CN107096015A - The antivirus action of novel antiviral molecule Kdm6a a kind of and its application - Google Patents

Abstract

The present invention relates to biotechnology and medical domain, specifically a kind of novel antiviral molecule Kdm6a antivirus action and its application.Present invention relates particularly to the purposes containing Kdm6a albumen or Kdm6a coded sequences, its accelerator or its inhibitor in the medicine or kit of the generation of I types interferon is regulated and controled, and purposes and methods for using them and corresponding medicine, pharmaceutical composition or kit further to above-mentioned substance in prevention and treatment disease of viral infection.The present invention can be used for the generation of regulation and control I type interferon, relevant disease caused by resistance exogenous virus infection, and treatment virus infection, or for immunological regulation, be with a wide range of applications.

Description

The antivirus action of novel antiviral molecule Kdm6a a kind of and its application

Technical field

The present invention relates to biotechnology and medical domain, specifically, the present invention relates to the specific demethylation of lysine Enzyme 6a (Lysine-specific deMethylase 6a, Kdm6a) is led in resistance exogenous virus infection, and treatment virus infection Effect, mechanism of action, implementation and purposes in the relevant disease of cause.

Background technology

Virus is for core, using protein as shell by one kind with nucleic acid (RNA or DNA DNA) (capsid capsid) and composition molecule, its nucleic acid can be divided into sub-thread or bifilar, linear or spherical.Virus is minimum life Thing pathogen, from 20nm to 300nm：Maximum of which is acne viroid, and such as vaccinia virus is 300nm × 200nm, It is visible reluctantly under ordinary optical microscope；Medium sized such as influenza virus, a diameter of 80~100nm；Small-sized virus Such as epidemic encephalitis B virus, a diameter of 20nm, it is necessary to can just see under an electron microscope (basic virology, it is big gram strong Deng work, Chemical Industry Press, the first edition in 2005；Molecular Virology, yellow galaxy of literary talent etc. writes, People's Health Publisher, 2001 The first edition).

With virology and the progress of immunological investigation, understanding of the people to viral disease (Viral diseases) Gradually develop.The most frequently used virus taxis mode is that virus is classified according to the property (RNA or DNA) of nucleic acid, typically Virus is divided into DNA virus and RNA virus.Common pathogenic DNA virus mainly has human papilloma virus (HPV), hepatitis B Viral (HBV), Epstein-Barr virus etc., common pathogenic RNA virus include epidemic hemorrhagic fever virus, people's acquired immunodeficiency disease (basic virology, big gram strong wait writes, chemistry for malicious (HIV), serious acute respiratory syndrome (SARS) virus and influenza virus etc. Industrial publishing house, the first edition in 2005；Molecular Virology, yellow galaxy of literary talent etc. writes, People's Health Publisher, the first edition in 2001).Disease Viral disease can be divided into Respirovirus, arboviruse, eruptive virus, enteron aisle disease according to clinical manifestation and spread path Infection etc. caused by malicious.

Viral disease often causes than more serious clinical manifestation, often causes outbreak of epidemic.Virus infection after except Cause in a short time outside the local inflammation caused by infection, long-term virus infection can cause tumour and organ failure etc. Serious disease, the cervical carcinoma as caused by HPV infection, liver cancer, hepatic sclerosis and liver failure, Epstein-Barr virus caused by HBV infection Respiratory failure and stream caused by immunodeficiency caused by caused nasopharyngeal carcinoma, HIV and Kaposi's sarcoma, SARS (basic virology, big gram strong wait writes respiratory dysfunction caused by Influenza Virus etc., Chemical Industry Press, the first edition in 2005； Molecular Virology, yellow galaxy of literary talent etc. writes, People's Health Publisher, the first edition in 2001).

The infection of each epidemic virus can cause the loss of great health and national economy, but so far Specific drug is there is no to antiviral, and clinical treatment emphasis is just being suited the medicine to the illness and expectant treatment, therefore the molecule of the morbidity of viral disease The research of research and therapeutic strategy with amynologic mechanism has great scientific meaning and display meaning.

Interferon (interferon, IFN) is one group of trace regulation albumen existing in vivo, having extensive bioactivity, With broad-spectrum antiviral, suppressing cell reproduction and immunoregulatory activity, clinic is had been widely used for.Interferon is place after virus infection The glycoprotein with ntiviral characteristic of chief cell release.According to its biology, biochemistry, gene expression characteristicses and cell derived, Four types, i.e. α, β, γ, ω type interferon can be divided into, leucocyte (Monocytes/Macrophages) is respectively derived from, into fiber finer Born of the same parents, lymphocyte (T-cell and trophocyte).Interferon has 3 kinds of functions of antiviral, antiproliferative and immunological regulation.Wherein α, β type IFN are also known as I type interferon, have critical role in the treatment of viral disease.

Interferon has been widely used for the treatment of viral disease, such as herpes zoster, herpetic oropharynx, child virus Pneumonia, Japanese Type-B encephalitis, virus hepatitis, AIDS, condyloma acuminatum, SARS etc., can suppress duplication and the tune of virus The generation of virus specific immune response is saved, is to be able to prove and commercial applications having in clinical disease toxic treatment at present Imitate biological agent.

