Asp Lys Asn 245 250 255 Leu Lys Pro Ile Lys Pro Met Gln Phe Leu Gly Met Lys Arg Arg Cys 260 265 270 Ala Lys Pro Trp Lys Ala Val Ala Ala Gln Gly Lys Ala Lys Lys 275 280 285 <210> 72 <211> 831 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for GK2-PGAM1 fusion protein <400> 72 tacgcctacc gtatccgcga gttccgccag accttccgca agatcattcg cagccatatg 60 ctcgagatgg ccgcctacaa actggtgctg atccggcacg gcgagagcgc atggaacctg 120 gagaaccgct tcagcggctg gtacgacgcc gacctgagcc cggcgggcca cgaggaggcg 180 aagcgcggcg ggcaggcgct acgagatgct ggctatgagt ttgacatctg cttcacctca 240 gtgcagaaga gagcgatccg gaccctctgg acagtgctag atgccattga tcagatgtgg 300 ctgccagtgg tgaggacttg gcgcctcaat gagcggcact atgggggtct aaccggtctc 360 aataaagcag aaactgctgc aaagcatggt gaggcccagg tgaagatctg gaggcgctcc 420 tatgatgtcc caccacctcc gatggagccc gaccatcctt tctacagcaa catcagtaag 480 gatcgcaggt atgcagacct cacagaagat cagctaccct cctgtgagag tctgaaggat 540 actattgcca gagctctgcc cttctggaat gaagaaatag ttccccagat caaggagggg 600 aaacgtgtac tgattgcagc ccatggcaac agcctccggg gcattgtcaa gcatctggag 660 ggtctctctg aagaggctat catggagctg aacctgccga ctggtattcc cattgtctat 720 gaattggaca agaacttgaa gcctatcaag cccatgcagt ttctggggat gaagagacgg 780 tgcgcaaagc catggaaagc tgtggctgcc cagggcaagg ccaagaagtg a 831 <210> 73 <211> 831 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for GK2-0-PGAM1 fusion protein <400> 73 cgcgcctacc gtatccgcga gttccgccag accttccgca agatcattcg cagccatatg 60 ctcgagatgg ccgcctacaa actggtgctg atccggcacg gcgagagcgc atggaacctg 120 gagaaccgct tcagcggctg gtacgacgcc gacctgagcc cggcgggcca cgaggaggcg 180 aagcgcggcg ggcaggcgct acgagatgct ggctatgagt ttgacatctg cttcacctca 240 gtgcagaaga gagcgatccg gaccctctgg acagtgctag atgccattga tcagatgtgg 300 ctgccagtgg tgaggacttg gcgcctcaat gagcggcact atgggggtct aaccggtctc 360 aataaagcag aaactgctgc aaagcatggt gaggcccagg tgaagatctg gaggcgctcc 420 tatgatgtcc caccacctcc gatggagccc gaccatcctt tctacagcaa catcagtaag 480 gatcgcaggt atgcagacct cacagaagat cagctaccct cctgtgagag tctgaaggat 540 actattgcca gagctctgcc cttctggaat gaagaaatag ttccccagat caaggagggg 600 aaacgtgtac tgattgcagc ccatggcaac agcctccggg gcattgtcaa gcatctggag 660 ggtctctctg aagaggctat catggagctg aacctgccga ctggtattcc cattgtctat 720 gaattggaca agaacttgaa gcctatcaag cccatgcagt ttctggggat gaagagacgg 780 tgcgcaaagc catggaaagc tgtggctgcc cagggcaagg ccaagaagtg a 831 <210> 74 <211> 831 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for GK2-1-PGAM1 fusion protein <400> 74 cgccgttacc gtatccgcga gttccgccag accttccgca agatcattcg cagccatatg 60 ctcgagatgg ccgcctacaa actggtgctg atccggcacg gcgagagcgc atggaacctg 120 gagaaccgct tcagcggctg gtacgacgcc gacctgagcc cggcgggcca cgaggaggcg 180 aagcgcggcg ggcaggcgct acgagatgct ggctatgagt ttgacatctg cttcacctca 240 gtgcagaaga gagcgatccg gaccctctgg acagtgctag atgccattga tcagatgtgg 300 ctgccagtgg tgaggacttg gcgcctcaat gagcggcact atgggggtct aaccggtctc 360 aataaagcag aaactgctgc aaagcatggt gaggcccagg tgaagatctg gaggcgctcc 420 tatgatgtcc caccacctcc gatggagccc gaccatcctt tctacagcaa catcagtaag 480 gatcgcaggt atgcagacct cacagaagat cagctaccct cctgtgagag tctgaaggat 540 actattgcca gagctctgcc cttctggaat gaagaaatag ttccccagat caaggagggg 600 aaacgtgtac tgattgcagc ccatggcaac agcctccggg gcattgtcaa gcatctggag 660 ggtctctctg aagaggctat catggagctg aacctgccga ctggtattcc cattgtctat 720 gaattggaca agaacttgaa gcctatcaag cccatgcagt ttctggggat gaagagacgg 780 tgcgcaaagc catggaaagc tgtggctgcc cagggcaagg ccaagaagtg a 831 <210> 75 <211> 