CN116802197A

CN116802197A - Pharmaceutical composition for preventing or treating tuberculosis

Info

Publication number: CN116802197A
Application number: CN202180092760.8A
Authority: CN
Inventors: 徐裕锡; 朴基硕
Original assignee: Department Of Disease Management; Genexine Inc
Current assignee: Department Of Disease Management; Genexine Inc
Priority date: 2020-12-04
Filing date: 2021-12-03
Publication date: 2023-09-22
Also published as: WO2022119384A1; KR20220079081A

Abstract

The present invention relates to a fusion protein comprising: a fusion polypeptide derived from mycobacterium tuberculosis (Mycoba terium tuberculosis); a polynucleotide encoding the above protein; a recombinant vector comprising the polynucleotide; and a pharmaceutical composition for preventing or treating tuberculosis, comprising the above fusion protein, polynucleotide or recombinant vector.

Description

Pharmaceutical composition for preventing or treating tuberculosis

Technical Field

Background

Tuberculosis is an infectious disease caused by mycobacterium tuberculosis (Mycobacterium Tuberculosis), and when an infectious tuberculosis patient coughs or sneezes, the mycobacterium tuberculosis is discharged into the air, and surrounding people may inhale the droplets to be infected. In general, although not every infected person is a tuberculosis patient, even if the infected person is infected with mycobacterium tuberculosis, the infection is not affected in the normal immune system state, and the infection can also live in normal life. Therefore, there is a need to develop a vaccine for preventing or treating tuberculosis to protect people not infected with tuberculosis or people infected with tuberculosis.

BCG vaccine (bacillus Calmette-Gurin, BCG) is an attenuated live vaccine for weakening bovine type mycobacterium tuberculosis (Mycobacterium bovis), is the most commonly used tuberculosis vaccine worldwide, and can effectively protect children. However, it has a problem that the duration is limited to childhood, and the preventive effect on adult tuberculosis, which is a major form of tuberculosis, is in an unstable state of 0 to 80%. Thus, new tuberculosis vaccines are being developed in countries around the world that can replace or supplement existing tuberculosis vaccines.

The developed novel tuberculosis vaccine comprises: protein subunit vaccines that utilize tuberculosis antigens; expressing a tuberculosis antigen inducing defensive immunity in a DN A vaccine or a viral vector vaccine in a plasmid DNA or a viral vector; recombinant BCG vaccine expressing a tuberculosis antigen inducing defensive immunity on BCG vaccine of tuberculosis antigen; a gene recombinant tuberculosis vaccine for removing pathogenic genes of tuberculosis bacteria, etc. To date, DNA vaccines have been tried more frequently as novel tuberculosis preventive vaccines, and their efficacy has been demonstrated in animal models. In addition, many tuberculosis antigens have been developed using DNA vaccine technology.

DNA vaccines have proven to be very effective in establishing cellular immune responses including cytotoxic T lymphocytes (cytotoxic Tlymphocyte, CTLs) and Th1 responses, which are known to be critical for obtaining effective antibacterial immunity (anti-mycobacteria immunity) in mice and humans (Bonato VL et al, effect Immun,1998, 66:169-175). In addition, efforts are underway to develop antigens because of the importance of selecting appropriate antigens that are critical to the efficacy of the defense against tuberculosis in terms of introducing effective immunological techniques.

For this reason, the present inventors developed a vaccine for preventing or treating tuberculosis using an antigen having a defensive efficacy against tuberculosis, and completed the present invention.

Disclosure of Invention

Technical problem

It is an object of the present invention to provide a fusion protein comprising a fusion polypeptide derived from Mycobacterium tuberculosis.

It is still another object of the present invention to provide a polynucleotide for encoding the above fusion protein.

It is another object of the present invention to provide a recombinant vector comprising the above polynucleotide.

It is still another object of the present invention to provide a pharmaceutical composition for preventing or treating tuberculosis comprising the above fusion protein.

It is still another object of the present invention to provide a pharmaceutical composition for preventing or treating tuberculosis comprising the above polynucleotide.

It is still another object of the present invention to provide a pharmaceutical composition for preventing or treating tuberculosis comprising the above recombinant vector.

It is a further object of the present invention to provide a method for preventing or treating tuberculosis, comprising the step of administering a therapeutically effective amount of the above fusion protein to a subject.

It is a further object of the present invention to provide a method for preventing or treating tuberculosis, comprising the step of administering a therapeutically effective amount of the polynucleotide as described above to a subject.

It is still another object of the present invention to provide a method for preventing or treating tuberculosis, comprising the step of administering a therapeutically effective amount of the recombinant vector as described above to a subject.

It is a further object of the present invention to provide the use of the fusion protein as defined above for the preparation of a medicament for the prevention or treatment of tuberculosis.

It is a further object of the present invention to provide the use of the above polynucleotide for the preparation of a medicament for preventing or treating tuberculosis.

It is a further object of the present invention to provide the use of the recombinant vector described above in the preparation of a medicament for the prevention or treatment of tuberculosis.

Means for solving the problems

In order to achieve the above object, the present invention provides a fusion protein comprising a fusion polypeptide derived from Mycobacterium tuberculosis.

Also, the present invention provides polynucleotides for encoding the above fusion proteins.

The present invention also provides a recombinant vector comprising the polynucleotide.

Also, the present invention provides a pharmaceutical composition for preventing or treating tuberculosis, comprising the above fusion protein.

Also, the present invention provides a pharmaceutical composition for preventing or treating tuberculosis, comprising the above polynucleotide.

And, the present invention provides a pharmaceutical composition for preventing or treating tuberculosis comprising the above recombinant vector.

Also, the present invention provides a method for preventing or treating tuberculosis, comprising the step of administering a therapeutically effective amount of the above fusion protein to a subject.

Also, the present invention provides a method for preventing or treating tuberculosis, comprising the step of administering a therapeutically effective amount of the above polynucleotide to a subject.

Also, the present invention provides a method for preventing or treating tuberculosis, comprising the step of administering a therapeutically effective amount of the recombinant vector described above to a subject.

Furthermore, the present invention provides the use of the above fusion protein in the preparation of a medicament for preventing or treating tuberculosis.

And, the present invention provides the use of the above polynucleotide for the preparation of a medicament for preventing or treating tuberculosis.

Furthermore, the invention provides the use of the recombinant vector in preparing a medicament for preventing or treating tuberculosis.

ADVANTAGEOUS EFFECTS OF INVENTION

The fusion protein provided by the invention has the effects of increasing the specific immune response of a human body to tuberculosis antigens and remarkably reducing the quantity of tubercle bacillus in the lung, and can be effectively used for preventing or treating tuberculosis.

Drawings

FIG. 1 shows a gene construct for preparing a fusion protein.

FIG. 2 shows the results of specific immune responses to tuberculosis antigens as determined by IFN-. Gamma.ELISPOT analysis after administration of DNA vaccine to a mouse animal model.

FIG. 3 shows the results of determining the number of Mycobacterium tuberculosis in the lung and the results of lung histopathology after administration of DNA vaccine to a mouse animal model.

FIG. 4 shows the results of determining the number of Mycobacterium tuberculosis in the lung and the results of lung histopathology after administration of DNA vaccine to a mouse animal model.

FIG. 5 shows the results of determining the number of Mycobacterium tuberculosis in the lung after administration of DNA vaccine to a mouse animal model and reinfection with Mycobacterium tuberculosis.

Detailed Description

Best Mode for Carrying Out The Invention

The present invention provides fusion proteins comprising a fusion polypeptide derived from mycobacterium tuberculosis.

The fusion polypeptide may be selected from the group consisting of Ag85A, TB 10.4.4, mtb32b and PstS3, and preferably may comprise Ag85A, TB 10.4.4, mtb32b and PstS3. In an embodiment of the invention, the Ag85A may consist of the amino acid sequence of seq id No. 1; the above TB10.4 may consist of the amino acid sequence of SEQ ID NO. 2; the Mtb32b may consist of the amino acid sequence of seq id No. 3; the above PstS3 may consist of the amino acid sequence of SEQ ID No. 4.

The fusion polypeptide may further comprise a polypeptide selected from the group consisting of PPE39, glcB and RipA, and preferably may further comprise a polypeptide comprising PPE39, glcB and RipA. In one embodiment of the present invention, the PPE39 may consist of the amino acid sequence of SEQ ID No. 5; the above GlcB may consist of the amino acid sequence of seq id No. 6; the RipA may consist of the amino acid sequence of SEQ ID No. 7.

The fusion polypeptide may be linked by a 2A peptide (2A self-cleaning peptide) or a GS linker, which may preferably comprise (GS) n (in this case, n is an integer from 1 to 10). In one embodiment of the present invention, the GS linker may be composed of the amino acid sequence of SEQ ID NO. 8.

In the present invention, the above fusion protein may further comprise a signal peptide.

The term "signal peptide" or secretion signal sequence "in the present invention refers to a short peptide of about 20 to 30 amino acids in length, which exists at the N-terminus of a newly synthesized protein designated as secretion pathway (secretory pathway).

The signal peptide may be selected from the group consisting of a tissue-type plasminogen activator (tissue plaminoge n activator, tPA) signal peptide, a herpes simplex virus glycoprotein Ds (Herpes Simplex Virus glycoprotein Ds, HSV gDs) signal peptide, and a growth hormone (growth hormo) signal peptide, and preferably, a secretion signal sequence used in higher eukaryotic cells including mammals, and more preferably, a tPA sequence may be used.

In one embodiment of the present invention, the tPA sequence may consist of the amino acid sequence of seq id No. 9. Furthermore, the signal sequence of the present invention may be replaced with a codon having a high expression frequency in the host cell.

In the present invention, the above fusion protein further comprises an immunopotentiating peptide.

The term "immunopotentiating peptide" in the present invention refers to a peptide that increases an immune response by activating cells (e.g., dendritic cells, etc.) involved in the immune response.

The immunopotentiating peptide may be selected from the group consisting of CD40 ligand, FMS-like tyrosine kinase 3 ligand (FMS-like tyrosine kinase-3, flt 3) ligand, flagellin (flagellin) and OX40, preferably, but not limited thereto, flt3 ligand. In one embodiment of the present invention, the Flt3 ligand may consist of the amino acid sequence of SEQ ID NO. 10.

The present invention provides a fusion protein comprising: a signal peptide; immunopotentiating peptides of Flt3 ligands; and fusion polypeptides comprising Ag85A, TB10.4, mtb32b and PstS3.

In one embodiment of the present invention, the fusion protein may consist of the amino acid sequence of SEQ ID NO. 11.

Also, the present invention provides a fusion protein comprising: a signal peptide; immunopotentiating peptides of Flt3 ligands; and fusion polypeptides comprising PPE39, glcB and RipA.

In one embodiment of the present invention, the fusion protein may consist of the amino acid sequence of SEQ ID NO. 12.

The polynucleotides of the invention may be prepared by chemical synthesis or genetic engineering techniques. Chemical synthesis methods are known to those of ordinary skill in the art to which the present invention pertains, and any method may be used. Or by commission of commercial nucleic acid synthesis and purchase by suppliers. In the case of preparation by genetic engineering techniques, for example, it can be prepared by obtaining nucleic acid fragments encoding a fusion polypeptide, a signal peptide and an immunopotentiating peptide derived from Mycobacterium tuberculosis, which have been known in the past, respectively, and ligating these fragments in a manner suitable for the frame. Methods for obtaining such nucleic acid fragments are well known in the art to which the present invention pertains, and one of ordinary skill in the art can readily ligate using appropriate restriction enzymes. In particular embodiments of the invention, methods of preparation using chemical synthesis are disclosed.

The polynucleotide of the present invention may comprise a polynucleotide for encoding a fusion polypeptide comprising Ag85A, TB10.4, mtb32b and PstS3.

The Ag85A may consist of the nucleic acid sequence of SEQ ID NO. 13; the TB10.4 may consist of the nucleic acid sequence of SEQ ID NO. 14; the Mtb32b may be composed of the nucleic acid sequence of SEQ ID NO. 15; the above PstS3 may consist of the nucleic acid sequence of SEQ ID NO. 16. Ag85A, TB 10.4.4, mtb32b and PstS3 can be linked by a GS linker, which can preferably consist of a nucleic acid sequence selected from sequence numbers 20 to 23.

Also, the polynucleotide of the present invention may comprise a polynucleotide encoding a fusion polypeptide comprising PPE39, glcB and RipA.

The PPE39 may consist of the nucleic acid sequence of SEQ ID NO. 17; the above GlcB may consist of the nucleic acid sequence of seq id No. 18; the above RipA may consist of the nucleic acid sequence of SEQ ID NO. 19. PPE39, glcB and RipA may be linked by a GS linker, which may preferably consist of a nucleic acid sequence selected from sequence numbers 20 to 23.

The polynucleotide of the present invention may further comprise a polynucleotide encoding a signal peptide, wherein the signal peptide may be a tPA sequence, and the polynucleotide encoding the tPA sequence may be composed of a nucleic acid sequence of sequence No. 24 or 25.

The polynucleotide of the present invention may further comprise a polynucleotide encoding an immunopotentiating peptide, which may be a Flt3 ligand, and the polynucleotide encoding the Flt3 ligand may be composed of the nucleic acid sequence of SEQ ID NO. 26 or 27.

In one embodiment of the present invention, the polynucleotide may comprise: a polynucleotide for encoding a fusion polypeptide comprising Ag85A, TB10.4, mtb32b, and PstS 3; a polynucleotide encoding a signal peptide; a polynucleotide for encoding an immunopotentiating peptide. Preferably, the polynucleotide may consist of the nucleic acid sequence of SEQ ID NO. 28.

In one embodiment of the present invention, the polynucleotide may comprise: a polynucleotide encoding a fusion polypeptide comprising PPE39, glcB, and RipA; a polynucleotide encoding a signal peptide; polynucleotides encoding immunopotentiating peptides. Preferably, the polynucleotide may consist of the nucleic acid sequence of SEQ ID NO. 29.

The term "vector" according to the present invention refers to a genetic construct comprising an external DNA inserted into the genome encoding a polypeptide. The recombinant expression vector of the present invention is a vector in which a polynucleotide encoding a fusion protein comprising a fusion polypeptide derived from Mycobacterium tuberculosis, a signal peptide and an immunopotentiating peptide is inserted into the genome.

The vector may be a plasmid vector, a viral vector, a cosmid vector, or an artificial chromosome vector, and the viral vector may be a phage vector, an adenovirus vector, a retrovirus vector, or an adeno-associated virus vector.

The recombinant vector of the invention may comprise a nucleic acid capable of encoding the fusion protein of the invention in a form suitable for expression of the nucleic acid encoding the fusion protein in a host cell. That is, the recombinant vectors of the invention comprise one or more regulatory sequences selected according to the host cell used for expression, which are operably linked to the nucleic acid sequence to be expressed.