People recognize that the molecular mechanism of adventitious viruses not fully understands for body, but with the hair of Toll-like molecule Existing, body is progressively recognized for the generation mechanism and mechanism of action of the IFN produced by virus.Research finds that TLR3 is main Recognize viruses molecule double center chain RNA (double-stranded RNA, dsRNA) (Takeuchi, O. etc., Immunity.1999； 11:443-451；Ozinsky, A. etc., Proc Natl Acad Sci USA.2000；97:13766-13771； Alexopoulou, L. etc., Nature.2001；413:732-738)；The mankind TLR7 and the TLR8 assignment of genes gene mapping in chromosome x p22, Recognize single stranded RNA (single-stranded RNA, ssRNAs) (Heil, F. etc., Science.2004 in viruses molecule； 303:1526-1529)；Mankind TLR9 is positioned at chromosome 3p 21.3, identification bacterium and the CpG motifs or artificial in viral DNA The CpG of synthesis few core former times acid (CpG oligodeoxynucleotides, CpG ODN) (Hemmi, H. etc., Nature.2000； 408:740-745).In addition, virus can also be produced by TLR non-dependent modes I types interferon (Baccala, R. etc., Nat.Med.13,543-551(2007)).Wherein, TLR3 can be lured by TRIF-TBK1/IKK ε-IRF3 signal transduction pathway Lead the generation of I type interferon；TLR7/8/9 can activate the generation of inducing type I interferon by MyD88-IRF7 signal paths； In addition, some enter the TBK1/IKK ε-IRF3 signal paths activation that intracellular virus can be mediated by RIG-I/MDA-5 Pass through the generation (Baccala, R. etc., Nat.Med.13,543-551 (2007)) of TLR non-dependent mode inducing type I interferons. Can inducing type I interferon generation and can suppress virus duplication and amplification be examine medicine therapeutic virus disease Very effective standard.

Kdm6a also known as UTX, can the specific methyl group for removing H3K27me2/3, performance H3K27 demethylation enzyme activity Property (Lee, MG. etc., Science 318,447-450 (2007)).The molecule possesses highly conserved homologous C-terminal domain I.e. JMJC domains, are also its enzymatic activity domain, main to play histone demethylation activity.The N-terminal of the molecule is included The TPR domains of 6 repetitive sequences, the main function of playing protein protein interaction.The Kdm6a assignments of genes gene mapping are dyed in X Body, but do not influenceed by x chromosome inactivation, in cell and body wide expression (Agger, K. etc., Nature 449, 731-734(2007))。

Kdm6a specific can remove methylating for H3K27me2/3 as important apparent modification enzyme, in point of individual (Lan, F. etc., Nature 449,689- are played an important role in change, development and the generation of tumor tissues, evolution 694 (2007)), but the concern that the function in immune system is subject to is considerably less.

In summary, this area is effective against virus infection in the urgent need to developing one kind, suppresses inflammatory factor generation Immunologic competence material.

The content of the invention

It is an object of the invention to provide Kdm6a albumen, Kdm6a coded sequences, its accelerator or its inhibitor in regulation and control I The aborning purposes of type interferon, and further provide their purposes in prevention and treatment disease of viral infection.This Medicine, pharmaceutical composition or the kit of invention can be used for the generation for promoting I type interferon, be effective against exogenous virus infection, With relevant disease caused by treatment virus infection.

The first aspect of the present invention is there is provided Kdm6a albumen or Kdm6a coded sequences, its accelerator or its inhibitor in system It is ready for use on the purposes in the medicine of regulation and control I types interferon generation or kit.

It is preferred that, the Kdm6a albumen is selected from：

(a)SEQ ID NO：2 or SEQ ID NO：Amino acid sequence shown in 4；Or

(b) with SEQ ID NO：2 or SEQ ID NO：Amino acid sequence homologous shown in 4, it, which has, suppresses virus infection The albumen or polypeptide of relevant disease and/or symptom activity；Or

(c) pass through substitution in the amino acid sequence of (a) or (b), lack or add one or several amino acid and with pre- Anti- or treatment virus infection relevant disease and/or the protein as derived from (a) or (b) or polypeptide of symptom activity.

It is furthermore preferred that the Kdm6a albumen is selected from：SEQ ID NO：2 or SEQ ID NO：Amino acid sequence shown in 4.

It is preferred that, the Kdm6a encoding genes are selected from：

(i)SEQ ID NO：1 or SEQ ID NO：1 378-4583 bit sequences, SEQ ID NO：3 or SEQ ID NO： Nucleotide sequence shown in 3 432-4637 bit sequences；Or

(ii) molecule under strict conditions with (i) nucleotide sequence hybridization limited；Or

(iii) by replacing, lacking or add one or several nucleotides and compile in (i) or (ii) nucleotide sequence Code has the molecule of prevention or treatment virus infection relevant disease and/or symptom.

It is furthermore preferred that the Kdm6a encoding genes are selected from：SEQ ID NO：1 or SEQ ID NO：378-4583 of 1 Sequence, SEQ ID NO：3 or SEQ ID NO：Nucleotide sequence shown in 3 432-4637 bit sequences.

It is preferred that, the accelerator of Kdm6a or its coded sequence is selected from：The overexpression of Kdm6a or its coded sequence is carried Body, exogenous Kdm6a, Kdm6a or its coded sequence naked DNA, the liposome DNA of Kdm6a or its coded sequence, Kdm6a albumen.

It is preferred that, the inhibitor of Kdm6a or its coded sequence is selected from for Kdm6a or its coded sequence：Antibody, SiRNA, miRNA, ASON, antagonist, blocking agent.

It is preferred that, the Kdm6a albumen is：Native purified albumen, the product of chemical synthesis or using recombinant technique from Produced in protokaryon or eucaryon host, be preferably people or mouse Kdm6a, be more preferably people's Kdm6a albumen.