831 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for GK2-2-PGAM1 fusion protein <400> 75 cgccgtcgcc gtatccgcga gttccgccag accttccgca agatcattcg cagccatatg 60 ctcgagatgg ccgcctacaa actggtgctg atccggcacg gcgagagcgc atggaacctg 120 gagaaccgct tcagcggctg gtacgacgcc gacctgagcc cggcgggcca cgaggaggcg 180 aagcgcggcg ggcaggcgct acgagatgct ggctatgagt ttgacatctg cttcacctca 240 gtgcagaaga gagcgatccg gaccctctgg acagtgctag atgccattga tcagatgtgg 300 ctgccagtgg tgaggacttg gcgcctcaat gagcggcact atgggggtct aaccggtctc 360 aataaagcag aaactgctgc aaagcatggt gaggcccagg tgaagatctg gaggcgctcc 420 tatgatgtcc caccacctcc gatggagccc gaccatcctt tctacagcaa catcagtaag 480 gatcgcaggt atgcagacct cacagaagat cagctaccct cctgtgagag tctgaaggat 540 actattgcca gagctctgcc cttctggaat gaagaaatag ttccccagat caaggagggg 600 aaacgtgtac tgattgcagc ccatggcaac agcctccggg gcattgtcaa gcatctggag 660 ggtctctctg aagaggctat catggagctg aacctgccga ctggtattcc cattgtctat 720 gaattggaca agaacttgaa gcctatcaag cccatgcagt ttctggggat gaagagacgg 780 tgcgcaaagc catggaaagc tgtggctgcc cagggcaagg ccaagaagtg a 831 <210> 76 <211> 831 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for GK2-3-PGAM1 fusion protein <400> 76 tacgcctacc gtatccgcga gttccgccag accttccgca agatcattcg ccgccatatg 60 ctcgagatgg ccgcctacaa actggtgctg atccggcacg gcgagagcgc atggaacctg 120 gagaaccgct tcagcggctg gtacgacgcc gacctgagcc cggcgggcca cgaggaggcg 180 aagcgcggcg ggcaggcgct acgagatgct ggctatgagt ttgacatctg cttcacctca 240 gtgcagaaga gagcgatccg gaccctctgg acagtgctag atgccattga tcagatgtgg 300 ctgccagtgg tgaggacttg gcgcctcaat gagcggcact atgggggtct aaccggtctc 360 aataaagcag aaactgctgc aaagcatggt gaggcccagg tgaagatctg gaggcgctcc 420 tatgatgtcc caccacctcc gatggagccc gaccatcctt tctacagcaa catcagtaag 480 gatcgcaggt atgcagacct cacagaagat cagctaccct cctgtgagag tctgaaggat 540 actattgcca gagctctgcc cttctggaat gaagaaatag ttccccagat caaggagggg 600 aaacgtgtac tgattgcagc ccatggcaac agcctccggg gcattgtcaa gcatctggag 660 ggtctctctg aagaggctat catggagctg aacctgccga ctggtattcc cattgtctat 720 gaattggaca agaacttgaa gcctatcaag cccatgcagt ttctggggat gaagagacgg 780 tgcgcaaagc catggaaagc tgtggctgcc cagggcaagg ccaagaagtg a 831 <210> 77 <211> 831 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for GK2-4-PGAM1 fusion protein <400> 77 tacgcctacc gtatccgcga gttccgccag accttccgca agatccgtcg ccgccatatg 60 ctcgagatgg ccgcctacaa actggtgctg atccggcacg gcgagagcgc atggaacctg 120 gagaaccgct tcagcggctg gtacgacgcc gacctgagcc cggcgggcca cgaggaggcg 180 aagcgcggcg ggcaggcgct acgagatgct ggctatgagt ttgacatctg cttcacctca 240 gtgcagaaga gagcgatccg gaccctctgg acagtgctag atgccattga tcagatgtgg 300 ctgccagtgg tgaggacttg gcgcctcaat gagcggcact atgggggtct aaccggtctc 360 aataaagcag aaactgctgc aaagcatggt gaggcccagg tgaagatctg gaggcgctcc 420 tatgatgtcc caccacctcc gatggagccc gaccatcctt tctacagcaa catcagtaag 480 gatcgcaggt atgcagacct cacagaagat cagctaccct cctgtgagag tctgaaggat 540 actattgcca gagctctgcc cttctggaat gaagaaatag ttccccagat caaggagggg 600 aaacgtgtac tgattgcagc ccatggcaac agcctccggg gcattgtcaa gcatctggag 660 ggtctctctg aagaggctat catggagctg aacctgccga ctggtattcc cattgtctat 720 gaattggaca agaacttgaa gcctatcaag cccatgcagt ttctggggat gaagagacgg 780 tgcgcaaagc catggaaagc tgtggctgcc cagggcaagg ccaagaagtg a 831 <210> 78 <211> 831 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for GK2-5-PGAM1 fusion protein <400> 78 tacgcctacc gtatccgcga gttccgccag accttccgca agcgccgtcg ccgccatatg 60 ctcgagatgg ccgcctacaa