The term "operably linked" in the present invention refers to a desired nucleotide sequence (e.g., in an in vitro transcription/translation system or host cell) linked to the regulatory sequences described above in a manner that allows for its expression.

The term "regulatory sequence" in the present invention refers to promoters, enhancers and other regulatory elements (e.g., polyadenylation signals). Regulatory sequences include sequences that indicate expression of a desired nucleic acid in a number of host cells, as well as sequences that indicate that the desired nucleic acid is capable of expression only in a particular host cell (e.g., tissue-specific regulatory sequences). It will be appreciated by those of ordinary skill in the art that the design of the expression vector may vary depending on factors such as the choice of host cell to be transformed and the desired level of protein expression. The expression vectors of the invention may express the fusion proteins described above by introduction into a host cell.

In one embodiment of the present invention, the composition may further comprise an immunoadjuvant (adjuvant) comprising interleukin-12 (IL-12), interleukin-21 (IL-21) and macrophage inflammatory protein-1α (MIP-1 a) proteins, preferably the immunoadjuvant may comprise an amino acid sequence of sequence No. 30 to 33. Wherein IL-12p35 consists of the amino acid sequence of SEQ ID NO. 30; IL-12p40 consists of the amino acid sequence of SEQ ID NO. 31; IL-21 consists of the amino acid sequence of SEQ ID NO. 32; MIP-1a (MIP-1 alpha) consists of the amino acid sequence of SEQ ID NO. 33.

The term "immunoadjuvant" according to the present invention refers to a substance added to a vaccine to stimulate the immune system response of an antigen.

Also, the present invention provides a pharmaceutical composition for preventing or treating tuberculosis, comprising the above polynucleotide or the above recombinant vector.

The above composition may further comprise a polynucleotide encoding an immunoadjuvant comprising IL-12, IL-21 and MIP-1a proteins, preferably a polynucleotide encoding an amino acid sequence of SEQ ID Nos. 30 to 33, more preferably a nucleic acid sequence of SEQ ID Nos. 34 to 37.

The pharmaceutical compositions of the present invention may also comprise a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable carrier" in the context of the present invention means that it exhibits non-toxic properties to cells or humans exposed to the above-described compositions. And carriers known in the art to which the present invention pertains, such as buffers, preservatives, painless agents, solubilizers, isotonic agents, stabilizers, bases, excipients, lubricants and the like, may be used without limitation.

The composition of the present invention can be formulated into non-oral preparations such as oral preparations or injections, for example, granules, powders, solutions, tablets, capsules, dry syrups, and the like. Preferably, the composition of the present invention may be in the form of a solution or an injection, etc., but is not limited thereto.

In the pharmaceutical compositions of the present invention, the pharmaceutically effective amount is administered. The term "administration" according to the present invention means that a predetermined substance is introduced into a subject in an appropriate manner, and the administration route of the above-mentioned composition may be by any conventional route as long as it is capable of reaching a desired tissue. Intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, oral administration, topical administration, intranasal administration, intrapulmonary administration, and intrarectal administration may be used, but are not limited thereto.

The term "subject" refers to all animals including humans, such as rats, mice, and livestock, among others. Preferably, it may be a mammal, including a human.

The term "pharmaceutically effective amount" as defined above refers to an amount sufficient to treat a disease without causing side effects in a reasonable benefit/risk ratio applicable to medical treatment, and the effective amount level can be readily determined by one of ordinary skill in the art based on such conditions as sex, age, weight, health, disease type, severity, pharmaceutical activity, sensitivity to drugs, method of administration, time of administration, route and rate of administration, period of treatment, elements combining or containing simultaneous drugs, and other elements well known in the medical arts of the patient. At the time of administration, the administration may be carried out once daily or in several times according to the recommended doses described above.

Also, the present invention provides a method for preventing or treating tuberculosis, comprising the step of administering a therapeutically effective amount of the above polynucleotide or the above recombinant vector to a subject.

Preferably, the above therapeutically effective amount may be variously applied according to the type and extent of the reaction to be achieved, the specific composition of other formulation or the like according to circumstances, the age, weight, general health, sex and diet of the subject, administration time, administration route and secretion rate of the composition, treatment period, drug used together or simultaneously with the specific composition, and the like, and similar factors widely applied in the medical field. Thus, preferably, an effective amount of a composition suitable for the purposes of the present invention can be determined based on the factors described above.

The subject may be applied to any mammal, including not only humans and primates, but also livestock such as cows, pigs, sheep, horses, dogs, cats, and the like.

Furthermore, the present invention provides the use of the above polynucleotide or the above recombinant vector for the preparation of a medicament for preventing or treating tuberculosis.

Embodiments of the invention

Hereinafter, the present invention will be described in more detail with reference to examples. These examples are intended to illustrate the present invention in more detail, but the scope of the present invention is not limited to these examples only.

EXAMPLE 1 preparation of fusion proteins

The fusion protein of the present invention comprises a fusion polypeptide derived from Mycobacterium tuberculosis, a signal peptide and an immunopotentiator peptide (immunity enhancer peptide). The above fusion polypeptide derived from Mycobacterium tuberculosis contains Ag85A, TB 10.4.4, mtb32b and PstS3, and each polypeptide (antigen) is prepared by 2A peptide ligation. And, the tPA peptide in the signal peptide and the Flt3L ligand in the immunopotentiating peptide are linked to the N-terminal of the fusion polypeptide.

The fusion polypeptide derived from Mycobacterium tuberculosis was prepared by including a polypeptide of PPE39, glcB and/or RipA in addition to Ag85A, TB 10.4.10.4, mt b32b and PstS3. Each polypeptide (antigen) was prepared by ligation via a 2A peptide or GS linker, and was also prepared at the N-terminus of the fusion polypeptide as a Flt3L ligand in the tPA peptide and the immunopotentiating peptide in the signal peptide.

Hereinafter, the fusion polypeptide of Ag85A, TB 10.4.4, mtb32B and PstS3 was linked by a 2A peptide, the fusion protein in which HA tag was added to the C-terminus was designated "GX-70", the fusion protein in which Ag85A, TB 10.4.4, mtb32B, PPE39 and PstS3 was linked by a 2A peptide was designated "GX-171A", the fusion protein in which Ag85A, TB 10.4.4, mtb32B, ripA and PstS3 was linked by a 2A peptide was designated "GX-171B", the fusion protein in which Ag85A, TB 10.4.4, mtb32B and PstS3 was linked by a GS linker was designated "GX-170A", and the fusion protein in which PPE39, glcB and RipA were linked by a GS linker was designated "GX-170B" (FIG. 1).

The above candidate fusion proteins were prepared by synthesizing pGX27 (see Korean patent No. 1442254) having the gene sequence of each corresponding fusion protein inserted into a high expression vector.

EXAMPLE 2 preparation of immunoadjuvant

The immunoadjuvant of the present invention comprises IL-12, IL-21 and MIP-1a proteins. The N-terminus of IL-12 was linked to CMV promoter (pCMV), the N-terminus of IL-21 was linked to RSV promoter (pRSV), and the N-terminus of MIP-1a was linked to EF1a promoter (pEF 1 a). The immunoadjuvant comprising IL-12, IL-21 and MIP-1A proteins is hereinafter referred to as "BD-121A" (FIG. 1).

Prepared by synthesizing a cassette for expressing each corresponding fusion protein and inserting it into pGX27 (refer to korean patent No. 1442254) of a high expression vector.

EXAMPLE 3 immunopotentiating efficacy analysis of compositions comprising fusion proteins and immunoadjuvants

The present inventors have conducted experiments to confirm whether or not a composition comprising the fusion protein and an immunoadjuvant has an immunopotentiating effect. Briefly, use is made of(SL VAXiGEN, korea) in vivo electroporation apparatus PBS (dosage form control) or DNA vaccine was administered to thigh muscle of C57BL/6 mice 2 times every 2 weeks. Spleens were removed 2 weeks after final administration and specific immune responses to tuberculosis antigens were determined using IFN-gamma ELISPOT analysis. Information for PBS or DNA vaccine dosing groups are as follows: group 1, pbs (dosage form control); group 2, GX-70 μg; group 3, GX-70 μg+BD-121A8 μg; group 4, GX-171A8 μg; group 5, GX-171A8 μg+BD-121A8 μg; group 6, GX-171B 8 μg; and group 7, GX-171B 8 μg+BD-121A8 μg.

As a result, it was confirmed that tuberculosis specific immune responses were induced in tuberculosis candidate substances GX-70, GX-171A and GX-171B, and in particular, in groups 3, 5 and 7, in which the immunoadjuvant BD-121A was added to GX-70, GX-171A and GX-171B, the tuberculosis specific immune responses were further increased (FIG. 2).

EXAMPLE 4 analysis of the prevention efficacy of tuberculosis

The present inventors conducted experimentsTo confirm whether the composition comprising the fusion protein and the immunoadjuvant has the effect of preventing tuberculosis. Briefly, use is made of(SL VAXiGE N, korea) in vivo electroporation apparatus PBS (dosage form control) or DNA vaccine was administered on thigh muscle of C57BL/6 mice 3 times every 3 weeks. In the positive control BCG group, C57BL/6 mice were administered 1 time at week 3. After 3 weeks of final administration, mycobacterium tuberculosis was released into the air, and after 3 weeks, the number of Mycobacterium tuberculosis in the lung was measured, and the defensive efficacy against Mycobacterium tuberculosis was confirmed. Information for PBS or DNA vaccine dosing groups are as follows: group 1, pbs (dosage form control); group 2, GX-70 μg; group 3, GX-171A 4 μg+BD-121A4 μg; group 4, GX-171B 4 μg+BD-121A4 μg; and group 5, bcg (positive control group).

The results confirmed that the number of Mycobacterium tuberculosis was lower in the administration group of all vaccine candidate substances than in the dosage form control group, and the degree of inflammation was also suppressed in the lung histopathological results (FIG. 3). However, the differences in defensive efficacy between individual vaccine candidate substances have not been confirmed.

To this end, the present inventors conducted further experiments to evaluate the efficacy of preventing tuberculosis by administering additionally prepared GX-170A and GX-170B in combination. In short, using OrbiJector(SL VAXiGEN, korea) in vivo electroporation apparatus PBS (dosage form control) or DNA vaccine was administered on thigh muscle of C57BL/6 mice 3 times every 3 weeks. In the BCG group of the positive control group, the C57BL/6 mice were administered once at week 3. After 3 weeks of final administration, mycobacterium tuberculosis was released into the air, and after 3 weeks, the number of Mycobacterium tuberculosis in the lung was measured, confirming the preventive effect on Mycobacterium tuberculosis. Information for PBS or DNA vaccine dosing groups are as follows: group 1, pbs (dosage form control); group 2, GX-70 μg; group 3, GX-70 μg+BD-121A8 μg; group 4, GX-171A8 μg+BD-121A8 μg; group 5, GX-170A 8. Mu.g+GX-170B 8. Mu.g+BD-121A 4. Mu.gg; and group 6, bcg (positive control group).

The results confirmed that, among the administration groups of all vaccine candidate substances, there was a significant effect of preventing tuberculosis as compared with the dosage form control group (fig. 4). In the group to which GX-70 was administered, it was confirmed that the effect of preventing tuberculosis was obtained regardless of the addition or non-addition of an immunoadjuvant, as compared with the dosage form control group, and in the group to which GX-171A and BD-121A were administered in combination, it was confirmed that the effect was similar to that of the group to which GX-70 was administered (FIG. 4). In particular, it was confirmed that in the group to which GX-170A, GX-170B and BD-121A were administered in combination, the effect of preventing tuberculosis was more remarkable than in the group to which GX-70 was administered, and the same effect was observed in the pathological results of lung tissue (FIG. 4).

EXAMPLE 5 analysis of treatment efficacy of tuberculosis

The present inventors have conducted experiments to confirm whether or not a composition containing the fusion protein and an immunoadjuvant has an effect of treating tuberculosis. Briefly, after releasing mycobacterium tuberculosis into the air, after 4 weeks, agents INH (isoniazid) and PZA (pyrazinamid e) for tuberculosis were administered to C57BL/6 mice, and treated with drinking water for 14 weeks. During this treatment period, BCG group (positive control group) was administered 1 time every 8 weeks, and ori was used(SL VAXiGEN, korea) in vivo electroporation apparatus PBS (dosage form control) or DNA vaccine was administered on thigh muscle of C57BL/6 mice 3 times every 5, 8, 11 and 13 weeks. After confirming that no mycobacterium tuberculosis was detected at the end time point of the tuberculosis drug treatment, the mycobacterium tuberculosis was released into the air a second time at week 20, after which the number of reinfected mycobacterium tuberculosis was observed in the lungs. The information of the group to which the DNA vaccine was administered was as follows: group 1, naiv without tuberculosis agent administered; group 2, pbs (dosage form control); group 3, GX-70 μg; group 4, GX-170A8 μg; group 5, GX-170A8 μg+GX-170B8 μg; group 6, GX-70 μg+BD-121A8 μg; group 7, GX-170A8 μg+BD-121A8 μg; group 8, GX-170A8 μg+GX-170B8 μg+BD-121A8 μg.

The results confirm that in all groups, tuberculosis re-infection has a lower incidence compared to untreated groups after infection with tuberculosis (fig. 5). In the group to which GX-170A and GX-170B were administered in combination, the number of Mycobacterium tuberculosis tended to decrease, and in particular, in the group to which GX-170A, GX-170B and BD-121A were administered in combination, it was confirmed that the number of Mycobacterium tuberculosis was significantly lower than that in the dosage form control group (FIG. 5).

From the above results, the present inventors confirmed that there is an efficacy of treating tuberculosis when GX-170A and GX-170B are administered in combination, and confirmed that the efficacy of treating tuberculosis is further improved when an additional immunoadjuvant is administered in combination.