It is preferred that, the host is selected from：Bacterium, yeast, higher mammal, insect and mammalian cell.

It is preferred that, the Kdm6a, its coded sequence or its accelerator promote the generation of I type interferon；The inhibitor suppression The generation of I types interferon processed.The I types interferon is selected from：IFN-α or IFN-β, preferably IFN-β.

It is preferred that, the medicine or kit include Kdm6a or Kdm6a coded sequences or its accelerator, and by promoting The generation of I type interferon is further used for preventing or treats the disease related to viral infection and/or its symptom；Or

The inhibitor of the medicine or kit comprising Kdm6a or its coded sequence, and by suppressing the mistake of I type interferon Degree, which is produced, is used to prevent or treat tissue and organ damage and/or its symptom caused by I type interferon overexpressions.

It is preferred that, the virus infection relevant disease and/or symptom are the one or more being selected from the group because of virus Disease and/or symptom caused by infection：Virus is metainfective to be replicated；The local cell of virus infection is (epithelial cell of skin, outer The composition cell of the immunocyte of all blood, the immunocyte in immune organ and solid organ) and tissue (including skin, liver Dirty, kidney, brain, lungs) damage；Organ (including skin, liver, kidney, brain, lungs) function damage that virus infection is caused.

It is preferred that, the virus infection relevant disease and/or symptom are selected from：Herpes zoster, herpetic oropharynx, children's virus Property pneumonia, Japanese Type-B encephalitis, virus hepatitis, AIDS, condyloma acuminatum, SARS.

It is preferred that, the virus infection is caused by the one or more virus being selected from the group：People's papilloma Virus, hepatitis B, Epstein-Barr virus, epidemic hemorrhagic fever virus, people's acquired immunity defact virus, serious acute respiratory syndrome Virus, influenza virus.

It is preferred that, the generation of the Kdm6a albumen and its coded sequence inducing type I interferon, so as to prevent or treat disease Poison infection relevant disease and/or symptom.

It is preferred that, the medication of the medicine is selected from：Give Kdm6a coded sequences, such as direct naked DNA injection method, Liposome DNA direct injections, gold coating DNA particle bombardments, breeding unsoundness bacterium carry DNA method, replicated Defect adenovirus carries target DNA method；Give Kdm6a albumen, for example drug administration by injection (such as direct injection Kdm6a albumen or uses fat Plastid embedding Kdm6a albumen), nasal-cavity administration, pulmonary administration, oral administration, cutaneous penetration (such as electro-ionic osmosis).

The second aspect of the present invention is used to regulate and control the pharmaceutical composition that I types interferon is produced there is provided a kind of, and it is included：

(A) the Kdm6a albumen or Kdm6a coded sequences of therapeutically effective amount, its accelerator or its inhibitor；

(B) prevent or treat virus infection relevant disease and/or other active materials of symptom activity；And

(C) acceptable carrier or excipient pharmaceutically or in immunology.

It is preferred that, Kdm6a albumen, its coded sequence, its accelerator or its inhibitor account for medicine in described pharmaceutical composition 0.001~99.9wt% of composition total weight.

It is preferred that, Kdm6a albumen, its coded sequence, its accelerator or its inhibitor account for medicine in described pharmaceutical composition 1~95wt% of composition total weight, preferably 5~90wt%, more preferably 10~80wt%.

It is preferred that, prior to, concurrently with, or after the pharmaceutical composition of the present invention is given, give with prevention or treatment virus Infect other active materials of relevant disease and/or symptom activity.It is described have prevention or treatment virus infection relevant disease and/ Or other active materials of symptom activity are selected from：The conventional antiviral drugs of clinic (including anti-influenza A virus surface M2 acceptors, It is anti-neuraminidase, anti-monophosphate carnine acidohydrogenase, antiviral DNA polymerases, AntiHIV1 RT activity reverse transcriptase, anti- Hiv protease, anti-sialidase, anti-unwindase) one or more.

There is provided a kind of prevention or the side for the treatment of virus infection relevant disease and/or its symptom for the third aspect of the present invention Method, methods described includes：Give the Kdm6a albumen of object effective dose for needing to prevent or treat, its coded sequence, and/or its Accelerator or the above-mentioned pharmaceutical composition of effective dose.

It is preferred that, the virus infection relevant disease and/or symptom are the one or more being selected from the group because of virus infection Caused disease and/or symptom：Virus is metainfective to be replicated；Local cell (epithelial cell, the peripheral blood of skin of virus infection Immunocyte, the composition cell of the immunocyte in immune organ and solid organ) and tissue (including skin, liver, kidney Dirty, brain, lungs) damage；Organ (including skin, liver, kidney, brain, lungs) function damage that virus infection is caused.

It is preferred that, the virus infection is caused by the one or more virus being selected from the group：Human papilloma virus, Hepatitis B, Epstein-Barr virus, epidemic hemorrhagic fever virus, people's acquired immunity defact virus, SARS virus, Influenza virus.

It is preferred that, the I types interferon is selected from：IFN-α or IFN-β.

The fourth aspect of the present invention there is provided one kind prevention or treat caused by I type interferon overexpressions tissue and The method of organ damage and/or its symptom, methods described includes：Give the Kdm6a for the object effective dose for needing to prevent or treating The inhibitor of albumen or its coded sequence, or effective dose above-mentioned pharmaceutical composition.