actggtgctg atccggcacg gcgagagcgc atggaacctg 120 gagaaccgct tcagcggctg gtacgacgcc gacctgagcc cggcgggcca cgaggaggcg 180 aagcgcggcg ggcaggcgct acgagatgct ggctatgagt ttgacatctg cttcacctca 240 gtgcagaaga gagcgatccg gaccctctgg acagtgctag atgccattga tcagatgtgg 300 ctgccagtgg tgaggacttg gcgcctcaat gagcggcact atgggggtct aaccggtctc 360 aataaagcag aaactgctgc aaagcatggt gaggcccagg tgaagatctg gaggcgctcc 420 tatgatgtcc caccacctcc gatggagccc gaccatcctt tctacagcaa catcagtaag 480 gatcgcaggt atgcagacct cacagaagat cagctaccct cctgtgagag tctgaaggat 540 actattgcca gagctctgcc cttctggaat gaagaaatag ttccccagat caaggagggg 600 aaacgtgtac tgattgcagc ccatggcaac agcctccggg gcattgtcaa gcatctggag 660 ggtctctctg aagaggctat catggagctg aacctgccga ctggtattcc cattgtctat 720 gaattggaca agaacttgaa gcctatcaag cccatgcagt ttctggggat gaagagacgg 780 tgcgcaaagc catggaaagc tgtggctgcc cagggcaagg ccaagaagtg a 831 <210> 79 <211> 831 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for GK2-6-PGAM1 fusion protein <400> 79 tacgcccgcc gtatccgcga gttccgccag accttccgca agatcattcg cagccatatg 60 ctcgagatgg ccgcctacaa actggtgctg atccggcacg gcgagagcgc atggaacctg 120 gagaaccgct tcagcggctg gtacgacgcc gacctgagcc cggcgggcca cgaggaggcg 180 aagcgcggcg ggcaggcgct acgagatgct ggctatgagt ttgacatctg cttcacctca 240 gtgcagaaga gagcgatccg gaccctctgg acagtgctag atgccattga tcagatgtgg 300 ctgccagtgg tgaggacttg gcgcctcaat gagcggcact atgggggtct aaccggtctc 360 aataaagcag aaactgctgc aaagcatggt gaggcccagg tgaagatctg gaggcgctcc 420 tatgatgtcc caccacctcc gatggagccc gaccatcctt tctacagcaa catcagtaag 480 gatcgcaggt atgcagacct cacagaagat cagctaccct cctgtgagag tctgaaggat 540 actattgcca gagctctgcc cttctggaat gaagaaatag ttccccagat caaggagggg 600 aaacgtgtac tgattgcagc ccatggcaac agcctccggg gcattgtcaa gcatctggag 660 ggtctctctg aagaggctat catggagctg aacctgccga ctggtattcc cattgtctat 720 gaattggaca agaacttgaa gcctatcaag cccatgcagt ttctggggat gaagagacgg 780 tgcgcaaagc catggaaagc tgtggctgcc cagggcaagg ccaagaagtg a 831 <210> 80 <211> 831 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for GK2-7-PGAM1 fusion protein <400> 80 tacgcctacc gtatccgcga gttccgccag accttccgca agatccgccg cagccatatg 60 ctcgagatgg ccgcctacaa actggtgctg atccggcacg gcgagagcgc atggaacctg 120 gagaaccgct tcagcggctg gtacgacgcc gacctgagcc cggcgggcca cgaggaggcg 180 aagcgcggcg ggcaggcgct acgagatgct ggctatgagt ttgacatctg cttcacctca 240 gtgcagaaga gagcgatccg gaccctctgg acagtgctag atgccattga tcagatgtgg 300 ctgccagtgg tgaggacttg gcgcctcaat gagcggcact atgggggtct aaccggtctc 360 aataaagcag aaactgctgc aaagcatggt gaggcccagg tgaagatctg gaggcgctcc 420 tatgatgtcc caccacctcc gatggagccc gaccatcctt tctacagcaa catcagtaag 480 gatcgcaggt atgcagacct cacagaagat cagctaccct cctgtgagag tctgaaggat 540 actattgcca gagctctgcc cttctggaat gaagaaatag ttccccagat caaggagggg 600 aaacgtgtac tgattgcagc ccatggcaac agcctccggg gcattgtcaa gcatctggag 660 ggtctctctg aagaggctat catggagctg aacctgccga ctggtattcc cattgtctat 720 gaattggaca agaacttgaa gcctatcaag cccatgcagt ttctggggat gaagagacgg 780 tgcgcaaagc catggaaagc tgtggctgcc cagggcaagg ccaagaagtg a 831 <210> 81 <211> 831 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for GK2-8-PGAM1 fusion protein <400> 81 tacgcctacc gtatccgcga gttccgccag accttccgca agcgcattcg cagccatatg 60 ctcgagatgg ccgcctacaa actggtgctg atccggcacg gcgagagcgc atggaacctg 120 gagaaccgct tcagcggctg gtacgacgcc gacctgagcc cggcgggcca cgaggaggcg 180 aagcgcggcg ggcaggcgct acgagatgct ggctatgagt ttgacatctg cttcacctca 