<110> Gernacini Co., ltd

Disease management hall

<120> pharmaceutical composition for preventing or treating tuberculosis

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Asn Ile Asn Thr Lys Leu Gly Tyr Asn Asn Ala Val Gly Ala Gly Thr

35 40 45

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

50 55 60

Ile Ala Gly Ala Thr Asp Ile Asn Ala Phe Ser Val Gly Ser Gly Gln

65 70 75 80

Thr Tyr Gly Val Asp Val Val Gly Tyr Asp Arg Thr Gln Asn Val Ala

85 90 95

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

100 105 110

Gly Gly Val Ala Val Gly Glu Pro Val Val Ala Met Gly Asn Ser Gly

115 120 125

Gly Gln Gly Gly Thr Pro Arg Ala Val Pro Gly Arg Val Val Ala Leu

130 135 140

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

145 150 155 160

Leu Asn Gly Leu Ile Gln Phe Asp Ala Ala Ile Gln Pro Gly Asp Ala

165 170 175

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

180 185 190

Ala Ala Ser Asp Asn Phe Gln Leu Ser Gln Gly Gly Gln Gly Phe Ala

195 200 205

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

210 215 220

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

225 230 235 240

Gly Val Val Asp Asn Asn Gly Asn Gly Ala Arg Val Gln Arg Val Val

245 250 255

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

260 265 270

Thr Ala Val Asp Gly Ala Pro Ile Asn Ser Ala Thr Ala Met Ala Asp

275 280 285

Ala Leu Asn Gly His His Pro Gly Asp Val Ile Ser Val Thr Trp Gln

290 295 300

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

305 310 315 320

Pro Pro Ala

<210> 4

<211> 347

<212> PRT

<213> artificial sequence

<220>

<223> amino acid sequence of PstS3

<400> 4

Cys Gly Asn Asp Asp Asn Val Thr Gly Gly Gly Ala Thr Thr Gly Gln

1 5 10 15

Ala Ser Ala Lys Val Asp Cys Gly Gly Lys Lys Thr Leu Lys Ala Ser

20 25 30

Gly Ser Thr Ala Gln Ala Asn Ala Met Thr Arg Phe Val Asn Val Phe

35 40 45

Glu Gln Ala Cys Pro Gly Gln Thr Leu Asn Tyr Thr Ala Asn Gly Ser

50 55 60

Gly Ala Gly Ile Ser Glu Phe Asn Gly Asn Gln Thr Asp Phe Gly Gly

65 70 75 80

Ser Asp Val Pro Leu Ser Lys Asp Glu Pro Ser Gly Ala Ala Arg Cys

85 90 95

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

100 105 110

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

115 120 125

Leu Ala Lys Ile Phe Asn Gly Ser Ile Thr Gln Trp Asn Asn Pro Ala

130 135 140

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

145 150 155 160

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

165 170 175

Tyr Leu Gln Ala Ala Ser Asn Gly Ala Trp Gly Lys Gly Ala Gly Lys

180 185 190

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

195 200 205

Ser Ala Ala Ala Lys Asn Thr Pro Gly Ser Ile Thr Tyr Asn Glu Trp

210 215 220

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

225 230 235 240

Ala Gly Gly Asp Pro Val Ala Ile Thr Ile Asp Ser Val Gly Gln Thr

245 250 255

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

260 265 270

Thr Asp Ser Phe Tyr Arg Pro Lys Arg Pro Gly Ser Tyr Pro Ile Val

275 280 285

Leu Ala Thr Tyr Glu Ile Val Cys Ser Lys Tyr Pro Asp Ser Gln Val

290 295 300

Gly Thr Ala Val Lys Ala Phe Leu Gln Ser Thr Ile Gly Ala Gly Gln

305 310 315 320

Ser Gly Leu Gly Asp Asn Gly Tyr Ile Pro Ile Pro Asp Glu Phe Lys

325 330 335

Ser Arg Leu Ser Thr Ala Val Asn Ala Ile Ala

340 345

<210> 5

<211> 621

<212> PRT

<213> artificial sequence

<220>

<223> amino acid sequence of PPE39

<400> 5

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

1 5 10 15

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

20 25 30

Gly Leu Ala Ala Glu Leu Gly Leu Ala Ala Glu Ser Phe Gly Leu Val

35 40 45

Thr Ser Gly Leu Ala Gly Gly Ser Gly Gln Ala Trp Gln Gly Ala Ala

50 55 60

Ala Ala Ala Met Val Val Ala Ala Ala Pro Tyr Ala Gly Trp Leu Ala

65 70 75 80

Ala Ala Ala Ala Arg Ala Gly Gly Ala Ala Val Gln Ala Lys Ala Val

85 90 95

Ala Gly Ala Phe Glu Ala Ala Arg Ala Ala Met Val Asp Pro Val Val

100 105 110

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

115 120 125

Phe Gly Gln Asn Ala Pro Ala Ile Ala Ala Ala Glu Ala Thr Tyr Glu

130 135 140

Gln Met Trp Ala Ala Asp Val Ala Ala Met Val Gly Tyr His Gly Gly

145 150 155 160

Ala Ser Ala Ala Ala Ala Ala Leu Ala Pro Trp Gln Gln Ala Val Pro

165 170 175

Gly Leu Ser Gly Leu Leu Asp Ser Ala Gln Ser Ser Ala Gln Ala Val

180 185 190

Thr Ala Gln Ala Val Gly Ser Thr Val Pro Gly Pro Leu Gln Gly Ile

195 200 205

Asn Phe Gly Phe Gly Asn Ile Gly Ser Leu Asn Leu Gly Ser Gly Asn

210 215 220

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

225 230 235 240

Gly Gly Gly Asn Ile Gly Ser Phe Asn Leu Gly Ser Gly Asn Gln Gly

245 250 255

Asp Ile Asn Leu Gly Ile Gly Asn Val Gly Asn Leu Asn Leu Gly Ser

260 265 270

Gly Asn Phe Gly Ser Gln Asn Leu Gly Ser Gly Asn Ile Gly Ser Thr

275 280 285

Asn Val Gly Ser Gly Asn Ile Gly Ser Thr Asn Val Gly Ser Gly Asn

290 295 300

Ile Gly Asp Thr Asn Phe Gly Asn Gly Asn Asn Gly Asn Phe Asn Phe

305 310 315 320

Gly Ser Gly Asn Thr Gly Ser Asn Asn Ile Gly Phe Gly Asn Thr Gly

325 330 335

Ser Gly Asn Phe Gly Phe Gly Asn Thr Gly Asn Asn Asn Ile Gly Ile

340 345 350

Gly Leu Thr Gly Asp Gly Gln Ile Gly Ile Gly Gly Leu Asn Ser Gly

355 360 365

Ser Gly Asn Ile Gly Phe Gly Asn Ser Gly Thr Gly Asn Val Gly Leu

370 375 380

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

385 390 395 400

Asn Thr Gly Phe Gly Asn Ala Gly Asn Val Asn Thr Gly Phe Trp Asn

405 410 415

Gly Gly Ser Thr Asn Thr Gly Leu Ala Asn Ala Gly Ala Gly Asn Thr

420 425 430

Gly Phe Phe Asp Ala Gly Asn Tyr Asn Phe Gly Ser Leu Asn Ala Gly

435 440 445

Asn Ile Asn Ser Ser Phe Gly Asn Ser Gly Asp Gly Asn Ser Gly Phe

450 455 460

Leu Asn Ala Gly Asp Val Asn Ser Gly Val Gly Asn Ala Gly Asp Val

465 470 475 480

Asn Thr Gly Leu Gly Asn Ser Gly Asn Ile Asn Thr Gly Gly Phe Asn

485 490 495

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

500 505 510

Pro Asn Ser Gly Phe Phe Asn Ala Gly Thr Gly Asn Ser Gly Phe Gly

515 520 525

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

530 535 540

Phe Gly Asn Ser Gly Tyr Val Asn Thr Ser Thr Thr Ser Met Phe Gly

545 550 555 560

Gly Asn Ser Gly Val Leu Asn Thr Gly Tyr Gly Asn Ser Gly Phe Tyr

565 570 575

Asn Ala Ala Val Asn Asn Thr Gly Ile Phe Val Thr Gly Val Met Ser

580 585 590

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

595 600 605

Gly Asn Gly Leu Ser Gly Phe Phe Lys Asn Leu Phe Gly

610 615 620

<210> 6

<211> 740

<212> PRT

<213> artificial sequence

<220>

<223> amino acid sequence of GlcB

<400> 6

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

1 5 10 15

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

20 25 30

Ser Phe Trp Ala Gly Val Asp Lys Val Val Ala Asp Leu Thr Pro Gln

35 40 45

Asn Gln Ala Leu Leu Asn Ala Arg Asp Glu Leu Gln Ala Gln Ile Asp

50 55 60

Lys Trp His Arg Arg Arg Val Ile Glu Pro Ile Asp Met Asp Ala Tyr

65 70 75 80

Arg Gln Phe Leu Thr Glu Ile Gly Tyr Leu Leu Pro Glu Pro Asp Asp

85 90 95

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

100 105 110

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

115 120 125

Ala Ala Asn Ala Arg Trp Gly Ser Leu Tyr Asp Ala Leu Tyr Gly Thr

130 135 140

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

145 150 155 160

Lys Val Arg Gly Asp Lys Val Ile Ala Tyr Ala Arg Lys Phe Leu Asp

165 170 175

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

180 185 190

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

195 200 205

Gly Leu Ala Asn Pro Gly Gln Phe Ala Gly Tyr Thr Gly Ala Ala Glu

210 215 220

Ser Pro Thr Ser Val Leu Leu Ile Asn His Gly Leu His Ile Glu Ile

225 230 235 240

Leu Ile Asp Pro Glu Ser Gln Val Gly Thr Thr Asp Arg Ala Gly Val

245 250 255

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

260 265 270

Asp Ser Val Ala Ala Val Asp Ala Ala Asp Lys Val Leu Gly Tyr Arg

275 280 285

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

290 295 300

Asp Gly Thr Ala Phe Leu Arg Val Leu Asn Arg Asp Arg Asn Tyr Thr

305 310 315 320

Ala Pro Gly Gly Gly Gln Phe Thr Leu Pro Gly Arg Ser Leu Met Phe

325 330 335

Val Arg Asn Val Gly His Leu Met Thr Asn Asp Ala Ile Val Asp Thr

340 345 350

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

355 360 365

Leu Ile Ala Ile His Gly Leu Lys Ala Ser Asp Val Asn Gly Pro Leu

370 375 380

Ile Asn Ser Arg Thr Gly Ser Ile Tyr Ile Val Lys Pro Lys Met His

385 390 395 400

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

405 410 415

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

420 425 430

Glu Glu Arg Arg Thr Thr Val Asn Leu Lys Ala Cys Ile Lys Ala Ala

435 440 445

Ala Asp Arg Val Val Phe Ile Asn Thr Gly Phe Leu Asp Arg Thr Gly

450 455 460

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

465 470 475 480

Thr Met Lys Ser Gln Pro Trp Ile Leu Ala Tyr Glu Asp His Asn Val

485 490 495

Asp Ala Gly Leu Ala Ala Gly Phe Ser Gly Arg Ala Gln Val Gly Lys

500 505 510

Gly Met Trp Thr Met Thr Glu Leu Met Ala Asp Met Val Glu Thr Lys

515 520 525

Ile Ala Gln Pro Arg Ala Gly Ala Ser Thr Ala Trp Val Pro Ser Pro

530 535 540

Thr Ala Ala Thr Leu His Ala Leu His Tyr His Gln Val Asp Val Ala

545 550 555 560

Ala Val Gln Gln Gly Leu Ala Gly Lys Arg Arg Ala Thr Ile Glu Gln

565 570 575

Leu Leu Thr Ile Pro Leu Ala Lys Glu Leu Ala Trp Ala Pro Asp Glu

580 585 590

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

595 600 605

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

610 615 620

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

625 630 635 640

Gln Leu Leu Ala Asn Trp Leu Arg His Gly Val Ile Thr Ser Ala Asp

645 650 655

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

660 665 670

Ala Gly Asp Val Ala Tyr Arg Pro Met Ala Pro Asn Phe Asp Asp Ser

675 680 685

Ile Ala Phe Leu Ala Ala Gln Glu Leu Ile Leu Ser Gly Ala Gln Gln

690 695 700

Pro Asn Gly Tyr Thr Glu Pro Ile Leu His Arg Arg Arg Arg Glu Phe

705 710 715 720

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

725 730 735

Asp Ala Ala Arg

740

<210> 7

<211> 433

<212> PRT

<213> artificial sequence

<220>

<223> amino acid sequence of RipA

<400> 7

Asp Pro Gln Thr Asp Thr Ile Ala Ala Leu Ile Ala Asp Val Ala Lys

1 5 10 15

Ala Asn Gln Arg Leu Gln Asp Leu Ser Asp Glu Val Gln Ala Glu Gln

20 25 30

Glu Ser Val Asn Lys Ala Met Val Asp Val Glu Thr Ala Arg Asp Asn

35 40 45

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

50 55 60

Asp Ala Asn Ala Ala Ile Ala Ala Ala Gln His Arg Phe Asp Thr Phe

65 70 75 80

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

85 90 95

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

100 105 110

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

115 120 125

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

130 135 140

Asp Lys Ala Ala Ala Asp Ala Lys Ala Ser Gln Asp Ala Ala Val Ala

145 150 155 160

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

165 170 175

Gln Arg Leu Ala Ala Glu Arg Asp Ala Ala Gln Ala Arg Leu Gln Ala

180 185 190