The invention has the advantages that：

1st, present invention is disclosed Kdm6a and its New function of coded sequence, that is, I types IFN generation is effectively facilitated, resistance is outer The function of relevant disease caused by source virus infection, and treatment virus infection；

2nd, the invention provides a kind of generation for effectively facilitating I types IFN, resistance exogenous virus infection, and treatment virus The medicine of relevant disease caused by infection.

Brief description of the drawings

Fig. 1：The promotion effect that the IFN-β that Kdm6a eukaryotic expression vector transfections Turnover of Mouse Peritoneal Macrophages is induced VSV is produced Really.Wherein, Figure 1A is quantitative RT PCR analysis result；Figure 1B is elisa assay result.

Fig. 2：The inhibition that interference Kdm6a is produced in the IFN-β that the expression of Turnover of Mouse Peritoneal Macrophages is induced VSV. Wherein, Fig. 2A is quantitative RT PCR analysis；Fig. 2 B are elisa assay." siCtrl " represents out of order control RNA interfering (scrambled control RNA interference)；" siKdm6a " represents the specific RNA interferings of Kdm6a.

Embodiment

The embodiment provided with reference to embodiment the present invention elaborates.It should be understood that these embodiments It is only illustrative of the invention and is not intended to limit the scope of the invention.Those skilled in the art can make appropriate repair to the present invention Change, change, these modifications and variation are within the scope of the present invention.

The experimental method of unreceipted actual conditions in the following example, can use the conventional method in this area, for example, join Examine Sambrook et al.《Molecular Cloning:A Laboratory guide》(New York：Cold Spring Harbor Laboratory Press, 1989), or according to the condition proposed by manufacturer.DNA sequence measurement is the conventional method in this area, also can be by Commercial company provides test.

Unless otherwise indicated, otherwise percentage and number are calculated by weight.

Unless otherwise defined, all specialties used in text known to scientific words and one skilled in the art with anticipating Justice is identical.In addition, any method similar or impartial to described content and material all can be applied in the present invention.Described in text Preferable implementation only present a demonstration and be used with material.

The present inventor has found Kdm6a in disease of viral infection by substantial amounts of research, with promotion I type interferon Generation function.The present inventor completes the present invention on this basis.

Specifically, it is viral molecular biology and cell biology to carry out application study for IFN regulation related genes The IFN prevention and treatment for promoting the nucleotides and protein of gene to be applied to viral disease are artificial dry by the focus of research The effective technology of pre- viral infection, therefore either in terms of functional genome research, or viral relatively gene therapy It is respectively provided with extensively application prospect.

Inventor has found that generation and the function and its TPR of I type interferon can be promoted by being overexpressed Kdm6a by studying The interactional function of domain is closely related.Interference Kdm6a expression can suppress the generation of I type interferon.Point out Kdm6a May possess the application prospect of viral disease.Therefore the invention also discloses apply the antiviral molecule in viral infection disease The method and strategy of the prevention and treatment of disease.

All number ranges provided herein be intended to clearly include falling all numerical value between endpoints of ranges and it Between number range.The feature that the feature or embodiment that can be mentioned to the present invention are mentioned is combined.This specification is taken off All features shown can be used in combination with any combinations thing form, and each feature disclosed in specification can provide phase with any The alternative characteristics substitution of same, impartial or similar purpose.Therefore except there is special instruction, disclosed feature is only impartial or similar The general example of feature.

As used herein, " containing ", " having " or " comprising " include "comprising", " mainly by ... constitute ", " substantially By ... constitute " and " by ... constitute "；" mainly by ... constitute ", " substantially by ... constitute " and " by ... constitute " Belong to the subordinate concept of " containing ", " having " or " comprising ".

Kdm6a albumen (polypeptide)

As used herein, term " Kdm6a albumen (polypeptide) ", " Kdm6a albumen (polypeptide) ", " Kdm6a ", " UTX " can be mutual Change and use, a kind of its histone H 3 K27me2/3 demethylase.The Kdm6a albumen of the present invention can be by SEQ ID NO：1 Sequence (people's full length sequence) or SEQ ID NO:1 378-4583 bit sequences (people CDS sequences) or SEQ ID NO:3 (mouse Full length sequence) or SEQ ID NO:Protein or these albumen coded by 3 432-4637 bit sequences (mouse CDS sequences) The homologous sequence that matter has antivirus action (for example can obtain Kdm6a's by database known in the art or comparison software Homologous sequence), variant or modified forms.For example, the Kdm6a albumen may be selected from：(a)SEQ ID NO：2 or SEQ ID NO:Amino acid sequence shown in 4；Or (b) passes through substitution, missing or addition one or several in the amino acid sequence that (a) is limited Individual amino acid and with suppress inflammatory factor it is active as protein derived from (a) or polypeptide.

The protein or polypeptide of the present invention can be native purified product, or chemical synthesis product, or use weight Group technology is produced from protokaryon or eucaryon host (for example, bacterium, yeast, higher mammal, insect and mammalian cell).This hair Bright middle Kdm6a albumen or polypeptide are preferably encoded by people Kdm6a genes or its homologous gene or family gene.

The variant form of present protein or polypeptide includes (but being not limited to)：One or more (it is usually 1-50, Preferably 1-30, more preferably 1-20, most preferably 1-10, such as 1,2,3,4,5,6,7,8,9 or 10) amino acid lacks Lose, insert and/or replace, and it (is usually within 20, preferably to add one or several in C-terminal and/or N-terminal Within 10, more preferably within 5) amino acid.For example, in the art, being carried out with similar nature or similar amino acid During substitution, it will not generally change the function of protein or polypeptide.Again such as, one or several is added in C-terminal and/or N-terminal Amino acid will not generally also change the function of protein or polypeptide, Kdm6a protein or polypeptide of the invention may include or Do not include the methionine residues of starting and still there is antiviral activity.