240 gtgcagaaga gagcgatccg gaccctctgg acagtgctag atgccattga tcagatgtgg 300 ctgccagtgg tgaggacttg gcgcctcaat gagcggcact atgggggtct aaccggtctc 360 aataaagcag aaactgctgc aaagcatggt gaggcccagg tgaagatctg gaggcgctcc 420 tatgatgtcc caccacctcc gatggagccc gaccatcctt tctacagcaa catcagtaag 480 gatcgcaggt atgcagacct cacagaagat cagctaccct cctgtgagag tctgaaggat 540 actattgcca gagctctgcc cttctggaat gaagaaatag ttccccagat caaggagggg 600 aaacgtgtac tgattgcagc ccatggcaac agcctccggg gcattgtcaa gcatctggag 660 ggtctctctg aagaggctat catggagctg aacctgccga ctggtattcc cattgtctat 720 gaattggaca agaacttgaa gcctatcaag cccatgcagt ttctggggat gaagagacgg 780 tgcgcaaagc catggaaagc tgtggctgcc cagggcaagg ccaagaagtg a 831 <210> 82 <211> 834 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for GK2-9-PGAM1 fusion protein <400> 82 cgctacgcct accgtatccg cgagttccgc cagaccttcc gcaagatcat tcgcagccat 60 atgctcgaga tggccgccta caaactggtg ctgatccggc acggcgagag cgcatggaac 120 ctggagaacc gcttcagcgg ctggtacgac gccgacctga gcccggcggg ccacgaggag 180 gcgaagcgcg gcgggcaggc gctacgagat gctggctatg agtttgacat ctgcttcacc 240 tcagtgcaga agagagcgat ccggaccctc tggacagtgc tagatgccat tgatcagatg 300 tggctgccag tggtgaggac ttggcgcctc aatgagcggc actatggggg tctaaccggt 360 ctcaataaag cagaaactgc tgcaaagcat ggtgaggccc aggtgaagat ctggaggcgc 420 tcctatgatg tcccaccacc tccgatggag cccgaccatc ctttctacag caacatcagt 480 aaggatcgca ggtatgcaga cctcacagaa gatcagctac cctcctgtga gagtctgaag 540 gatactattg ccagagctct gcccttctgg aatgaagaaa tagttcccca gatcaaggag 600 gggaaacgtg tactgattgc agcccatggc aacagcctcc ggggcattgt caagcatctg 660 gagggtctct ctgaagaggc tatcatggag ctgaacctgc cgactggtat tcccattgtc 720 tatgaattgg acaagaactt gaagcctatc aagcccatgc agtttctggg gatgaagaga 780 cggtgcgcaa agccatggaa agctgtggct gcccagggca aggccaagaa gtga 834 <210> 83 <211> 837 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for GK2-10-PGAM1 fusion protein <400> 83 cgccgttacg cctaccgtat ccgcgagttc cgccagacct tccgcaagat cattcgcagc 60 catatgctcg agatggccgc ctacaaactg gtgctgatcc ggcacggcga gagcgcatgg 120 aacctggaga accgcttcag cggctggtac gacgccgacc tgagcccggc gggccacgag 180 gaggcgaagc gcggcgggca ggcgctacga gatgctggct atgagtttga catctgcttc 240 acctcagtgc agaagagagc gatccggacc ctctggacag tgctagatgc cattgatcag 300 atgtggctgc cagtggtgag gacttggcgc ctcaatgagc ggcactatgg gggtctaacc 360 ggtctcaata aagcagaaac tgctgcaaag catggtgagg cccaggtgaa gatctggagg 420 cgctcctatg atgtcccacc acctccgatg gagcccgacc atcctttcta cagcaacatc 480 agtaaggatc gcaggtatgc agacctcaca gaagatcagc taccctcctg tgagagtctg 540 aaggatacta ttgccagagc tctgcccttc tggaatgaag aaatagttcc ccagatcaag 600 gaggggaaac gtgtactgat tgcagcccat ggcaacagcc tccggggcat tgtcaagcat 660 ctggagggtc tctctgaaga ggctatcatg gagctgaacc tgccgactgg tattcccatt 720 gtctatgaat tggacaagaa cttgaagcct atcaagccca tgcagtttct ggggatgaag 780 agacggtgcg caaagccatg gaaagctgtg gctgcccagg gcaaggccaa gaagtga 837 <210> 84 <211> 840 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for GK2-11-PGAM1 fusion protein <400> 84 cgccgtcgct acgcctaccg tatccgcgag ttccgccaga ccttccgcaa gatcattcgc 60 agccatatgc tcgagatggc cgcctacaaa ctggtgctga tccggcacgg cgagagcgca 120 tggaacctgg agaaccgctt cagcggctgg tacgacgccg acctgagccc ggcgggccac 180 gaggaggcga agcgcggcgg gcaggcgcta cgagatgctg gctatgagtt tgacatctgc 240 ttcacctcag tgcagaagag agcgatccgg accctctgga cagtgctaga tgccattgat 300 cagatgtggc tgccagtggt gaggacttgg cgcctcaatg agcggcacta tgggggtcta 360 accggtctca ataaagcaga aactgctgca