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

195 200 205

Phe Arg Met Trp Asp Pro Gly Ser Gly Pro Ala Gly Gly Arg Ala Trp

210 215 220

Asp Gly Leu Trp Asp Pro Thr Leu Pro Met Ile Pro Ser Ala Asn Ile

225 230 235 240

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

245 250 255

Thr Ser Ala Gln Val Thr Ala Asn Met Gly Arg Lys Phe Leu Glu Gln

260 265 270

Leu Gly Ile Leu Gln Pro Thr Asp Thr Gly Ile Thr Asn Ala Pro Ala

275 280 285

Gly Ser Ala Gln Gly Arg Ile Pro Arg Val Tyr Gly Arg Gln Ala Ser

290 295 300

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

305 310 315 320

Trp Gly Gly Gly Asn Ala Ala Gly Pro Ser Lys Gly Ile Asp Ser Gly

325 330 335

Ala Gly Thr Val Gly Phe Asp Cys Ser Gly Leu Val Leu Tyr Ser Phe

340 345 350

Ala Gly Val Gly Ile Lys Leu Pro His Tyr Ser Gly Ser Gln Tyr Asn

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

Pro Val Arg Thr Ala Gly Met Thr Pro Tyr Val Val Arg Tyr Ile Glu

420 425 430

Tyr

<210> 8

<211> 10

<212> PRT

<213> artificial sequence

<220>

<223> amino acid sequence of GS linker

<400> 8

Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser

1 5 10

<210> 9

<211> 23

<212> PRT

<213> artificial sequence

<220>

<223> amino acid sequence of tPA

<400> 9

Met Asp Ala Met Lys Arg Gly Leu Cys Cys Val Leu Leu Leu Cys Gly

1 5 10 15

Ala Val Phe Val Ser Pro Ser

20

<210> 10

<211> 156

<212> PRT

<213> artificial sequence

<220>

<223> amino acid sequence of Flt3L

<400> 10

Thr Gln Asp Cys Ser Phe Gln His Ser Pro Ile Ser Ser Asp Phe Ala

1 5 10 15

Val Lys Ile Arg Glu Leu Ser Asp Tyr Leu Leu Gln Asp Tyr Pro Val

20 25 30

Thr Val Ala Ser Asn Leu Gln Asp Glu Glu Leu Cys Gly Gly Leu Trp

35 40 45

Arg Leu Val Leu Ala Gln Arg Trp Met Glu Arg Leu Lys Thr Val Ala

50 55 60

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

65 70 75 80

Phe Val Thr Lys Cys Ala Phe Gln Pro Pro Pro Ser Cys Leu Arg Phe

85 90 95

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

100 105 110

Val Ala Leu Lys Pro Trp Ile Thr Arg Gln Asn Phe Ser Arg Cys Leu

115 120 125

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

130 135 140

Pro Arg Pro Leu Glu Ala Thr Ala Pro Thr Ala Pro

145 150 155

<210> 11

<211> 1281

<212> PRT

<213> artificial sequence

<220>

<223> amino acid sequence of GX-170A

<400> 11

Met Asp Ala Met Lys Arg Gly Leu Cys Cys Val Leu Leu Leu Cys Gly

1 5 10 15

Ala Val Phe Val Ser Pro Ser His Ala Thr Gln Asp Cys Ser Phe Gln

20 25 30

His Ser Pro Ile Ser Ser Asp Phe Ala Val Lys Ile Arg Glu Leu Ser

35 40 45

Asp Tyr Leu Leu Gln Asp Tyr Pro Val Thr Val Ala Ser Asn Leu Gln

50 55 60

Asp Glu Glu Leu Cys Gly Gly Leu Trp Arg Leu Val Leu Ala Gln Arg

65 70 75 80

Trp Met Glu Arg Leu Lys Thr Val Ala Gly Ser Lys Met Gln Gly Leu

85 90 95

Leu Glu Arg Val Asn Thr Glu Ile His Phe Val Thr Lys Cys Ala Phe

100 105 110

Gln Pro Pro Pro Ser Cys Leu Arg Phe Val Gln Thr Asn Ile Ser Arg

115 120 125

Leu Leu Gln Glu Thr Ser Glu Gln Leu Val Ala Leu Lys Pro Trp Ile

130 135 140

Thr Arg Gln Asn Phe Ser Arg Cys Leu Glu Leu Gln Cys Gln Pro Asp

145 150 155 160

Ser Ser Thr Leu Pro Pro Pro Trp Ser Pro Arg Pro Leu Glu Ala Thr

165 170 175

Ala Pro Thr Ala Pro Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Phe

180 185 190

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

195 200 205

Met Gly Arg Asp Ile Lys Val Gln Phe Gln Ser Gly Gly Ala Asn Ser

210 215 220

Pro Ala Leu Tyr Leu Leu Asp Gly Leu Arg Ala Gln Asp Asp Phe Ser

225 230 235 240

Gly Trp Asp Ile Asn Thr Pro Ala Phe Glu Trp Tyr Asp Gln Ser Gly

245 250 255

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

260 265 270

Trp Tyr Gln Pro Ala Cys Gly Lys Ala Gly Cys Gln Thr Tyr Lys Trp

275 280 285

Glu Thr Phe Leu Thr Ser Glu Leu Pro Gly Trp Leu Gln Ala Asn Arg

290 295 300

His Val Lys Pro Thr Gly Ser Ala Val Val Gly Leu Ser Met Ala Ala

305 310 315 320

Ser Ser Ala Leu Thr Leu Ala Ile Tyr His Pro Gln Gln Phe Val Tyr

325 330 335

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

340 345 350

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

355 360 365

Asp Met Trp Gly Pro Lys Glu Asp Pro Ala Trp Gln Arg Asn Asp Pro

370 375 380

Leu Leu Asn Val Gly Lys Leu Ile Ala Asn Asn Thr Arg Val Trp Val

385 390 395 400

Tyr Cys Gly Asn Gly Lys Pro Ser Asp Leu Gly Gly Asn Asn Leu Pro

405 410 415

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

420 425 430

Asp Ala Tyr Asn Ala Gly Gly Gly His Asn Gly Val Phe Asp Phe Pro

435 440 445

Asp Ser Gly Thr His Ser Trp Glu Tyr Trp Gly Ala Gln Leu Asn Ala

450 455 460

Met Lys Pro Asp Leu Gln Arg Ala Leu Gly Ala Thr Pro Asn Thr Gly

465 470 475 480

Pro Ala Pro Gln Gly Ala Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser

485 490 495

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

500 505 510

Met Ala Gly Tyr Ala Gly Thr Leu Gln Ser Leu Gly Ala Glu Ile Ala

515 520 525

Val Glu Gln Ala Ala Leu Gln Ser Ala Trp Gln Gly Asp Thr Gly Ile

530 535 540

Thr Tyr Gln Ala Trp Gln Ala Gln Trp Asn Gln Ala Met Glu Asp Leu

545 550 555 560

Val Arg Ala Tyr His Ala Met Ser Ser Thr His Glu Ala Asn Thr Met

565 570 575

Ala Met Met Ala Arg Asp Thr Ala Glu Ala Ala Lys Trp Gly Gly Gly

580 585 590

Ser Gly Ser Gly Ser Gly Ser Gly Ser Ala Pro Pro Ala Leu Ser Gln

595 600 605

Asp Arg Phe Ala Asp Phe Pro Ala Leu Pro Leu Asp Pro Ser Ala Met

610 615 620

Val Ala Gln Val Gly Pro Gln Val Val Asn Ile Asn Thr Lys Leu Gly

625 630 635 640

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

645 650 655

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

660 665 670

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

675 680 685

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

690 695 700

Gly Gly Leu Pro Ser Ala Ala Ile Gly Gly Gly Val Ala Val Gly Glu

705 710 715 720

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

725 730 735

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

740 745 750

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

755 760 765

Asp Ala Ala Ile Gln Pro Gly Asp Ala Gly Gly Pro Val Val Asn Gly

770 775 780

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

785 790 795 800

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

805 810 815

Ala Ile Ala Gly Gln Ile Arg Ser Gly Gly Gly Ser Pro Thr Val His

820 825 830

Ile Gly Pro Thr Ala Phe Leu Gly Leu Gly Val Val Asp Asn Asn Gly

835 840 845

Asn Gly Ala Arg Val Gln Arg Val Val Gly Ser Ala Pro Ala Ala Ser

850 855 860

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

865 870 875 880

Ile Asn Ser Ala Thr Ala Met Ala Asp Ala Leu Asn Gly His His Pro

885 890 895

Gly Asp Val Ile Ser Val Thr Trp Gln Thr Lys Ser Gly Gly Thr Arg

900 905 910

Thr Gly Asn Val Thr Leu Ala Glu Gly Pro Pro Ala Gly Ser Gly Ser

915 920 925

Gly Ser Gly Ser Gly Ser Cys Gly Asn Asp Asp Asn Val Thr Gly Gly

930 935 940

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

945 950 955 960

Lys Thr Leu Lys Ala Ser Gly Ser Thr Ala Gln Ala Asn Ala Met Thr

965 970 975

Arg Phe Val Asn Val Phe Glu Gln Ala Cys Pro Gly Gln Thr Leu Asn

980 985 990

Tyr Thr Ala Asn Gly Ser Gly Ala Gly Ile Ser Glu Phe Asn Gly Asn

995 1000 1005

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

1010 1015 1020

Ser Gly Ala Ala Arg Cys Gly Ser Pro Ala Trp Asn Leu Pro Val Val

1025 1030 1035 1040

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

1045 1050 1055

Asn Leu Asp Gly Pro Thr Leu Ala Lys Ile Phe Asn Gly Ser Ile Thr

1060 1065 1070

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

1075 1080 1085

Pro Gly Glu Arg Ile His Val Val Phe Arg Ser Asp Glu Ser Gly Thr

1090 1095 1100

Thr Asp Asn Phe Gln Arg Tyr Leu Gln Ala Ala Ser Asn Gly Ala Trp

1105 1110 1115 1120

Gly Lys Gly Ala Gly Lys Ser Phe Gln Gly Gly Val Gly Glu Gly Ala

1125 1130 1135

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

1140 1145 1150

Ile Thr Tyr Asn Glu Trp Ser Phe Ala Gln Ala His Asp Leu Thr Met

1155 1160 1165

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

1170 1175 1180

Asp Ser Val Gly Gln Thr Ile Ala Gly Ala Thr Ile Ser Gly Val Gly

1185 1190 1195 1200

Asn Asp Leu Val Leu Asp Thr Asp Ser Phe Tyr Arg Pro Lys Arg Pro

1205 1210 1215

Gly Ser Tyr Pro Ile Val Leu Ala Thr Tyr Glu Ile Val Cys Ser Lys

1220 1225 1230

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

1235 1240 1245

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

1250 1255 1260

Ile Pro Asp Glu Phe Lys Ser Arg Leu Ser Thr Ala Val Asn Ala Ile

1265 1270 1275 1280

Ala

<210> 12

<211> 2005

<212> PRT

<213> artificial sequence

<220>

<223> amino acid sequence of GX-170B

<400> 12

Met Asp Ala Met Lys Arg Gly Leu Cys Cys Val Leu Leu Leu Cys Gly

1 5 10 15

Ala Val Phe Val Ser Pro Ser His Ala Thr Gln Asp Cys Ser Phe Gln

20 25 30

His Ser Pro Ile Ser Ser Asp Phe Ala Val Lys Ile Arg Glu Leu Ser

35 40 45

Asp Tyr Leu Leu Gln Asp Tyr Pro Val Thr Val Ala Ser Asn Leu Gln

50 55 60

Asp Glu Glu Leu Cys Gly Gly Leu Trp Arg Leu Val Leu Ala Gln Arg

65 70 75 80

Trp Met Glu Arg Leu Lys Thr Val Ala Gly Ser Lys Met Gln Gly Leu

85 90 95

Leu Glu Arg Val Asn Thr Glu Ile His Phe Val Thr Lys Cys Ala Phe

100 105 110

Gln Pro Pro Pro Ser Cys Leu Arg Phe Val Gln Thr Asn Ile Ser Arg

115 120 125

Leu Leu Gln Glu Thr Ser Glu Gln Leu Val Ala Leu Lys Pro Trp Ile

130 135 140

Thr Arg Gln Asn Phe Ser Arg Cys Leu Glu Leu Gln Cys Gln Pro Asp

145 150 155 160

Ser Ser Thr Leu Pro Pro Pro Trp Ser Pro Arg Pro Leu Glu Ala Thr

165 170 175

Ala Pro Thr Ala Pro Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Val

180 185 190

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

195 200 205

Gly Ala Gly Ser Gly Pro Met Leu Ala Ala Ala Ala Ala Trp Asp Gly

210 215 220

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

225 230 235 240

Ser Gly Leu Ala Gly Gly Ser Gly Gln Ala Trp Gln Gly Ala Ala Ala

245 250 255

Ala Ala Met Val Val Ala Ala Ala Pro Tyr Ala Gly Trp Leu Ala Ala

260 265 270

Ala Ala Ala Arg Ala Gly Gly Ala Ala Val Gln Ala Lys Ala Val Ala

275 280 285

Gly Ala Phe Glu Ala Ala Arg Ala Ala Met Val Asp Pro Val Val Val

290 295 300

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

305 310 315 320

Gly Gln Asn Ala Pro Ala Ile Ala Ala Ala Glu Ala Thr Tyr Glu Gln

325 330 335

Met Trp Ala Ala Asp Val Ala Ala Met Val Gly Tyr His Gly Gly Ala

340 345 350

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

355 360 365

Leu Ser Gly Leu Leu Asp Ser Ala Gln Ser Ser Ala Gln Ala Val Thr

370 375 380

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

385 390 395 400

Phe Gly Phe Gly Asn Ile Gly Ser Leu Asn Leu Gly Ser Gly Asn Thr

405 410 415

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

420 425 430

Gly Gly Asn Ile Gly Ser Phe Asn Leu Gly Ser Gly Asn Gln Gly Asp

435 440 445

Ile Asn Leu Gly Ile Gly Asn Val Gly Asn Leu Asn Leu Gly Ser Gly

450 455 460

Asn Phe Gly Ser Gln Asn Leu Gly Ser Gly Asn Ile Gly Ser Thr Asn

465 470 475 480

Val Gly Ser Gly Asn Ile Gly Ser Thr Asn Val Gly Ser Gly Asn Ile