It using radiation or can get off to produce random mutagenesis exposed to mutagens, can also pass through site-directed mutagenesis or other known Protocols in Molecular Biology obtain the protein or polypeptide in above-mentioned (b).Using code for said proteins or the volume of polypeptide Code sequence builds transgenic animals, and observe the transgenic animals whether the resistance of virus infection has been improved screening and Differentiate gained protein or polypeptide.

Host according to used in recombinant production scheme, protein of the invention or polypeptide can be glycosylated, or can be with It is nonglycosylated.The term also includes the active fragment and reactive derivative of Kdm6a albumen.

The variant form of the polypeptide includes：Homologous sequence, conservative variant, allelic variant, natural mutation, induction Albumen coded by mutant, the sequence that can hybridize with Kdm6a albumen coded sequences under the conditions of the high or low stringency and The many peptide or proteins obtained using the antiserum of anti-Kdm6a albumen.The present invention it is also possible to use other polypeptides, such as comprising Kdm6a eggs The fusion protein of white or its fragment.In addition to the almost polypeptide of total length, present invention includes the soluble piece of Kdm6a albumen Section.Generally, the fragment has at least about 10 continuous amino acids of Kdm6a protein sequences, typically at least about 30 continuous amino Acid, preferably at least about 50 continuous amino acids, more preferably at least about 80 continuous amino acids, most preferably at least about 100 companies Continuous amino acid.

Kdm6a protein coding genes

As used herein, term " Kdm6a genes " or " Kdm6a albumen coded sequences " are used interchangeably, and each mean one kind The sequence of Kdm6a albumen of the present invention or polypeptide is encoded, it can be SEQ ID NO：1 (people's total length) or SEQ ID NO：1 378-4583 (people CDS) sequences, SEQ ID NO：3 (mouse total lengths) or SEQ ID NO：432-4637 of 3 are (small Mouse CDS) nucleotide sequence shown in sequence, under strict conditions with the molecules of these sequence hybridizations or with above-mentioned numberator height Homologous family gene molecule, the expression of the gene has certain inhibitory action to virus infection.The Kdm6a bases of the present invention Because may be selected from：(i)SEQ ID NO：1 or SEQ ID NO：1 378-4583 bit sequences, SEQ ID NO：3 or SEQ ID NO：Nucleotide sequence shown in 3 432-4637 bit sequences；Or the sequence hybridization that (ii) is limited with (i) under strict conditions And with the molecule for suppressing inflammatory factor activity.

As used herein, term " stringent condition " refers to：(1) hybridization under compared with low ionic strength and higher temperature and wash It is de-, such as 0.2 × SSC, 0.1%SDS, 60 DEG C；Or added with denaturant during (2) hybridization, such as 50% (v/v) formamide, 0.1% calf Serum/0.1%Ficoll, 42 DEG C etc.；Or the phase same sex of (3) only between two sequences at least 50%, preferably more than 55%, More than 60%, more than 65%, more than 70%, more than 75%, more than 80%, more than 85% or more than 90%, more preferably 95% Just hybridize during the above.For example, the sequence can be the complementary series of sequence defined in (a).

The Kdm6a gene nucleotides full length sequence or its fragment of the present invention can generally use PCR TRAPs, recombination method or people The method of work synthesis is obtained.For PCR TRAPs, can especially it be opened according to relevant nucleotide sequence disclosed in this invention Reading frame sequence designs primer, and with commercially available cDNA storehouses or as prepared by conventional method well known by persons skilled in the art CDNA storehouses obtain relevant sequence as template, amplification.When sequence is longer, it is often necessary to carry out twice or multiple PCR is expanded, so The fragment for afterwards again amplifying each time is stitched together by proper order.

It should be understood that the Kdm6a genes of the present invention are preferably obtained from people, it is highly same obtained from other animals and people Kdm6a genes Source (as having more than 50%, preferably more than 55%, more than 60%, more than 65%, more than 70%, more than 75%, more than 80%, More preferably more than 85% such as 85%, 90%, 95%, even 98% sequence thereto) other genes also preferably examined in the present invention Within the equivalency range of worry.The Method and kit for of the aligned sequences phase same sex is also well known in the art, such as BLAST.

The accelerator and inhibitor of Kdm6a or its coded sequence

Kdm6a or " accelerator " of its coded sequence are further related in the present invention.Term " accelerator " or " Kdm6a or its volume The accelerator of code sequence " is used interchangeably, and refers to the material that can improve Kdm6a or the level or activity of its coded sequence.It can use Accelerator in the present invention includes but is not limited to：Kdm6a expression vectors, exogenous Kdm6a, Kdm6a or its coded sequence Liposome DNA, the Kdm6a albumen of naked DNA, Kdm6a or its coded sequence.

The Kdm6a of the present invention, its coded sequence or its accelerator promote the generation of I type interferon be further used for prevention or The treatment disease related to viral infection and/or its symptom；

Kdm6a or " inhibitor " of its coded sequence are further related in the present invention.Term " inhibitor " or " Kdm6a or its volume The inhibitor of code sequence " is used interchangeably, and refers to the material that can reduce Kdm6a or the level or activity of its coded sequence.It can use Inhibitor in the present invention includes but is not limited to, for Kdm6a or its coded sequence：Antibody, siRNA, miRNA, antisense Oligonucleotides, antagonist, blocking agent.