aagcatggtg aggcccaggt gaagatctgg 420 aggcgctcct atgatgtccc accacctccg atggagcccg accatccttt ctacagcaac 480 atcagtaagg atcgcaggta tgcagacctc acagaagatc agctaccctc ctgtgagagt 540 ctgaaggata ctattgccag agctctgccc ttctggaatg aagaaatagt tccccagatc 600 aaggagggga aacgtgtact gattgcagcc catggcaaca gcctccgggg cattgtcaag 660 catctggagg gtctctctga agaggctatc atggagctga acctgccgac tggtattccc 720 attgtctatg aattggacaa gaacttgaag cctatcaagc ccatgcagtt tctggggatg 780 aagagacggt gcgcaaagcc atggaaagct gtggctgccc agggcaaggc caagaagtga 840 840 <210> 85 <211> 834 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for GK2-12-PGAM1 fusion protein <400> 85 tacgcctacc gtatccgcga gttccgccag accttccgca agatcattcg cagccgccat 60 atgctcgaga tggccgccta caaactggtg ctgatccggc acggcgagag cgcatggaac 120 ctggagaacc gcttcagcgg ctggtacgac gccgacctga gcccggcggg ccacgaggag 180 gcgaagcgcg gcgggcaggc gctacgagat gctggctatg agtttgacat ctgcttcacc 240 tcagtgcaga agagagcgat ccggaccctc tggacagtgc tagatgccat tgatcagatg 300 tggctgccag tggtgaggac ttggcgcctc aatgagcggc actatggggg tctaaccggt 360 ctcaataaag cagaaactgc tgcaaagcat ggtgaggccc aggtgaagat ctggaggcgc 420 tcctatgatg tcccaccacc tccgatggag cccgaccatc ctttctacag caacatcagt 480 aaggatcgca ggtatgcaga cctcacagaa gatcagctac cctcctgtga gagtctgaag 540 gatactattg ccagagctct gcccttctgg aatgaagaaa tagttcccca gatcaaggag 600 gggaaacgtg tactgattgc agcccatggc aacagcctcc ggggcattgt caagcatctg 660 gagggtctct ctgaagaggc tatcatggag ctgaacctgc cgactggtat tcccattgtc 720 tatgaattgg acaagaactt gaagcctatc aagcccatgc agtttctggg gatgaagaga 780 cggtgcgcaa agccatggaa agctgtggct gcccagggca aggccaagaa gtga 834 <210> 86 <211> 837 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for GK2-13-PGAM1 fusion protein <400> 86 tacgcctacc gtatccgcga gttccgccag accttccgca agatcattcg cagccgccgt 60 catatgctcg agatggccgc ctacaaactg gtgctgatcc ggcacggcga gagcgcatgg 120 aacctggaga accgcttcag cggctggtac gacgccgacc tgagcccggc gggccacgag 180 gaggcgaagc gcggcgggca ggcgctacga gatgctggct atgagtttga catctgcttc 240 acctcagtgc agaagagagc gatccggacc ctctggacag tgctagatgc cattgatcag 300 atgtggctgc cagtggtgag gacttggcgc ctcaatgagc ggcactatgg gggtctaacc 360 ggtctcaata aagcagaaac tgctgcaaag catggtgagg cccaggtgaa gatctggagg 420 cgctcctatg atgtcccacc acctccgatg gagcccgacc atcctttcta cagcaacatc 480 agtaaggatc gcaggtatgc agacctcaca gaagatcagc taccctcctg tgagagtctg 540 aaggatacta ttgccagagc tctgcccttc tggaatgaag aaatagttcc ccagatcaag 600 gaggggaaac gtgtactgat tgcagcccat ggcaacagcc tccggggcat tgtcaagcat 660 ctggagggtc tctctgaaga ggctatcatg gagctgaacc tgccgactgg tattcccatt 720 gtctatgaat tggacaagaa cttgaagcct atcaagccca tgcagtttct ggggatgaag 780 agacggtgcg caaagccatg gaaagctgtg gctgcccagg gcaaggccaa gaagtga 837 <210> 87 <211> 840 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for GK2-14-PGAM1 fusion protein <400> 87 tacgcctacc gtatccgcga gttccgccag accttccgca agatcattcg cagccgccgt 60 cgccatatgc tcgagatggc cgcctacaaa ctggtgctga tccggcacgg cgagagcgca 120 tggaacctgg agaaccgctt cagcggctgg tacgacgccg acctgagccc ggcgggccac 180 gaggaggcga agcgcggcgg gcaggcgcta cgagatgctg gctatgagtt tgacatctgc 240 ttcacctcag tgcagaagag agcgatccgg accctctgga cagtgctaga tgccattgat 300 cagatgtggc tgccagtggt gaggacttgg cgcctcaatg agcggcacta tgggggtcta 360 accggtctca ataaagcaga aactgctgca aagcatggtg aggcccaggt gaagatctgg 420 aggcgctcct atgatgtccc accacctccg atggagcccg accatccttt ctacagcaac 480 atcagtaagg atcgcaggta tgcagacctc acagaagatc