485 490 495

Gly Asp Thr Asn Phe Gly Asn Gly Asn Asn Gly Asn Phe Asn Phe Gly

500 505 510

Ser Gly Asn Thr Gly Ser Asn Asn Ile Gly Phe Gly Asn Thr Gly Ser

515 520 525

Gly Asn Phe Gly Phe Gly Asn Thr Gly Asn Asn Asn Ile Gly Ile Gly

530 535 540

Leu Thr Gly Asp Gly Gln Ile Gly Ile Gly Gly Leu Asn Ser Gly Ser

545 550 555 560

Gly Asn Ile Gly Phe Gly Asn Ser Gly Thr Gly Asn Val Gly Leu Phe

565 570 575

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

580 585 590

Thr Gly Phe Gly Asn Ala Gly Asn Val Asn Thr Gly Phe Trp Asn Gly

595 600 605

Gly Ser Thr Asn Thr Gly Leu Ala Asn Ala Gly Ala Gly Asn Thr Gly

610 615 620

Phe Phe Asp Ala Gly Asn Tyr Asn Phe Gly Ser Leu Asn Ala Gly Asn

625 630 635 640

Ile Asn Ser Ser Phe Gly Asn Ser Gly Asp Gly Asn Ser Gly Phe Leu

645 650 655

Asn Ala Gly Asp Val Asn Ser Gly Val Gly Asn Ala Gly Asp Val Asn

660 665 670

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

675 680 685

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

690 695 700

Asn Ser Gly Phe Phe Asn Ala Gly Thr Gly Asn Ser Gly Phe Gly His

705 710 715 720

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

725 730 735

Gly Asn Ser Gly Tyr Val Asn Thr Ser Thr Thr Ser Met Phe Gly Gly

740 745 750

Asn Ser Gly Val Leu Asn Thr Gly Tyr Gly Asn Ser Gly Phe Tyr Asn

755 760 765

Ala Ala Val Asn Asn Thr Gly Ile Phe Val Thr Gly Val Met Ser Ser

770 775 780

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

785 790 795 800

Asn Gly Leu Ser Gly Phe Phe Lys Asn Leu Phe Gly Gly Ser Gly Ser

805 810 815

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

820 825 830

Ile Ala Arg Val Leu Tyr Asp Phe Val Asn Asn Glu Ala Leu Pro Gly

835 840 845

Thr Asp Ile Asp Pro Asp Ser Phe Trp Ala Gly Val Asp Lys Val Val

850 855 860

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

865 870 875 880

Leu Gln Ala Gln Ile Asp Lys Trp His Arg Arg Arg Val Ile Glu Pro

885 890 895

Ile Asp Met Asp Ala Tyr Arg Gln Phe Leu Thr Glu Ile Gly Tyr Leu

900 905 910

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

915 920 925

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

930 935 940

Ala Arg Phe Ala Leu Asn Ala Ala Asn Ala Arg Trp Gly Ser Leu Tyr

945 950 955 960

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

965 970 975

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

980 985 990

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

995 1000 1005

Gly Asp Ala Thr Gly Phe Thr Val Gln Asp Gly Gln Leu Val Val Ala

1010 1015 1020

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

1025 1030 1035 1040

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

1045 1050 1055

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

1060 1065 1070

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

1075 1080 1085

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

1090 1095 1100

Lys Val Leu Gly Tyr Arg Asn Trp Leu Gly Leu Asn Lys Gly Asp Leu

1105 1110 1115 1120

Ala Ala Ala Val Asp Lys Asp Gly Thr Ala Phe Leu Arg Val Leu Asn

1125 1130 1135

Arg Asp Arg Asn Tyr Thr Ala Pro Gly Gly Gly Gln Phe Thr Leu Pro

1140 1145 1150

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

1155 1160 1165

Asp Ala Ile Val Asp Thr Asp Gly Ser Glu Val Phe Glu Gly Ile Met

1170 1175 1180

Asp Ala Leu Phe Thr Gly Leu Ile Ala Ile His Gly Leu Lys Ala Ser

1185 1190 1195 1200

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

1205 1210 1215

Val Lys Pro Lys Met His Gly Pro Ala Glu Val Ala Phe Thr Cys Glu

1220 1225 1230

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

1235 1240 1245

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

1250 1255 1260

Ala Cys Ile Lys Ala Ala Ala Asp Arg Val Val Phe Ile Asn Thr Gly

1265 1270 1275 1280

Phe Leu Asp Arg Thr Gly Asp Glu Ile His Thr Ser Met Glu Ala Gly

1285 1290 1295

Pro Met Val Arg Lys Gly Thr Met Lys Ser Gln Pro Trp Ile Leu Ala

1300 1305 1310

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

1315 1320 1325

Arg Ala Gln Val Gly Lys Gly Met Trp Thr Met Thr Glu Leu Met Ala

1330 1335 1340

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

1345 1350 1355 1360

Ala Trp Val Pro Ser Pro Thr Ala Ala Thr Leu His Ala Leu His Tyr

1365 1370 1375

His Gln Val Asp Val Ala Ala Val Gln Gln Gly Leu Ala Gly Lys Arg

1380 1385 1390

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

1395 1400 1405

Ala Trp Ala Pro Asp Glu Ile Arg Glu Glu Val Asp Asn Asn Cys Gln

1410 1415 1420

Ser Ile Leu Gly Tyr Val Val Arg Trp Val Asp Gln Gly Val Gly Cys

1425 1430 1435 1440

Ser Lys Val Pro Asp Ile His Asp Val Ala Leu Met Glu Asp Arg Ala

1445 1450 1455

Thr Leu Arg Ile Ser Ser Gln Leu Leu Ala Asn Trp Leu Arg His Gly

1460 1465 1470

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

1475 1480 1485

Leu Val Asp Arg Gln Asn Ala Gly Asp Val Ala Tyr Arg Pro Met Ala

1490 1495 1500

Pro Asn Phe Asp Asp Ser Ile Ala Phe Leu Ala Ala Gln Glu Leu Ile

1505 1510 1515 1520

Leu Ser Gly Ala Gln Gln Pro Asn Gly Tyr Thr Glu Pro Ile Leu His

1525 1530 1535

Arg Arg Arg Arg Glu Phe Lys Ala Arg Ala Ala Glu Lys Pro Ala Pro

1540 1545 1550

Ser Asp Arg Ala Gly Asp Asp Ala Ala Arg Gly Ser Gly Ser Gly Ser

1555 1560 1565

Gly Ser Gly Ser Asp Pro Gln Thr Asp Thr Ile Ala Ala Leu Ile Ala

1570 1575 1580

Asp Val Ala Lys Ala Asn Gln Arg Leu Gln Asp Leu Ser Asp Glu Val

1585 1590 1595 1600

Gln Ala Glu Gln Glu Ser Val Asn Lys Ala Met Val Asp Val Glu Thr

1605 1610 1615

Ala Arg Asp Asn Ala Ala Ala Ala Glu Asp Asp Leu Glu Val Ser Gln

1620 1625 1630

Arg Ala Val Lys Asp Ala Asn Ala Ala Ile Ala Ala Ala Gln His Arg

1635 1640 1645

Phe Asp Thr Phe Ala Ala Ala Thr Tyr Met Asn Gly Pro Ser Val Ser

1650 1655 1660

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

1665 1670 1675 1680

Ala Lys Thr Leu Ser Ala Ser Ser Gln Ala Val Met Ala Asn Leu Gln

1685 1690 1695

Arg Ala Arg Thr Glu Arg Val Asn Thr Glu Ser Ala Ala Arg Leu Ala

1700 1705 1710

Lys Gln Lys Ala Asp Lys Ala Ala Ala Asp Ala Lys Ala Ser Gln Asp

1715 1720 1725

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

1730 1735 1740

Arg Glu Glu Val Gln Arg Leu Ala Ala Glu Arg Asp Ala Ala Gln Ala

1745 1750 1755 1760

Arg Leu Gln Ala Ala Arg Leu Val Ala Trp Ser Ser Glu Gly Gly Gln

1765 1770 1775

Gly Ala Pro Pro Phe Arg Met Trp Asp Pro Gly Ser Gly Pro Ala Gly

1780 1785 1790

Gly Arg Ala Trp Asp Gly Leu Trp Asp Pro Thr Leu Pro Met Ile Pro

1795 1800 1805

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

1810 1815 1820

Gly Ile Ser Ala Thr Ser Ala Gln Val Thr Ala Asn Met Gly Arg Lys

1825 1830 1835 1840

Phe Leu Glu Gln Leu Gly Ile Leu Gln Pro Thr Asp Thr Gly Ile Thr

1845 1850 1855

Asn Ala Pro Ala Gly Ser Ala Gln Gly Arg Ile Pro Arg Val Tyr Gly

1860 1865 1870

Arg Gln Ala Ser Glu Tyr Val Ile Arg Arg Gly Met Ser Gln Ile Gly

1875 1880 1885

Val Pro Tyr Ser Trp Gly Gly Gly Asn Ala Ala Gly Pro Ser Lys Gly

1890 1895 1900

Ile Asp Ser Gly Ala Gly Thr Val Gly Phe Asp Cys Ser Gly Leu Val

1905 1910 1915 1920

Leu Tyr Ser Phe Ala Gly Val Gly Ile Lys Leu Pro His Tyr Ser Gly

1925 1930 1935

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

1940 1945 1950

Gly Asp Val Ile Phe Tyr Gly Pro Asn Gly Ser Gln His Val Thr Ile

1955 1960 1965

Tyr Leu Gly Asn Gly Gln Met Leu Glu Ala Pro Asp Val Gly Leu Lys

1970 1975 1980

Val Arg Val Ala Pro Val Arg Thr Ala Gly Met Thr Pro Tyr Val Val

1985 1990 1995 2000

Arg Tyr Ile Glu Tyr

2005

<210> 13

<211> 885

<212> DNA

<213> artificial sequence

<220>

<223> nucleic acid sequence of Ag85A

<400> 13

tttagcaggc caggcctgcc cgtggagtat ctgcaggtgc cctccccttc tatgggccgc 60

gacatcaagg tgcagttcca gtccggagga gccaactctc cagccctgta tctgctggac 120

ggcctgagag cccaggacga tttcagcggc tgggacatca atacacccgc ctttgagtgg 180

tacgaccaga gcggactgtc cgtggtcatg ccagtgggag gacagtcctc tttttactcc 240

gactggtatc agcctgcatg cggcaaggca ggatgtcaga cctacaagtg ggagaccttc 300

ctgacaagcg agctgcctgg ctggctgcag gccaacaggc acgtgaagcc aacaggctcc 360

gccgtggtgg gactgtctat ggcagccagc tccgccctga ccctggcaat ctaccaccct 420

cagcagttcg tgtacgcagg agcaatgagc ggactgctgg acccctccca ggcaatggga 480

ccaaccctga tcggactggc aatgggcgac gcaggaggat ataaggcaag cgacatgtgg 540

ggaccaaagg aggaccccgc atggcagagg aacgacccac tgctgaatgt gggcaagctg 600

atcgccaaca ataccagagt gtgggtgtac tgtggaaacg gcaagccatc cgatctggga 660

ggaaacaatc tgcctgccaa gttcctggag ggctttgtgc ggacatctaa tatcaagttc 720

caggacgcat ataacgcagg aggaggacac aatggcgtgt tcgactttcc cgattccggc 780

acccactctt gggagtactg gggagcacag ctgaacgcaa tgaagcccga cctgcagaga 840

gccctgggag caacccccaa tacaggacca gcaccacagg gagca 885

<210> 14

<211> 285

<212> DNA

<213> artificial sequence

<220>

<223> nucleic acid sequence of TB10.4

<400> 14

agccagatca tgtacaacta tcctgccatg ctgggacacg caggcgacat ggcaggatac 60

gcaggcaccc tgcagtctct gggagcagag atcgcagtgg agcaggccgc cctgcagagc 120

gcctggcagg gcgataccgg catcacatat caggcatggc aggcacagtg gaaccaggca 180

atggaggacc tggtgcgggc ctaccacgcc atgtcctcta cccacgaggc caatacaatg 240

gccatgatgg ccagagatac cgcagaggca gcaaagtggg gagga 285

<210> 15

<211> 969

<212> DNA

<213> artificial sequence

<220>

<223> nucleic acid sequence of Mtb32b

<400> 15

gccccccctg ccctgtccca ggaccggttc gccgattttc ctgccctgcc actggacccc 60

tctgccatgg tggcacaagt gggacctcag gtggtgaaca tcaatacaaa gctgggctac 120

aacaatgcag tgggagcagg caccggaatc gtgatcgacc caaacggcgt ggtgctgaca 180

aacaatgccg tgatcgccgg cgccaccgac atcaatgcct tttctgtggg cagcggccag 240

acatacggcg tggacgtggt gggctatgat aggacccaga atgtggccgt gctgcagctg 300

aggggagcag gaggcctgcc ttccgccgcc atcggcggcg gcgtggccgt gggcgagcca 360

gtggtggcaa tgggcaacag cggaggacag ggaggcacac cacgggccgt gcccggcaga 420

gtggtggccc tgggacagac cgtgcaggca agcgattccc tgaccggagc cgaggagaca 480

ctgaacggcc tgatccagtt tgatgcagca atccagccag gcgacgcagg aggacctgtg 540

gtgaatggcc tgggacaggt ggtgggaatg aacaccgcag cctctgacaa tttccagctg 600

agccagggag gacagggatt tgcaatccca atcggacagg caatggcaat cgcaggacag 660

atcaggagcg gaggaggctc cccaaccgtg cacatcggac caacagcctt cctgggactg 720

ggagtggtgg acaacaatgg caacggcgcc cgggtgcaga gagtggtggg ctccgcccca 780

gcagcatctc tgggcatcag caccggcgac gtgatcacag ccgtggatgg cgcccctatc 840

aacagcgcca ccgccatggc cgatgccctg aatggccacc acccaggcga cgtgatcagc 900

gtgacctggc agacaaagtc cggaggcacc aggacaggaa acgtgacact ggcagaggga 960

ccaccagca 969

<210> 16

<211> 1044

<212> DNA

<213> artificial sequence

<220>

<223> nucleic acid sequence of PstS3

<400> 16

tgcggcaacg acgataatgt gaccggagga ggagcaacca caggacaggc ctccgccaag 60

gtggattgtg gcggcaagaa gaccctgaag gcatccggct ctacagcaca ggcaaacgcc 120

atgaccaggt tcgtgaacgt gttcgagcag gcatgcccag gacagaccct gaactataca 180

gcaaatggca gcggagcagg catctccgag ttcaacggca atcagacaga ctttggcggc 240

tctgatgtgc ctctgagcaa ggacgagcca tccggagcag caagatgtgg cagccctgcc 300

tggaatctgc cagtggtgtt tggccccatc gccgtgacct acaacctgaa tagcgtgagc 360

tccctgaacc tggatggccc cacactggcc aagattttca acggctccat cacccagtgg 420