The excessive generation that the inhibitor of the present invention can suppress I type interferon is used to prevent or treat because I types interferon is excessive Tissue and organ damage and/or its symptom caused by expression.

Carrier, host and transgenic animals

The invention further relates to include the carrier of Kdm6a genes, and the host cell produced with the carrier through genetic engineering, And high expressing K dm6a transgenic animals are obtained by transgenosis.

Pass through conventional recombinant DNA technology (Science, 1984；224：1431), can using the coded sequence of the present invention For expressing or producing the Kdm6a albumen of restructuring.In general there are following steps：

(1) with the polynucleotides (or variant) of the encoded K dm6a albumen of the present invention, or with the weight containing the polynucleotides The conversion of group expression vector or suitable host cell of transduceing；

(2) host cell cultivated in suitable culture medium；With

(3) separation, protein purification or polypeptide from culture medium or cell.

In the present invention, term " carrier " is used interchangeably with " recombinant expression carrier ", refer to bacterial plasmid well known in the art, Bacteriophage, yeast plasmid, zooblast virus, mammalian cell virus or other carriers.In a word, as long as can be in host Replicate and stably, any plasmid and carrier can be used.One key character of expression vector is to usually contain replication orgin, open Mover, marker gene and translation control element.

Method well-known to those having ordinary skill in the art can be used to build coded sequence containing Kdm6a and suitable transcription/translation The expression vector of control signal.These methods include recombinant DNA technology in vi, DNA synthetic technologys, In vivo recombination technology etc..Institute The DNA sequence dna stated can be effectively connected in the appropriate promoter in expression vector, to instruct mRNA to synthesize.Expression vector also includes The ribosome bind site and transcription terminator of translation initiation.PcDNA3.1 carriers, pIRES2- are preferably used in the present invention EGFP carriers, Adeno-X expression systems and pSilencer 2.1-U6-neo interference carriers.

In addition, expression vector preferably includes one or more selected markers, it is used to select conversion to provide Dihyrofolate reductase, neomycin resistance and the green fluorescence egg of the phenotypic character of host cell, such as eukaryotic culture In vain (GFP), or tetracycline or amicillin resistance for Escherichia coli.

The carrier of above-mentioned appropriate DNA sequence dna and appropriate promoter or control sequence is included, can be used for conversion suitable When host cell, allow it to marking protein or polypeptide.Host cell can be prokaryotic, such as bacterial cell；Or It is low eukaryotic, such as yeast cells；Or higher eucaryotic cells, such as zooblast.Representative example has：Escherichia coli, Streptomyces, Agrobacterium；Fungal cell's such as yeast；Zooblast etc..In the present invention, it is preferred to thin using E. coli bacteria Born of the same parents, mouse macrophage are used as host cell.

When the polynucleotides of the present invention are expressed in higher eucaryotic cells, if will when inserting enhancer sequence in the carrier Transcription can be strengthened.Enhancer is DNA cis-acting factors, generally about has 10 to 300 base-pairs, acts on and open Mover is to strengthen the transcription of gene.Persons skilled in the art are aware that how to select appropriate carrier, promoter, enhancer And host cell.

Term " transgenic animals " or " transformed animal " are used interchangeably in the present invention, are referred both to by conventional transgenic The cell, organ, the tissue or individual that are transferred to Kdm6a genes of the present invention and stablize high expressing K dm6a albumen or polypeptide that method is obtained Body.

Recombinant polypeptide in the above methods can express or be secreted into the cell or on cell membrane extracellular.If Need, can be separated using its physics, chemistry and other characteristics by various separation methods and purification of Recombinant albumen.These Method is well-known to those skilled in the art.The example of these methods includes but is not limited to：Conventional renaturation process, use egg White precipitating reagent processing (salting-out method), centrifugation, the broken bacterium of infiltration, super processing, ultracentrifugation, sieve chromatography (gel filtration), absorption Chromatography, ion-exchange chromatography, the combination of high performance liquid chroma- tography (HPLC) and other various liquid chromatography technologies and these methods.

Medicine, pharmaceutical composition or kit

Present invention also offers a kind of medicine, pharmaceutical composition or kit, wherein containing the of the invention of effective dose Kdm6a or Kdm6a coded sequences, its accelerator or inhibitor, and acceptable carrier pharmaceutically or in immunology.As herein Used, term " active material " or " active material of the invention " are used interchangeably, refer to Kdm6a or Kdm6a coded sequences, its Accelerator or inhibitor.

In preferably embodiment, described pharmaceutical composition can be used for treatment to be treated or in advance with known in the state of the art Preventing virus infection relevant disease and/or symptom, for example：Virus is metainfective to be replicated；Virus is metainfective to be replicated；Viral infection office Cell (epithelial cell of skin, the immunocyte of peripheral blood, the group of the immunocyte in immune organ and solid organ in portion Into cell) and tissue (including skin, liver, kidney, brain, lungs) damage；Organ (including skin, liver that virus infection is caused Dirty, kidney, brain, lung, spleen) function damage.

In the present invention, the virus infection relevant disease and/or symptom may be selected from：It is herpes zoster, herpetic oropharynx, small Youngster's viral pneumonia, Japanese Type-B encephalitis, virus hepatitis, AIDS, condyloma acuminatum, SARS.

Virus infection can be caused by the one or more virus being selected from the group：Human papilloma virus, hepatitis B, EB Virus, epidemic hemorrhagic fever virus, people's acquired immunity defact virus, SARS virus, influenza virus.