agctaccctc ctgtgagagt 540 ctgaaggata ctattgccag agctctgccc ttctggaatg aagaaatagt tccccagatc 600 aaggagggga aacgtgtact gattgcagcc catggcaaca gcctccgggg cattgtcaag 660 catctggagg gtctctctga agaggctatc atggagctga acctgccgac tggtattccc 720 attgtctatg aattggacaa gaacttgaag cctatcaagc ccatgcagtt tctggggatg 780 aagagacggt gcgcaaagcc atggaaagct gtggctgccc agggcaaggc caagaagtga 840 840 <210> 88 <211> 858 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for GK2-15-PGAM1 fusion protein <400> 88 aggaagaagc ggagacagcg acgaagatac gcctaccgta tccgcgagtt ccgccagacc 60 ttccgcaaga tcattcgcag ccatatgctc gagatggccg cctacaaact ggtgctgatc 120 cggcacggcg agagcgcatg gaacctggag aaccgcttca gcggctggta cgacgccgac 180 ctgagcccgg cgggccacga ggaggcgaag cgcggcgggc aggcgctacg agatgctggc 240 tatgagtttg acatctgctt cacctcagtg cagaagagag cgatccggac cctctggaca 300 gtgctagatg ccattgatca gatgtggctg ccagtggtga ggacttggcg cctcaatgag 360 cggcactatg ggggtctaac cggtctcaat aaagcagaaa ctgctgcaaa gcatggtgag 420 gcccaggtga agatctggag gcgctcctat gatgtcccac cacctccgat ggagcccgac 480 catcctttct acagcaacat cagtaaggat cgcaggtatg cagacctcac agaagatcag 540 ctaccctcct gtgagagtct gaaggatact attgccagag ctctgccctt ctggaatgaa 600 gaaatagttc cccagatcaa ggaggggaaa cgtgtactga ttgcagccca tggcaacagc 660 ctccggggca ttgtcaagca tctggagggt ctctctgaag aggctatcat ggagctgaac 720 ctgccgactg gtattcccat tgtctatgaa ttggacaaga acttgaagcc tatcaagccc 780 atgcagtttc tggggatgaa gagacggtgc gcaaagccat ggaaagctgt ggctgcccag 840 ggcaaggcca agaagtga 858 <210> 89 <211> 846 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for GK2-16-PGAM1 fusion protein <400> 89 aggaagaagc ggagacagcg acgaagaatc cgcgagttcc gccagacctt ccgcaagatc 60 attcgcagcc atatgctcga gatggccgcc tacaaactgg tgctgatccg gcacggcgag 120 agcgcatgga acctggagaa ccgcttcagc ggctggtacg acgccgacct gagcccggcg 180 ggccacgagg aggcgaagcg cggcgggcag gcgctacgag atgctggcta tgagtttgac 240 atctgcttca cctcagtgca gaagagagcg atccggaccc tctggacagt gctagatgcc 300 attgatcaga tgtggctgcc agtggtgagg acttggcgcc tcaatgagcg gcactatggg 360 ggtctaaccg gtctcaataa agcagaaact gctgcaaagc atggtgaggc ccaggtgaag 420 atctggaggc gctcctatga tgtcccacca cctccgatgg agcccgacca tcctttctac 480 agcaacatca gtaaggatcg caggtatgca gacctcacag aagatcagct accctcctgt 540 gagagtctga aggatactat tgccagagct ctgcccttct ggaatgaaga aatagttccc 600 cagatcaagg aggggaaacg tgtactgatt gcagcccatg gcaacagcct ccggggcatt 660 gtcaagcatc tggagggtct ctctgaagag gctatcatgg agctgaacct gccgactggt 720 attcccattg tctatgaatt ggacaagaac ttgaagccta tcaagcccat gcagtttctg 780 gggatgaaga gacggtgcgc aaagccatgg aaagctgtgg ctgcccaggg caaggccaag 840 aagtga 846 <210> 90 <211> 840 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for GK2-17-PGAM1 fusion protein <400> 90 aggaagaagc ggagacagcg acgaagagag ttccgccaga ccttccgcaa gatcattcgc 60 agccatatgc tcgagatggc cgcctacaaa ctggtgctga tccggcacgg cgagagcgca 120 tggaacctgg agaaccgctt cagcggctgg tacgacgccg acctgagccc ggcgggccac 180 gaggaggcga agcgcggcgg gcaggcgcta cgagatgctg gctatgagtt tgacatctgc 240 ttcacctcag tgcagaagag agcgatccgg accctctgga cagtgctaga tgccattgat 300 cagatgtggc tgccagtggt gaggacttgg cgcctcaatg agcggcacta tgggggtcta 360 accggtctca ataaagcaga aactgctgca aagcatggtg aggcccaggt gaagatctgg 420 aggcgctcct atgatgtccc accacctccg atggagcccg accatccttt ctacagcaac 480 atcagtaagg atcgcaggta tgcagacctc acagaagatc agctaccctc ctgtgagagt 540 ctgaaggata ctattgccag agctctgccc ttctggaatg aagaaatagt tccccagatc 600 aaggagggga aacgtgtact