aacaatcccg ccatccaggc cctgaaccgg gacttcaccc tgcctggcga gagaatccac 480

gtggtgtttc gctccgacga gtctggcacc acagataact tccagagata cctgcaggca 540

gcatctaatg gagcatgggg caagggagca ggcaagagct ttcagggagg agtgggagag 600

ggagccagag gcaacgatgg caccagcgcc gccgccaaga acaccccagg ctctatcaca 660

tacaatgagt ggagcttcgc ccaggcccac gacctgacca tggccaatat cgtgacaagc 720

gccggaggcg atcctgtggc aatcaccatc gactccgtgg gacagaccat cgcaggagcc 780

acaatctctg gcgtgggcaa cgatctggtg ctggacacag atagcttcta taggcctaag 840

cgcccaggct cctacccaat cgtgctggcc acctatgaga tcgtgtgctc taagtacccc 900

gacagccaag tgggcacagc cgtgaaggcc tttctgcagt ccaccatcgg agcaggacag 960

tctggactgg gcgataacgg ctacatccct atcccagacg agttcaagag cagactgtcc 1020

accgccgtga atgccatcgc ctaa 1044

<210> 17

<211> 1863

<212> DNA

<213> artificial sequence

<220>

<223> nucleic acid sequence of PPE39

<400> 17

gtgaatttca gcgtgctgcc ccctgagatc aactccggcc ggatgttctt tggagcaggc 60

tccggaccta tgctggctgc cgccgccgcc tgggacggac tggcagcaga gctgggcctg 120

gccgccgaga gctttggcct ggtgacaagc ggcctggccg gcggctctgg ccaggcctgg 180

cagggcgctg ccgccgccgc catggtggtg gctgccgccc catacgccgg ctggctggcc 240

gccgccgccg ccagagccgg aggagccgcc gtgcaggcca aggccgtggc cggcgccttc 300

gaggccgcca gggccgcaat ggtggaccca gtggtggtgg cagcaaaccg ctctgccttc 360

gtgcagctgg tgctgagcaa cgtgttcgga cagaacgcac cagcaatcgc tgccgccgag 420

gcaacctacg agcagatgtg ggcagcagat gtggcagcaa tggtgggcta tcacggagga 480

gcaagcgccg ccgccgccgc cctggcccca tggcagcagg ccgtgcccgg cctgagcgga 540

ctgctggact ccgcccagtc ctctgcccag gccgtgaccg cacaggcagt gggctccaca 600

gtgcccggcc ctctgcaggg catcaatttc ggctttggca acatcggctc tctgaatctg 660

ggcagcggca acacaggcga taccaatgtg ggcagcggca acatcggcaa taccaacctg 720

ggcggcggca atatcggctc cttcaacctg ggctctggca atcagggcga catcaacctg 780

ggcatcggca atgtgggcaa tctgaacctg ggcagcggca actttggctc ccagaatctg 840

ggctctggca acatcggcag caccaatgtg ggctccggca acatcggctc tacaaatgtg 900

ggcagcggaa atatcggcga taccaatttc ggcaacggca acaatggcaa tttcaacttt 960

ggctccggca acacaggctc taacaatatc ggctttggca ataccggcag cggcaacttc 1020

ggctttggca atacaggcaa caataacatc ggcatcggcc tgaccggcga cggacagatc 1080

ggaatcggag gcctgaactc tggcagcggc aatatcggct tcggcaactc tggcacaggc 1140

aatgtgggcc tgttcaacag cggcaccggc aatgtgggct ttggcaactc cggcacagcc 1200

aataccggct tcggcaacgc cggcaatgtg aacacaggct tttggaatgg cggctccaca 1260

aacaccggac tggcaaatgc aggagcagga aacaccggct tctttgacgc cggcaattac 1320

aactttggct ctctgaacgc cggcaatatc aacagctcct tcggcaatag cggcgacggc 1380

aactccggct ttctgaatgc cggcgatgtg aactctggcg tgggcaatgc cggcgacgtg 1440

aacacaggcc tgggcaatag cggcaatatc aacaccggcg gcttcaaccc aggcacactg 1500

aataccggct tcttttccgc catgacacag gccggcccca actctggctt ctttaatgca 1560

ggcaccggca acagcggatt tggacacaac gacccagcag gcagcggcaa ttccggcatc 1620

cagaactctg gcttcggcaa tagcggctac gtgaacacat ctaccacaag catgtttggc 1680

ggcaattctg gcgtgctgaa caccggctac ggcaatagcg gcttctataa cgccgccgtg 1740

aataacacag gcatctttgt gaccggcgtg atgtctagcg gcttctttaa tttcggcaca 1800

ggcaactccg gcctgctggt gtctggcaat ggcctgagcg gcttctttaa gaacctgttt 1860

gga 1863

<210> 18

<211> 2220

<212> DNA

<213> artificial sequence

<220>

<223> nucleic acid sequence of GlcB

<400> 18

accgaccgcg tgtccgtggg caacctgaga atcgccaggg tgctgtatga tttcgtgaat 60

aacgaggccc tgcctggcac agacatcgac ccagacagct tttgggcagg agtggataag 120

gtggtggccg acctgacccc tcagaatcag gccctgctga acgccagaga cgagctgcag 180

gcccagatcg acaagtggca ccggcggaga gtgatcgagc caatcgacat ggacgcctac 240

aggcagttcc tgacagagat cggctatctg ctgccagagc ccgacgactt caccatcacc 300

acatccggcg tggatgccga gatcaccaca accgcaggac ctcagctggt ggtgccagtg 360

ctgaacgcaa ggttcgccct gaatgcagca aacgccagat ggggcagcct gtacgatgcc 420

ctgtatggca cagacgtgat cccagagacc gatggagcag agaagggacc tacatacaat 480

aaggtgaggg gcgacaaagt gatcgcctat gcccgcaagt tcctggacga ttctgtgccc 540

ctgtcctctg gcagcttcgg cgacgccaca ggctttaccg tgcaggatgg acagctggtg 600

gtggccctgc ctgataagtc caccggactg gcaaacccag gacagtttgc aggatacacc 660

ggagcagcag agtcccccac atctgtgctg ctgatcaatc acggcctgca catcgagatc 720

ctgatcgacc ctgagagcca agtgggcaca accgacagag caggagtgaa ggatgtgatc 780

ctggagtccg ccatcacaac catcatggat ttcgaggact ctgtggcagc agtggatgca 840

gcagacaagg tgctgggcta cagaaattgg ctgggcctga acaagggcga tctggctgcc 900

gccgtggata aggacggcac cgcctttctg agggtgctga atagagacag gaactatacc 960

gccccaggag gaggacagtt cacactgcca ggccggtctc tgatgtttgt gagaaatgtg 1020

ggccacctga tgaccaacga cgccatcgtg gatacagacg gcagcgaggt gttcgagggc 1080

atcatggatg ccctgtttac aggcctgatc gccatccacg gcctgaaggc ctccgacgtg 1140

aatggccccc tgatcaactc ccgcaccggc tctatctaca tcgtgaagcc aaagatgcac 1200

ggacctgcag aggtggcctt cacatgcgag ctgtttagcc gggtggagga cgtgctgggc 1260

ctgcctcaga ataccatgaa gatcggcatc atggatgagg agcgccggac aaccgtgaac 1320

ctgaaggcct gtatcaaggc tgccgccgac agggtggtgt tcatcaatac aggctttctg 1380

gatcgcaccg gcgacgagat ccacacatcc atggaggccg gcccaatggt gagaaagggc 1440

accatgaagt ctcagccctg gattctggcc tatgaggatc acaacgtgga cgcaggactg 1500

gcagcaggct tcagcggcag agcccaagtg ggcaagggca tgtggacaat gaccgagctg 1560

atggccgata tggtggagac aaagatcgca cagcctaggg ccggagccag caccgcatgg 1620

gtgccttccc caacagcagc caccctgcac gcactgcact accaccaggt ggacgtggca 1680

gcagtgcagc agggactggc aggcaagaga agggccacca tcgagcagct gctgacaatc 1740

cctctggcaa aggagctggc atgggcacca gacgagatca gggaggaggt ggataataac 1800

tgccagtcca tcctgggcta tgtggtgcgg tgggtggatc agggagtggg atgttctaag 1860

gtgccagaca tccacgacgt ggccctgatg gaggacaggg ccacactgcg gatcagctcc 1920

cagctgctgg ccaactggct gaggcacggc gtgatcacct ctgccgacgt gcgcgcaagc 1980

ctggagagga tggcaccact ggtggacagg cagaatgcag gcgatgtggc ctacagacct 2040

atggccccaa acttcgacga tagcatcgcc tttctggcag cacaggagct gatcctgtcc 2100

ggagcacagc agccaaacgg atataccgag cctatcctgc accgccggag aagggagttc 2160

aaggccagag ccgcagagaa gccagcacct tccgataggg caggcgacga tgcagcccgc 2220

2220

<210> 19

<211> 1302

<212> DNA

<213> artificial sequence

<220>

<223> nucleic acid sequence of RipA

<400> 19

gatccccaga cagacaccat cgccgccctg atcgcagacg tggcaaaggc caaccagagg 60

ctgcaggatc tgagcgacga ggtgcaggca gagcaggagt ccgtgaataa ggcaatggtg 120

gacgtggaga ccgcccggga taacgctgcc gccgccgagg acgatctgga ggtgagccag 180

agagccgtga aggatgcaaa cgcagcaatc gctgccgccc agcacaggtt cgacaccttc 240

gccgccgcca cctacatgaa tggcccctct gtgagctatc tgagcgcctc tagccctgac 300

gagatcatcg ccacagtgac cgcagcaaag accctgtccg cctcctctca ggccgtgatg 360

gcaaatctgc agagggcacg gacagagagg gtgaacaccg agtccgccgc ccgcctggca 420

aagcagaagg cagataaggc tgccgccgac gcaaaggcat ctcaggatgc agcagtggcc 480

gccctgacag agaccaggcg gaagttcgac gagcagagag aggaggtgca gaggctggca 540

gcagagaggg atgcagcaca ggcccggctg caggccgcca gactggtggc atggagctcc 600

gagggaggac agggcgcccc accctttcgg atgtgggacc ccggcagcgg accagcaggc 660

ggcagagcct gggatggact gtgggacccc accctgccta tgatcccatc cgccaatatc 720

cccggcgacc ctatcgccgt ggtgaaccag gtgctgggca tcagcgccac atccgcccag 780

gtgacagcca acatgggcag aaagttcctg gagcagctgg gcatcctgca gcccacagac 840

accggaatca ccaatgcacc tgcaggcagc gcccagggac gcatcccacg ggtgtacggc 900

aggcaggcat ccgagtatgt gatcagaagg ggcatgtccc agatcggcgt gccttactct 960

tggggaggag gaaacgcagc aggaccatcc aagggaatcg actctggagc aggcaccgtg 1020

ggattcgatt gttccggcct ggtgctgtat tcttttgccg gcgtgggcat caagctgcca 1080

cactactccg gctctcagta taatctgggc cgcaagatcc cctctagcca gatgcgccgg 1140

ggcgatgtga tcttctacgg ccctaatggc agccagcacg tgaccatcta tctgggaaac 1200

ggacagatgc tggaggcacc agacgtgggc ctgaaggtga gggtggcacc tgtgcgcaca 1260

gcaggcatga ccccatacgt ggtgcggtac atcgagtatt aa 1302

<210> 20

<211> 30

<212> DNA

<213> artificial sequence

<220>

<223> nucleic acid sequence of GS linker

<400> 20

ggcagcggct ccggctctgg cagcggctct 30

<210> 21

<211> 30

<212> DNA

<213> artificial sequence

<220>

<223> nucleic acid sequence of GS linker

<400> 21

ggctccggct ctggcagcgg ctccggctct 30

<210> 22

<211> 30

<212> DNA

<213> artificial sequence

<220>

<223> nucleic acid sequence of GS linker

<400> 22

ggctctggca gcggctccgg ctctggcagc 30

<210> 23

<211> 30

<212> DNA

<213> artificial sequence

<220>

<223> nucleic acid sequence of GS linker

<400> 23

ggcagcggct ccggctctgg cagcggctcc 30

<210> 24

<211> 69

<212> DNA

<213> artificial sequence

<220>

<223> nucleic acid sequence of tPA

<400> 24

atggatgcca tgaagagggg actgtgctgc gtgctgctgc tgtgcggagc cgtgttcgtg 60

tctccaagc 69

<210> 25

<211> 69

<212> DNA

<213> artificial sequence

<220>

<223> nucleic acid sequence of tPA

<400> 25

atggatgcca tgaagagggg actgtgctgc gtgctgctgc tgtgcggagc cgtgttcgtg 60

tccccatct 69

<210> 26

<211> 474

<212> DNA

<213> artificial sequence

<220>

<223> Flt3L nucleic acid sequence

<400> 26

acccaggatt gttctttcca gcacagcccc atcagctccg actttgccgt gaagatccgc 60

gagctgagcg actacctgct gcaggattat cccgtgacag tggcctccaa tctgcaggac 120

gaggagctgt gcggaggact gtggaggctg gtgctggcac agcggtggat ggagagactg 180

aagaccgtgg caggctccaa gatgcaggga ctgctggagc gcgtgaacac cgagatccac 240

ttcgtgacaa agtgcgcctt tcagccccct ccaagctgtc tgcggtttgt gcagacaaac 300

atctccagac tgctgcagga gacctctgag cagctggtgg ccctgaagcc ctggatcaca 360

aggcagaatt tctctcgctg cctggagctg cagtgtcagc ctgactctag caccctgcca 420

cctccatgga gccctcggcc actggaggca accgcaccaa cagcacctgg ctcc 474

<210> 27

<211> 474

<212> DNA

<213> artificial sequence

<220>

<223> Flt3L nucleic acid sequence

<400> 27

cacgccaccc aggattgttc tttccagcac agccctatca gctccgactt tgccgtgaag 60

atccgggagc tgtctgatta cctgctgcag gactatcccg tgacagtggc cagcaacctg 120

caggacgagg agctgtgcgg aggactgtgg aggctggtgc tggcacagag atggatggag 180

aggctgaaga ccgtggcagg ctccaagatg cagggactgc tggagcgggt gaatacagag 240

atccacttcg tgaccaagtg cgcctttcag ccccctccaa gctgtctgcg gttcgtgcag 300

accaacatca gcagactgct gcaggagaca tccgagcagc tggtggccct gaagccttgg 360

atcacaagac agaatttttc taggtgcctg gagctgcagt gtcagccaga ttctagcacc 420

ctgccacctc catggagccc tcgcccactg gaggccacag caccaaccgc acca 474

<210> 28

<211> 3846

<212> DNA

<213> artificial sequence

<220>

<223> nucleic acid sequence of GX-170A

<400> 28

atggatgcca tgaagagggg actgtgctgc gtgctgctgc tgtgcggagc cgtgttcgtg 60

tctccaagcc acgccaccca ggattgttct ttccagcaca gccccatcag ctccgacttt 120

gccgtgaaga tccgcgagct gagcgactac ctgctgcagg attatcccgt gacagtggcc 180

tccaatctgc aggacgagga gctgtgcgga ggactgtgga ggctggtgct ggcacagcgg 240

tggatggaga gactgaagac cgtggcaggc tccaagatgc agggactgct ggagcgcgtg 300

aacaccgaga tccacttcgt gacaaagtgc gcctttcagc cccctccaag ctgtctgcgg 360

tttgtgcaga caaacatctc cagactgctg caggagacct ctgagcagct ggtggccctg 420

aagccctgga tcacaaggca gaatttctct cgctgcctgg agctgcagtg tcagcctgac 480

tctagcaccc tgccacctcc atggagccct cggccactgg aggcaaccgc accaacagca 540

cctggctccg gctctggcag cggctccggc tcttttagca ggccaggcct gcccgtggag 600

tatctgcagg tgccctcccc ttctatgggc cgcgacatca aggtgcagtt ccagtccgga 660

ggagccaact ctccagccct gtatctgctg gacggcctga gagcccagga cgatttcagc 720