As used herein, term " containing " or " comprising " include "comprising", " substantially by ... constitute " and " by ... constitute ".

As used herein, term " pharmaceutically acceptable " composition apply to people and/or animal and without excessive bad pair Reaction (such as toxicity, stimulation and allergy), that is, have rational benefit/risk than material.

As used herein, term " effective dose " refer to that people and/or animal can be produced function or activity and can by people and/ Or the amount that animal is received.

As used herein, term " pharmaceutically acceptable carrier " refers to the carrier for Therapeutic Administration, including various taxes Shape agent and diluent.The term refers to some such medicament carriers：Themselves it is not necessary active component, and does not have after administration There is undue toxicity.Suitable carrier is well known to those of ordinary skill in the art.《Remington pharmaceutical science》 It can be found on pharmaceutically in (Remington ' s Pharmaceutical Sciences, Mack Pub.Co., N.J.1991) Acceptable excipient is discussed fully.

Pharmaceutically acceptable carrier can contain liquid, such as water, salt solution, glycerine and ethanol in the composition.In addition, these Complementary material, such as filler, disintegrant, lubricant, glidant, effervescent agent, wetting agent or breast are there is likely to be in carrier Agent, flavouring, pH buffer substance etc..Generally, these materials can be formulated in nontoxic, inert and pharmaceutically acceptable In aqueous carrier medium, wherein pH ordinarily be about 5-8, it is preferred that pH is about 6-8.

In the composition of the present invention Kdm6a albumen or its coded sequence active ingredient account for composition total weight 0.001~ 99.9wt%；Preferably 1~95wt% of composition total weight, is more preferably 5~90wt%, more preferably 10~80wt%.It is remaining Measure as the material such as pharmaceutically acceptable carrier and other additives.

As used herein, term " unit dosage forms " refers to, in order to convenient to take, the composition of the present invention is prepared into single Take required formulation, including but not limited to various solid formulations (such as tablet), liquid agent, capsule, sustained release agent.

In another preferred embodiment of the present invention, the composition is unit formulation or multi-form, and wherein Kdm6a The content of albumen or its coded sequence is 0.01~2000mg/ agent, preferably 0.1~1500mg/ agent, more preferably 1~1000mg/ Agent.In another preference of the present invention, daily using 1~6 dose of composition of the invention, preferably using 1~3 dose；It is optimal Choosing, the dosage taken daily is 1 dose.

It should be understood that the effective dose of Kdm6a albumen used or its coded sequence can be with the tight of to be administered or treatment object Weight degree and change.Concrete condition (such as object body weight, age, health, required reaches according to the individual instances of object Effect) determine, this is in the range of skilled practitioners may determine that.

The composition of the present invention, can for solid-state (such as granule, tablet, freeze-dried powder, suppository, capsule, sublingual lozenge) or Liquid (such as oral liquid) or other suitable shapes.Method of administration can be used：(1) direct naked DNA or protein injection method； (2) Kdm6a cDNA, mRNA and protein are connected with transferrins/poly-l-lysine compound, imitated with strengthening its biology Should；(3) cDNA, mRNA and protein and positively charged lipid formation compound, with caused by overcoming phosphate backbones negative electrical charge Pass through the difficulty of cell membrane；(4) enter cell with mediation after liposome cDNA, mRNA and protein, not only improve big point The smooth hydrolysis entered again from extracellular various enzymes of son；(5) cDNA, mRNA and protein are combined with cholesterol makes it The endochylema retention time increases by 10 times；(6) its unitransport can be made to target by transporting cDNA, mRNA and protein with immunoliposome Tissue and target cell；(7) also can be preferably to reprinting cell (such as fibroblast) by cDNA, mRNA and protein in-vitro transfection Kdm6a related drugs are loaded into target cell；(8) electricity punching (electroporation), i.e., by means of electric current by cDNA, MRNA and protein import target cell.

In addition, can also contain in the composition of the present invention is used to improving and treating other active matters of disease of viral infection Matter, described other active materials are selected from the group：The conventional antiviral drugs of clinic (including anti-influenza A virus surface M2 by Body, anti-neuraminidase, anti-monophosphate carnine acidohydrogenase, antiviral DNA polymerases, AntiHIV1 RT activity reverse transcriptase, AntiHIV1 RT activity protease, anti-sialidase, anti-unwindase) one or more.

The nucleotides related Kdm6a of the present invention and pharmaceutical grade protein each other can with use in conjunction, can also with it is other Medicine and treatment means joint, the prevention and treatment for disease of viral infection.

Embodiment 1：The facilitation that Kdm6a is produced to macrophage IFN-β

The dehydration thiol-acetic acid culture medium 2ml of intraperitoneal injection 3%, after 4 days, cervical dislocation puts to death mouse, 75% alcohol Immersion 5-10 minutes, mouse web portion upward, cuts off an osculum in ventrimeson, tears skin, peritonaeum is completely exposed.Then 20ml is used The culture medium of syringe injection 10ml serum-frees 1640, needle point under peritonaeum to abdominal cavity then side, and needle point is upward, avoids intestines and fat Fat, slow suction, now, peritoneal lavage fluid contains a large amount of macrophages.The peritoneal fluid of collection is put in 50ml centrifuge tube. Repeat this step once, common 20ml.Cell suspension 800g is centrifuged 5 minutes, abandons supernatant, is resuspended with the DMEM containing 10%FCS.Carefully Born of the same parents' suspension inserts culture plate, and 37 DEG C of cultures change liquid after 1 hour, and attached cell is the Turnover of Mouse Peritoneal Macrophages of fresh separated.