gattgcagcc catggcaaca gcctccgggg cattgtcaag 660 catctggagg gtctctctga agaggctatc atggagctga acctgccgac tggtattccc 720 attgtctatg aattggacaa gaacttgaag cctatcaagc ccatgcagtt tctggggatg 780 aagagacggt gcgcaaagcc atggaaagct gtggctgccc agggcaaggc caagaagtga 840 840 <210> 91 <211> 852 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for GK2-18-PGAM1 fusion protein <400> 91 tacgcctacc gtatccgcga gttccgccag accttccgca agatcattcg caagaagcgg 60 agacagcgac gaagacatat gctcgagatg gccgcctaca aactggtgct gatccggcac 120 ggcgagagcg catggaacct ggagaaccgc ttcagcggct ggtacgacgc cgacctgagc 180 ccggcgggcc acgaggaggc gaagcgcggc gggcaggcgc tacgagatgc tggctatgag 240 tttgacatct gcttcacctc agtgcagaag agagcgatcc ggaccctctg gacagtgcta 300 gatgccattg atcagatgtg gctgccagtg gtgaggactt ggcgcctcaa tgagcggcac 360 tatgggggtc taaccggtct caataaagca gaaactgctg caaagcatgg tgaggcccag 420 gtgaagatct ggaggcgctc ctatgatgtc ccaccacctc cgatggagcc cgaccatcct 480 ttctacagca acatcagtaa ggatcgcagg tatgcagacc tcacagaaga tcagctaccc 540 tcctgtgaga gtctgaagga tactattgcc agagctctgc ccttctggaa tgaagaaata 600 gttccccaga tcaaggaggg gaaacgtgta ctgattgcag cccatggcaa cagcctccgg 660 ggcattgtca agcatctgga gggtctctct gaagaggcta tcatggagct gaacctgccg 720 actggtattc ccattgtcta tgaattggac aagaacttga agcctatcaa gcccatgcag 780 tttctgggga tgaagagacg gtgcgcaaag ccatggaaag ctgtggctgc ccagggcaag 840 gccaagaagt ga 852 <210> 92 <211> 840 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for GK2-19-PGAM1 fusion protein <400> 92 tacgcctacc gtatccgcga gttccgccag accttccgca agaagcggag acagcgacga 60 agacatatgc tcgagatggc cgcctacaaa ctggtgctga tccggcacgg cgagagcgca 120 tggaacctgg agaaccgctt cagcggctgg tacgacgccg acctgagccc ggcgggccac 180 gaggaggcga agcgcggcgg gcaggcgcta cgagatgctg gctatgagtt tgacatctgc 240 ttcacctcag tgcagaagag agcgatccgg accctctgga cagtgctaga tgccattgat 300 cagatgtggc tgccagtggt gaggacttgg cgcctcaatg agcggcacta tgggggtcta 360 accggtctca ataaagcaga aactgctgca aagcatggtg aggcccaggt gaagatctgg 420 aggcgctcct atgatgtccc accacctccg atggagcccg accatccttt ctacagcaac 480 atcagtaagg atcgcaggta tgcagacctc acagaagatc agctaccctc ctgtgagagt 540 ctgaaggata ctattgccag agctctgccc ttctggaatg aagaaatagt tccccagatc 600 aaggagggga aacgtgtact gattgcagcc catggcaaca gcctccgggg cattgtcaag 660 catctggagg gtctctctga agaggctatc atggagctga acctgccgac tggtattccc 720 attgtctatg aattggacaa gaacttgaag cctatcaagc ccatgcagtt tctggggatg 780 aagagacggt gcgcaaagcc atggaaagct gtggctgccc agggcaaggc caagaagtga 840 840 <210> 93 <211> 870 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for GK2-20-PGAM1 fusion protein <400> 93 aaagaaacct ggtgggaaac ctggtggacc gaatggtctc agccgaaaaa aaaacgtaaa 60 gtgcgccaga ccttccgcaa gatcattcgc agccatatgc tcgagatggc cgcctacaaa 120 ctggtgctga tccggcacgg cgagagcgca tggaacctgg agaaccgctt cagcggctgg 180 tacgacgccg acctgagccc ggcgggccac gaggaggcga agcgcggcgg gcaggcgcta 240 cgagatgctg gctatgagtt tgacatctgc ttcacctcag tgcagaagag agcgatccgg 300 accctctgga cagtgctaga tgccattgat cagatgtggc tgccagtggt gaggacttgg 360 cgcctcaatg agcggcacta tgggggtcta accggtctca ataaagcaga aactgctgca 420 aagcatggtg aggcccaggt gaagatctgg aggcgctcct atgatgtccc accacctccg 480 atggagcccg accatccttt ctacagcaac atcagtaagg atcgcaggta tgcagacctc 540 acagaagatc agctaccctc ctgtgagagt ctgaaggata ctattgccag agctctgccc 600 ttctggaatg aagaaatagt tccccagatc aaggagggga aacgtgtact gattgcagcc 660 catggcaaca gcctccgggg cattgtcaag catctggagg gtctctctga agaggctatc 720 atggagctga