ggctgggaca tcaatacacc cgcctttgag tggtacgacc agagcggact gtccgtggtc 780

atgccagtgg gaggacagtc ctctttttac tccgactggt atcagcctgc atgcggcaag 840

gcaggatgtc agacctacaa gtgggagacc ttcctgacaa gcgagctgcc tggctggctg 900

caggccaaca ggcacgtgaa gccaacaggc tccgccgtgg tgggactgtc tatggcagcc 960

agctccgccc tgaccctggc aatctaccac cctcagcagt tcgtgtacgc aggagcaatg 1020

agcggactgc tggacccctc ccaggcaatg ggaccaaccc tgatcggact ggcaatgggc 1080

gacgcaggag gatataaggc aagcgacatg tggggaccaa aggaggaccc cgcatggcag 1140

aggaacgacc cactgctgaa tgtgggcaag ctgatcgcca acaataccag agtgtgggtg 1200

tactgtggaa acggcaagcc atccgatctg ggaggaaaca atctgcctgc caagttcctg 1260

gagggctttg tgcggacatc taatatcaag ttccaggacg catataacgc aggaggagga 1320

cacaatggcg tgttcgactt tcccgattcc ggcacccact cttgggagta ctggggagca 1380

cagctgaacg caatgaagcc cgacctgcag agagccctgg gagcaacccc caatacagga 1440

ccagcaccac agggagcagg cagcggctcc ggctctggca gcggctctag ccagatcatg 1500

tacaactatc ctgccatgct gggacacgca ggcgacatgg caggatacgc aggcaccctg 1560

cagtctctgg gagcagagat cgcagtggag caggccgccc tgcagagcgc ctggcagggc 1620

gataccggca tcacatatca ggcatggcag gcacagtgga accaggcaat ggaggacctg 1680

gtgcgggcct accacgccat gtcctctacc cacgaggcca atacaatggc catgatggcc 1740

agagataccg cagaggcagc aaagtgggga ggaggctccg gctctggcag cggctccggc 1800

tctgcccccc ctgccctgtc ccaggaccgg ttcgccgatt ttcctgccct gccactggac 1860

ccctctgcca tggtggcaca agtgggacct caggtggtga acatcaatac aaagctgggc 1920

tacaacaatg cagtgggagc aggcaccgga atcgtgatcg acccaaacgg cgtggtgctg 1980

acaaacaatg ccgtgatcgc cggcgccacc gacatcaatg ccttttctgt gggcagcggc 2040

cagacatacg gcgtggacgt ggtgggctat gataggaccc agaatgtggc cgtgctgcag 2100

ctgaggggag caggaggcct gccttccgcc gccatcggcg gcggcgtggc cgtgggcgag 2160

ccagtggtgg caatgggcaa cagcggagga cagggaggca caccacgggc cgtgcccggc 2220

agagtggtgg ccctgggaca gaccgtgcag gcaagcgatt ccctgaccgg agccgaggag 2280

acactgaacg gcctgatcca gtttgatgca gcaatccagc caggcgacgc aggaggacct 2340

gtggtgaatg gcctgggaca ggtggtggga atgaacaccg cagcctctga caatttccag 2400

ctgagccagg gaggacaggg atttgcaatc ccaatcggac aggcaatggc aatcgcagga 2460

cagatcagga gcggaggagg ctccccaacc gtgcacatcg gaccaacagc cttcctggga 2520

ctgggagtgg tggacaacaa tggcaacggc gcccgggtgc agagagtggt gggctccgcc 2580

ccagcagcat ctctgggcat cagcaccggc gacgtgatca cagccgtgga tggcgcccct 2640

atcaacagcg ccaccgccat ggccgatgcc ctgaatggcc accacccagg cgacgtgatc 2700

agcgtgacct ggcagacaaa gtccggaggc accaggacag gaaacgtgac actggcagag 2760

ggaccaccag caggctctgg cagcggctcc ggctctggca gctgcggcaa cgacgataat 2820

gtgaccggag gaggagcaac cacaggacag gcctccgcca aggtggattg tggcggcaag 2880

aagaccctga aggcatccgg ctctacagca caggcaaacg ccatgaccag gttcgtgaac 2940

gtgttcgagc aggcatgccc aggacagacc ctgaactata cagcaaatgg cagcggagca 3000

ggcatctccg agttcaacgg caatcagaca gactttggcg gctctgatgt gcctctgagc 3060

aaggacgagc catccggagc agcaagatgt ggcagccctg cctggaatct gccagtggtg 3120

tttggcccca tcgccgtgac ctacaacctg aatagcgtga gctccctgaa cctggatggc 3180

cccacactgg ccaagatttt caacggctcc atcacccagt ggaacaatcc cgccatccag 3240

gccctgaacc gggacttcac cctgcctggc gagagaatcc acgtggtgtt tcgctccgac 3300

gagtctggca ccacagataa cttccagaga tacctgcagg cagcatctaa tggagcatgg 3360

ggcaagggag caggcaagag ctttcaggga ggagtgggag agggagccag aggcaacgat 3420

ggcaccagcg ccgccgccaa gaacacccca ggctctatca catacaatga gtggagcttc 3480

gcccaggccc acgacctgac catggccaat atcgtgacaa gcgccggagg cgatcctgtg 3540

gcaatcacca tcgactccgt gggacagacc atcgcaggag ccacaatctc tggcgtgggc 3600

aacgatctgg tgctggacac agatagcttc tataggccta agcgcccagg ctcctaccca 3660

atcgtgctgg ccacctatga gatcgtgtgc tctaagtacc ccgacagcca agtgggcaca 3720

gccgtgaagg cctttctgca gtccaccatc ggagcaggac agtctggact gggcgataac 3780

ggctacatcc ctatcccaga cgagttcaag agcagactgt ccaccgccgt gaatgccatc 3840

gcctaa 3846

<210> 29

<211> 6018

<212> DNA

<213> artificial sequence

<220>

<223> nucleic acid sequence of GX-170B

<400> 29

atggatgcca tgaagagggg actgtgctgc gtgctgctgc tgtgcggagc cgtgttcgtg 60

tccccatctc acgccaccca ggattgttct ttccagcaca gccctatcag ctccgacttt 120

gccgtgaaga tccgggagct gtctgattac ctgctgcagg actatcccgt gacagtggcc 180

agcaacctgc aggacgagga gctgtgcgga ggactgtgga ggctggtgct ggcacagaga 240

tggatggaga ggctgaagac cgtggcaggc tccaagatgc agggactgct ggagcgggtg 300

aatacagaga tccacttcgt gaccaagtgc gcctttcagc cccctccaag ctgtctgcgg 360

ttcgtgcaga ccaacatcag cagactgctg caggagacat ccgagcagct ggtggccctg 420

aagccttgga tcacaagaca gaatttttct aggtgcctgg agctgcagtg tcagccagat 480

tctagcaccc tgccacctcc atggagccct cgcccactgg aggccacagc accaaccgca 540

ccaggcagcg gctccggctc tggcagcggc tccgtgaatt tcagcgtgct gccccctgag 600

atcaactccg gccggatgtt ctttggagca ggctccggac ctatgctggc tgccgccgcc 660

gcctgggacg gactggcagc agagctgggc ctggccgccg agagctttgg cctggtgaca 720

agcggcctgg ccggcggctc tggccaggcc tggcagggcg ctgccgccgc cgccatggtg 780

gtggctgccg ccccatacgc cggctggctg gccgccgccg ccgccagagc cggaggagcc 840

gccgtgcagg ccaaggccgt ggccggcgcc ttcgaggccg ccagggccgc aatggtggac 900

ccagtggtgg tggcagcaaa ccgctctgcc ttcgtgcagc tggtgctgag caacgtgttc 960

ggacagaacg caccagcaat cgctgccgcc gaggcaacct acgagcagat gtgggcagca 1020

gatgtggcag caatggtggg ctatcacgga ggagcaagcg ccgccgccgc cgccctggcc 1080

ccatggcagc aggccgtgcc cggcctgagc ggactgctgg actccgccca gtcctctgcc 1140

caggccgtga ccgcacaggc agtgggctcc acagtgcccg gccctctgca gggcatcaat 1200

ttcggctttg gcaacatcgg ctctctgaat ctgggcagcg gcaacacagg cgataccaat 1260

gtgggcagcg gcaacatcgg caataccaac ctgggcggcg gcaatatcgg ctccttcaac 1320

ctgggctctg gcaatcaggg cgacatcaac ctgggcatcg gcaatgtggg caatctgaac 1380

ctgggcagcg gcaactttgg ctcccagaat ctgggctctg gcaacatcgg cagcaccaat 1440

gtgggctccg gcaacatcgg ctctacaaat gtgggcagcg gaaatatcgg cgataccaat 1500

ttcggcaacg gcaacaatgg caatttcaac tttggctccg gcaacacagg ctctaacaat 1560

atcggctttg gcaataccgg cagcggcaac ttcggctttg gcaatacagg caacaataac 1620

atcggcatcg gcctgaccgg cgacggacag atcggaatcg gaggcctgaa ctctggcagc 1680

ggcaatatcg gcttcggcaa ctctggcaca ggcaatgtgg gcctgttcaa cagcggcacc 1740

ggcaatgtgg gctttggcaa ctccggcaca gccaataccg gcttcggcaa cgccggcaat 1800

gtgaacacag gcttttggaa tggcggctcc acaaacaccg gactggcaaa tgcaggagca 1860

ggaaacaccg gcttctttga cgccggcaat tacaactttg gctctctgaa cgccggcaat 1920

atcaacagct ccttcggcaa tagcggcgac ggcaactccg gctttctgaa tgccggcgat 1980

gtgaactctg gcgtgggcaa tgccggcgac gtgaacacag gcctgggcaa tagcggcaat 2040

atcaacaccg gcggcttcaa cccaggcaca ctgaataccg gcttcttttc cgccatgaca 2100

caggccggcc ccaactctgg cttctttaat gcaggcaccg gcaacagcgg atttggacac 2160

aacgacccag caggcagcgg caattccggc atccagaact ctggcttcgg caatagcggc 2220

tacgtgaaca catctaccac aagcatgttt ggcggcaatt ctggcgtgct gaacaccggc 2280

tacggcaata gcggcttcta taacgccgcc gtgaataaca caggcatctt tgtgaccggc 2340

gtgatgtcta gcggcttctt taatttcggc acaggcaact ccggcctgct ggtgtctggc 2400

aatggcctga gcggcttctt taagaacctg tttggaggct ccggctctgg cagcggctcc 2460

ggctctaccg accgcgtgtc cgtgggcaac ctgagaatcg ccagggtgct gtatgatttc 2520

gtgaataacg aggccctgcc tggcacagac atcgacccag acagcttttg ggcaggagtg 2580

gataaggtgg tggccgacct gacccctcag aatcaggccc tgctgaacgc cagagacgag 2640

ctgcaggccc agatcgacaa gtggcaccgg cggagagtga tcgagccaat cgacatggac 2700

gcctacaggc agttcctgac agagatcggc tatctgctgc cagagcccga cgacttcacc 2760

atcaccacat ccggcgtgga tgccgagatc accacaaccg caggacctca gctggtggtg 2820

ccagtgctga acgcaaggtt cgccctgaat gcagcaaacg ccagatgggg cagcctgtac 2880

gatgccctgt atggcacaga cgtgatccca gagaccgatg gagcagagaa gggacctaca 2940

tacaataagg tgaggggcga caaagtgatc gcctatgccc gcaagttcct ggacgattct 3000

gtgcccctgt cctctggcag cttcggcgac gccacaggct ttaccgtgca ggatggacag 3060

ctggtggtgg ccctgcctga taagtccacc ggactggcaa acccaggaca gtttgcagga 3120

tacaccggag cagcagagtc ccccacatct gtgctgctga tcaatcacgg cctgcacatc 3180

gagatcctga tcgaccctga gagccaagtg ggcacaaccg acagagcagg agtgaaggat 3240

gtgatcctgg agtccgccat cacaaccatc atggatttcg aggactctgt ggcagcagtg 3300

gatgcagcag acaaggtgct gggctacaga aattggctgg gcctgaacaa gggcgatctg 3360

gctgccgccg tggataagga cggcaccgcc tttctgaggg tgctgaatag agacaggaac 3420

tataccgccc caggaggagg acagttcaca ctgccaggcc ggtctctgat gtttgtgaga 3480

aatgtgggcc acctgatgac caacgacgcc atcgtggata cagacggcag cgaggtgttc 3540

gagggcatca tggatgccct gtttacaggc ctgatcgcca tccacggcct gaaggcctcc 3600

gacgtgaatg gccccctgat caactcccgc accggctcta tctacatcgt gaagccaaag 3660

atgcacggac ctgcagaggt ggccttcaca tgcgagctgt ttagccgggt ggaggacgtg 3720

ctgggcctgc ctcagaatac catgaagatc ggcatcatgg atgaggagcg ccggacaacc 3780

gtgaacctga aggcctgtat caaggctgcc gccgacaggg tggtgttcat caatacaggc 3840

tttctggatc gcaccggcga cgagatccac acatccatgg aggccggccc aatggtgaga 3900

aagggcacca tgaagtctca gccctggatt ctggcctatg aggatcacaa cgtggacgca 3960

ggactggcag caggcttcag cggcagagcc caagtgggca agggcatgtg gacaatgacc 4020

gagctgatgg ccgatatggt ggagacaaag atcgcacagc ctagggccgg agccagcacc 4080

gcatgggtgc cttccccaac agcagccacc ctgcacgcac tgcactacca ccaggtggac 4140

gtggcagcag tgcagcaggg actggcaggc aagagaaggg ccaccatcga gcagctgctg 4200

acaatccctc tggcaaagga gctggcatgg gcaccagacg agatcaggga ggaggtggat 4260

aataactgcc agtccatcct gggctatgtg gtgcggtggg tggatcaggg agtgggatgt 4320

tctaaggtgc cagacatcca cgacgtggcc ctgatggagg acagggccac actgcggatc 4380

agctcccagc tgctggccaa ctggctgagg cacggcgtga tcacctctgc cgacgtgcgc 4440

gcaagcctgg agaggatggc accactggtg gacaggcaga atgcaggcga tgtggcctac 4500

agacctatgg ccccaaactt cgacgatagc atcgcctttc tggcagcaca ggagctgatc 4560

ctgtccggag cacagcagcc aaacggatat accgagccta tcctgcaccg ccggagaagg 4620

gagttcaagg ccagagccgc agagaagcca gcaccttccg atagggcagg cgacgatgca 4680

gcccgcggca gcggctccgg ctctggcagc ggctccgatc cccagacaga caccatcgcc 4740

gccctgatcg cagacgtggc aaaggccaac cagaggctgc aggatctgag cgacgaggtg 4800

caggcagagc aggagtccgt gaataaggca atggtggacg tggagaccgc ccgggataac 4860

gctgccgccg ccgaggacga tctggaggtg agccagagag ccgtgaagga tgcaaacgca 4920

gcaatcgctg ccgcccagca caggttcgac accttcgccg ccgccaccta catgaatggc 4980

ccctctgtga gctatctgag cgcctctagc cctgacgaga tcatcgccac agtgaccgca 5040

gcaaagaccc tgtccgcctc ctctcaggcc gtgatggcaa atctgcagag ggcacggaca 5100

gagagggtga acaccgagtc cgccgcccgc ctggcaaagc agaaggcaga taaggctgcc 5160

gccgacgcaa aggcatctca ggatgcagca gtggccgccc tgacagagac caggcggaag 5220

ttcgacgagc agagagagga ggtgcagagg ctggcagcag agagggatgc agcacaggcc 5280

cggctgcagg ccgccagact ggtggcatgg agctccgagg gaggacaggg cgccccaccc 5340

tttcggatgt gggaccccgg cagcggacca gcaggcggca gagcctggga tggactgtgg 5400

gaccccaccc tgcctatgat cccatccgcc aatatccccg gcgaccctat cgccgtggtg 5460

aaccaggtgc tgggcatcag cgccacatcc gcccaggtga cagccaacat gggcagaaag 5520

ttcctggagc agctgggcat cctgcagccc acagacaccg gaatcaccaa tgcacctgca 5580

ggcagcgccc agggacgcat cccacgggtg tacggcaggc aggcatccga gtatgtgatc 5640

agaaggggca tgtcccagat cggcgtgcct tactcttggg gaggaggaaa cgcagcagga 5700

ccatccaagg gaatcgactc tggagcaggc accgtgggat tcgattgttc cggcctggtg 5760

ctgtattctt ttgccggcgt gggcatcaag ctgccacact actccggctc tcagtataat 5820

ctgggccgca agatcccctc tagccagatg cgccggggcg atgtgatctt ctacggccct 5880

aatggcagcc agcacgtgac catctatctg ggaaacggac agatgctgga ggcaccagac 5940

gtgggcctga aggtgagggt ggcacctgtg cgcacagcag gcatgacccc atacgtggtg 6000

cggtacatcg agtattaa 6018

<210> 30

<211> 253

<212> PRT

<213> artificial sequence

<220>

<223> amino acid sequence of IL-12p35

<400> 30

Met Trp Pro Pro Gly Ser Ala Ser Gln Pro Pro Pro Ser Pro Ala Ala

1 5 10 15

Ala Thr Gly Leu His Pro Ala Ala Arg Pro Val Ser Leu Gln Cys Arg

20 25 30

Leu Ser Met Cys Pro Ala Arg Ser Leu Leu Leu Val Ala Thr Leu Val

35 40 45

Leu Leu Asp His Leu Ser Leu Ala Arg Asn Leu Pro Val Ala Thr Pro

50 55 60

Asp Pro Gly Met Phe Pro Cys Leu His His Ser Gln Asn Leu Leu Arg

65 70 75 80

Ala Val Ser Asn Met Leu Gln Lys Ala Arg Gln Thr Leu Glu Phe Tyr

85 90 95

Pro Cys Thr Ser Glu Glu Ile Asp His Glu Asp Ile Thr Lys Asp Lys

100 105 110

Thr Ser Thr Val Glu Ala Cys Leu Pro Leu Glu Leu Thr Lys Asn Glu

115 120 125

Ser Cys Leu Asn Ser Arg Glu Thr Ser Phe Ile Thr Asn Gly Ser Cys

130 135 140

Leu Ala Ser Arg Lys Thr Ser Phe Met Met Ala Leu Cys Leu Ser Ser

145 150 155 160

Ile Tyr Glu Asp Leu Lys Met Tyr Gln Val Glu Phe Lys Thr Met Asn

165 170 175

Ala Lys Leu Leu Met Asp Pro Lys Arg Gln Ile Phe Leu Asp Gln Asn

180 185 190

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

195 200 205

Glu Thr Val Pro Gln Lys Ser Ser Leu Glu Glu Pro Asp Phe Tyr Lys

210 215 220

Thr Lys Ile Lys Leu Cys Ile Leu Leu His Ala Phe Arg Ile Arg Ala

225 230 235 240

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

245 250

<210> 31

<211> 328

<212> PRT

<213> artificial sequence

<220>

<223> amino acid sequence of IL-12p40

<400> 31

Met Cys His Gln Gln Leu Val Ile Ser Trp Phe Ser Leu Val Phe Leu

1 5 10 15

Ala Ser Pro Leu Val Ala Ile Trp Glu Leu Lys Lys Asp Val Tyr Val

20 25 30

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

35 40 45

Thr Cys Asp Thr Pro Glu Glu Asp Gly Ile Thr Trp Thr Leu Asp Gln

50 55 60

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

65 70 75 80

Glu Phe Gly Asp Ala Gly Gln Tyr Thr Cys His Lys Gly Gly Glu Val

85 90 95

Leu Ser His Ser Leu Leu Leu Leu His Lys Lys Glu Asp Gly Ile Trp

100 105 110

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

115 120 125

Leu Arg Cys Glu Ala Lys Asn Tyr Ser Gly Arg Phe Thr Cys Trp Trp

130 135 140

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

145 150 155 160

Gly Ser Ser Asp Pro Gln Gly Val Thr Cys Gly Ala Ala Thr Leu Ser

165 170 175

Ala Glu Arg Val Arg Gly Asp Asn Lys Glu Tyr Glu Tyr Ser Val Glu

180 185 190

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

195 200 205

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

210 215 220

Ser Ser Phe Phe Ile Arg Asp Ile Ile Lys Pro Asp Pro Pro Lys Asn

225 230 235 240

Leu Gln Leu Lys Pro Leu Lys Asn Ser Arg Gln Val Glu Val Ser Trp

245 250 255

Glu Tyr Pro Asp Thr Trp Ser Thr Pro His Ser Tyr Phe Ser Leu Thr

260 265 270

Phe Cys Val Gln Val Gln Gly Lys Ser Lys Arg Glu Lys Lys Asp Arg

275 280 285

Val Phe Thr Asp Lys Thr Ser Ala Thr Val Ile Cys Arg Lys Asn Ala

290 295 300

Ser Ile Ser Val Arg Ala Gln Asp Arg Tyr Tyr Ser Ser Ser Trp Ser

305 310 315 320

Glu Trp Ala Ser Val Pro Cys Ser

325

<210> 32

<211> 155

<212> PRT

<213> artificial sequence

<220>

<223> amino acid sequence of IL-21

<400> 32

Met Glu Arg Ile Val Ile Cys Leu Met Val Ile Phe Leu Gly Thr Leu

1 5 10 15

Val His Lys Ser Ser Ser Gln Gly Gln Asp Arg His Met Ile Arg Met

20 25 30

Arg Gln Leu Ile Asp Ile Val Asp Gln Leu Lys Asn Tyr Val Asn Asp

35 40 45

Leu Val Pro Glu Phe Leu Pro Ala Pro Glu Asp Val Glu Thr Asn Cys

50 55 60

Glu Trp Ser Ala Phe Ser Cys Phe Gln Lys Ala Gln Leu Lys Ser Ala

65 70 75 80

Asn Thr Gly Asn Asn Glu Arg Ile Ile Asn Val Ser Ile Lys Lys Leu

85 90 95

Lys Arg Lys Pro Pro Ser Thr Asn Ala Gly Arg Arg Gln Lys His Arg

100 105 110

Leu Thr Cys Pro Ser Cys Asp Ser Tyr Glu Lys Lys Pro Pro Lys Glu

115 120 125

Phe Leu Glu Arg Phe Lys Ser Leu Leu Gln Lys Met Ile His Gln His

130 135 140

Leu Ser Ser Arg Thr His Gly Ser Glu Asp Ser

145 150 155

<210> 33

<211> 92

<212> PRT

<213> artificial sequence

<220>

<223> amino acid sequence of MIP-1alpha

<400> 33

Met Gln Val Ser Thr Ala Ala Leu Ala Val Leu Leu Cys Thr Met Ala

1 5 10 15

Leu Cys Asn Gln Phe Ser Ala Ser Leu Ala Ala Asp Thr Pro Thr Ala

20 25 30

Cys Cys Phe Ser Tyr Thr Ser Arg Gln Ile Pro Gln Asn Phe Ile Ala

35 40 45

Asp Tyr Phe Glu Thr Ser Ser Gln Cys Ser Lys Pro Gly Val Ile Phe

50 55 60

Leu Thr Lys Arg Ser Arg Gln Val Cys Ala Asp Pro Ser Glu Glu Trp

65 70 75 80

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

85 90

<210> 34

<211> 712

<212> DNA

<213> artificial sequence

<220>

<223> nucleic acid sequence of IL-12p35

<400> 34

atgtggcccc ctgggtcagc ctcccagcca ccgccctcac ctgccgcggc cacaggtctg 60

catccagcgg ctcgccctgt gtccctgcag tgccggctca gcatgtgtcc agcgcgcagc 120

ctcctccttg tggctaccct ggtcctcctg aatgttccca tgccttcacc actcccaaaa 180

cctgctgagg gccgtcagca acatgctcca gaaggccaga caaactctgg aattttaccc 240

ttgcacttct gaagagattg atcatgaaga tatcacaaaa gataaaacga gcacagtgga 300

ggcctgttta ccattggaat taaccaagaa tgagagttgc ctaaattcca gagagacctc 360

tttcataact aatgggagtt gcctggcctc cagaaagacc tcttttatga tggccctgtg 420

ccttagtagt atttatgaag acttgaagat gtaccaggtg gagttcaaga ccatgaatgc 480

aaagcttctg atggatccta agaggcagat ctttctggat caaaacatgc tggcagttat 540

tgatgagctg atgcaggccc tgaatttcaa cagtgagact gtgccacaaa aatcctccct 600

tgaagaaccg gatttttata aaactaaaat caagctctgc atacttcttc atgctttcag 660

gattcgggca gtgactattg atagagtgat gagctatctg aatgcttcct aa 712

<210> 35

<211> 937

<212> DNA

<213> artificial sequence

<220>

<223> nucleic acid sequence of IL-12p40

<400> 35

atgtgccacc agcagctggt catcagctgg ttctccctgg tctttctggc ttctcctctg 60

gtggcaattt gggagctgaa gaaagacgtg tacgtggtcg aactggactg gtatccagat 120

gcccccggag agatggtggt cctgacctgc gacacaccag aggaagatgg catcacttgg 180

accctggacc agagctccga ggtcctggga agcggcaaga cactgactat tcaggtgaaa 240

gaattcgggg atgctggaca gtacacatgt cataagggcg gggaggtgct gtcccactct 300

ctgctgctgc tgcataagaa agaagatggc atctggtcta ctgacattct gaaggatcag 360

aaagagccca agaacaaaac cttcctgaga tgcgaagcca agaattatag cgggaggttt 420

acctgttggt ggctgaccac aatctctact agtgacctgc ggagcagcta cactgagcgc 480

agagcgggtc agaggagaca acaaggagta cgaatattcc gtggagtgcc aggaagattc 540

tgcatgtccc gcagccgagg aatccctgcc tatcgaagtg atggtggacg ccgtgcacaa 600

gctgaaatac gaaaactaca catcctcttt ctttatccgg gacatcatta agccagatcc 660

ccctaaaaac ctgcagctga agcccctgaa aaattcacga caggtggagg tcagctggga 720

ataccctgat acatggagca ctccacattc ttatttcagt ctgacttttt gcgtgcaggt 780

ccagggcaag agtaaacgag agaagaaaga ccgggtcttc accgataaga catccgctac 840

tgtgatctgt cggaaaaacg ccagtatttc agtgagggct caggaccgct actatagttc 900

aagctggtca gagtgggcaa gcgtgccctg ttcctag 937

<210> 36

<211> 468

<212> DNA

<213> artificial sequence

<220>

<223> nucleic acid sequence of IL-21

<400> 36

atggaacgga ttgtcatttg cctgatggtc atttttctgg gaaccctggt ccacaagtca 60

agcagtcagg gccaggatag gcacatgatt aggatgcgcc agctgatcga cattgtggat 120

cagctgaaga actacgtgaa tgacctggtc cctgagtttc tgcctgcacc agaggatgtc 180

gaaacaaact gcgaatggag cgccttctcc tgttttcaga aggcccagct gaaatccgct 240

aacaccggca acaatgagcg aatcatcaac gtgagcatca agaagctgaa gcggaaaccc 300

cctagcacta atgctgggcg gagacagaaa catagactga cctgcccctc ttgtgacagt 360

tatgaaaaga aaccacccaa ggagttcctg gaacgcttta aaagtctgct gcagaaaatg 420

attcaccagc acctgtcctc cagaactcac gggtccgaag attcctaa 468

<210> 37

<211> 279

<212> DNA

<213> artificial sequence

<220>

<223> nucleic acid sequence of MIP-1alpha

<400> 37

atgcaggtgt caaccgccgc cctggctgtc ctgctgtgca ctatggctct gtgcaatcag 60

ttttccgcaa gtctggccgc tgatactccc accgcctgct gtttctctta cacaagtagg 120

cagatccctc agaacttcat tgctgactat tttgagacta gctcccagtg cagcaagccc 180

ggcgtgatct ttctgaccaa gcggagccgg caggtctgtg ccgatccctc cgaagaatgg 240

gtgcagaagt atgtctccga cctggaactg tcagcataa 279

Claims

1. A fusion protein comprising a fusion polypeptide derived from mycobacterium tuberculosis (Mycoba cterium tuberculosis).

2. The fusion protein of claim 1, wherein the fusion polypeptide is selected from the group consisting of Ag85A, TB10.4, mtb32b and PstS3.

3. The fusion protein of claim 2, wherein Ag85A consists of the amino acid sequence of seq id No. 1; the TB10.4 consists of the amino acid sequence of SEQ ID NO. 2; the Mtb32b consists of the amino acid sequence of SEQ ID NO. 3; the PstS3 consists of the amino acid sequence of SEQ ID NO. 4.

4. The fusion protein of claim 1, wherein the fusion polypeptide further comprises a polypeptide selected from the group consisting of PPE39, glcB and RipA.

5. The fusion protein of claim 4, wherein the PPE39 consists of the amino acid sequence of SEQ ID NO. 5; the above GlcB consists of the amino acid sequence of seq id No. 6; the RipA consists of the amino acid sequence of SEQ ID No. 7.

6. The fusion protein of claim 2 or 4, wherein the fusion polypeptide is linked via a GS linker.

7. The fusion protein of claim 6, wherein the GS linker consists of the amino acid sequence of seq id No. 8.

8. The fusion protein of claim 1, further comprising a signal peptide.

9. The fusion protein of claim 8, wherein the signal peptide is selected from the group consisting of a tissue-type plasminogen activator signal peptide, a herpes simplex virus glycoprotein Ds signal peptide, and a growth hormone signal peptide.

10. The fusion protein of claim 1, wherein the fusion protein further comprises an immunopotentiating peptide.

11. The fusion protein of claim 10, wherein the immunopotentiating peptide is selected from the group consisting of CD40 ligand, FMS-like tyrosine kinase 3 ligand, flagellin, and OX 40.

12. A polynucleotide for encoding the fusion protein according to any one of claims 1 to 11.

13. A recombinant vector comprising the polynucleotide of claim 12.

14. The recombinant vector according to claim 13, wherein the recombinant vector is a plasmid vector or a viral vector.

15. A pharmaceutical composition for preventing or treating tuberculosis, characterized by comprising the fusion protein according to any one of claims 1 to 11.

16. The composition of claim 15, wherein the composition further comprises an immunoadjuvant comprising interleukin-12, interleukin-21, and macrophage inflammatory protein-1 alpha.

17. The composition of claim 16, wherein said immunoadjuvant comprises the amino acid sequence of seq id No. 30 to 33.

18. A pharmaceutical composition for preventing or treating tuberculosis, comprising the polynucleotide of claim 12.

19. The composition of claim 18, wherein the composition further comprises a polynucleotide encoding an immunoadjuvant comprising interleukin-12, interleukin-21, and macrophage inflammatory protein-1 a.

20. A pharmaceutical composition for preventing or treating tuberculosis, comprising the recombinant vector of claim 13.