acctgccgac tggtattccc attgtctatg aattggacaa gaacttgaag 780 cctatcaagc ccatgcagtt tctggggatg aagagacggt gcgcaaagcc atggaaagct 840 gtggctgccc agggcaaggc caagaagtga 870 <210> 94 <211> 864 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for GK2-21-PGAM1 fusion protein <400> 94 tacgcctacc gtatccgcga gttcaaagaa acctggtggg aaacctggtg gaccgaatgg 60 tctcagccga aaaaaaaacg taaagtgcat atgctcgaga tggccgccta caaactggtg 120 ctgatccggc acggcgagag cgcatggaac ctggagaacc gcttcagcgg ctggtacgac 180 gccgacctga gcccggcggg ccacgaggag gcgaagcgcg gcgggcaggc gctacgagat 240 gctggctatg agtttgacat ctgcttcacc tcagtgcaga agagagcgat ccggaccctc 300 tggacagtgc tagatgccat tgatcagatg tggctgccag tggtgaggac ttggcgcctc 360 aatgagcggc actatggggg tctaaccggt ctcaataaag cagaaactgc tgcaaagcat 420 ggtgaggccc aggtgaagat ctggaggcgc tcctatgatg tcccaccacc tccgatggag 480 cccgaccatc ctttctacag caacatcagt aaggatcgca ggtatgcaga cctcacagaa 540 gatcagctac cctcctgtga gagtctgaag gatactattg ccagagctct gcccttctgg 600 aatgaagaaa tagttcccca gatcaaggag gggaaacgtg tactgattgc agcccatggc 660 aacagcctcc ggggcattgt caagcatctg gagggtctct ctgaagaggc tatcatggag 720 ctgaacctgc cgactggtat tcccattgtc tatgaattgg acaagaactt gaagcctatc 780 aagcccatgc agtttctggg gatgaagaga cggtgcgcaa agccatggaa agctgtggct 840 gcccagggca aggccaagaa gtga 864

Claims (12)

As one selected from the following (A) to (D), phosphoglycerate mutase 1 to a cell-penetrating peptide consisting of 16 to 31 amino acids, which binds to cargo molecules and transports cargo molecules into mammalian cells or tissues. This fused cell-penetrating phosphoglycerate mutase 1 fusion protein.
(A) R ₁ R ₂ R ₃ RIREFRQTFRKR ₄ R ₅ R ₆ R ₇ ,
(However, R ₁ is _{one selected} from Y, R, RR, RRR, RRRR, RY, RRY, RRRY, R ₂ is one selected from A and R, R ₃ is one selected from Y and R, and R ₄ is one selected from I and R, R ₅ is one selected from I and R, R ₆ is R, R ₇ is one selected from S, R, RR, RRR, RRRR, SR, SRR, SRRR),
(B) R ₈ REFRQTFRKR ₉ ,
(However, R ₈ is one selected from RKKRRQRRRYAYRI, RKKRRQRRRI, RKKRRQRR and YAYRI, R ₉ is one selected from IIRS, IIRKKRRQRRR and KRRQRRR),
(C) R ₁₀ RQTFRKIIRS
(However, R ₁₀ is 19 or more consecutive amino acids in the KETWWETWWTEWSQPKKKRKV sequence),
(D) YAYRIREFR ₁₀
(However, R ₁₀ is 19 or more consecutive amino acids in the KETWWETWWTEWSQPKKKRKV sequence).
The method according to claim 1,
The cell-penetrating phosphoglycerate mutase 1 fusion protein is a cell-penetrating phosphoglycerate mutase 1 fusion protein, characterized in that one selected from SEQ ID NO: 49 to SEQ ID NO: 71.
The method according to claim 1,
Cell-penetrating phosphoglycerate mutase 1 fusion protein, characterized in that the fusion of the cell-penetrating peptide and phosphoglycerate mutase 1 is by a covalent bond.
delete delete delete delete Recombinant polynucleotide encoding the cell-penetrating phosphoglycerate mutase 1 fusion protein of any one of claims 1 to 3.
The method of claim 8,
A recombinant polynucleotide encoding a cell-penetrating phosphoglycerate mutase 1 fusion protein, characterized in that the oligonucleotide sequence encoding the cell-penetrating peptide is one selected from SEQ ID NO: 72 to SEQ ID NO: 94.
delete Phosphoglycerate mutase 1 fusion protein expression vector comprising a recombinant polynucleotide encoding the cell-penetrating phosphoglycerate mutase 1 fusion protein of claim 8.
delete

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