Using pcDNA3.1 (being purchased from Invitrogen companies) as carrier for expression of eukaryon, XhoI/BamHI digestions will be carried The Kdm6a in site SEQ ID NO:3 cDNA products insert the relevant position of the carrier, expand in E.Coli bacterial strain DH-5 α Increase the carrier, purifying obtains Kdm6a carrier for expression of eukaryon after sequencing identification.Transiently transfect (transfection reagent JetPei- Macrophage is purchased from Polyplus companies) mouse macrophage.After 48 hours, 0.1MOI work VSV viruses are added (Indiana Strain) (is purchased from ATCC, can also prepared using following literature methods：Li X,Zhang Q,Ding Y,Liu Y,Zhao D,Zhao K,Shen Q,Liu X,Zhu X,Li N,Cheng Z,Fan G,Wang Q,Cao X.Methyltransferase Dnmt3a upregulates HDAC9 to deacetylate TBK1 for activation of antiviral innate immunity.Nature Immunology.2016；17(7):806- 815.) after processing different time, cell and culture supernatant are collected, preparing cDNA is used for quantitative RT-PCR (94 DEG C, 30Sec；58 DEG C, 30Sec；72 DEG C, 30Sec；Totally 30 circulation) detection IFN-β mRNA level in-site；Or it is (used by ELISA with culture supernatant Kit is purchased from PBL companies) detection IFN-β protein level.

The result of IFN-β mRNA quantitative RT PCR analysis and the elisa assay of IFN-β is respectively such as Figure 1A and Figure 1B institutes Show.As a result show：Kdm6a carrier for expression of eukaryon can promote the generation of mouse macrophage IFN-β.

Embodiment 2：The inhibitory action for disturbing Kdm6a expression to produce macrophage IFN-β

With the RNA interfering (Si-Kdm6a) for Kdm6a and analogies control (Si- analogies) transfection (transfection reagent INTERFERin is purchased from Polyplus companies) mouse macrophage as obtained above, fresh RPMI 1640 is changed after 6 hours and is trained Support base.

RNA interfering (Si-Kdm6a) and analogies control (Si- analogies) for Kdm6a is purchased from Genephama companies, Si-Kdm6a sequence such as SEQ ID NO：5 and SEQ ID NO：Shown in 6, the sequence such as SEQ ID NO of Si- analogies：7 Hes SEQ ID NO：Increasing by 2 dT in 3' shown in 8, during synthesis makes sequence more stablize.

Particular sequence is as follows：

Si-Kdm6a sequences：

5'-GCUGCUACGAAUCUCUAAUTT-3'(Shunyi)；

5'-AUUAGAGAUUCGUAGCAGCTT-3'(antisenses).

Si- analogies sequences：

5'-UUCUCCGAACGUGUCACGUTT-3'(Shunyi)；

5'-ACGUGACACGUUCGGAGAATT-3 ' (antisense).

After 48 hours, the work VSV viruses for adding 0.1MOI (are purchased from ATCC, can also prepared using following literature methods：Li X,Zhang Q,Ding Y,Liu Y,Zhao D,Zhao K,Shen Q,Liu X,Zhu X,Li N,Cheng Z,Fan G, Wang Q,Cao X.Methyltransferase Dnmt3a upregulates HDAC9 to deacetylate TBK1 for activation of antiviral innate immunity.Nature Immunology.2016；17(7):806- 815.) after processing different time, cell and culture supernatant are collected, preparing cDNA is used for quantitative RT-PCR (94 DEG C, 30Sec；58 DEG C, 30Sec；72 DEG C, 30Sec；Totally 30 circulation) detection Kdm6a mRNA level in-sites, IFN-β mRNA level in-site；Or use culture supernatant Pass through ELISA (used kit is purchased from PBL companies) detection IFN-β protein level.

The result difference of Kdm6a mRNA level in-sites, IFN-β mRNA quantitative RT PCR analysis and the elisa assay of IFN-β As shown in Figure 2 A and 2 B.As a result show：Interference Kdm6a has produced inhibitory action in the expression of mouse macrophage to IFN-β.

The preferred embodiment to the invention is illustrated above, but the invention be not limited to it is described Embodiment, those skilled in the art can also make a variety of equivalent on the premise of without prejudice to the invention spirit Modification or replacement, these equivalent modifications or replacement are all contained in the application claim limited range.

SEQUENCE LISTING

<110>Second Military Medical University, PLA

<120>The antivirus action of novel antiviral molecule Kdm6a a kind of and its application

<130> /

<160> 8

<170> PatentIn version 3.3

<210> 1

<211> 5785

<212> DNA

<213>Homo sapiens (Homo sapiens)

<220>

<221> CDS

<222> (378)..(4583)

<400> 1

gtgtgacaca attacaacaa ctttgtgctg gtgccgggga agtttgtgtc tccaacgaat 60

cccctcagtg ctccccagcc ccgcgcgctc cggccgttcc cgccgtcccc gcctgtggct 120

gccccctgcc caaccccgcg atgtgaccct acagccgaaa gccgccgctg ccgacccggg 180

ggctccgcag cccctgccgc cgccgccgcc gccttcaccg ccgccgcgtt gggatttttc 240

gtcgccgccg cccgcggcgg aggaggaggc ggcgataaag ttggtgtgct ggtcccgcgc 300

gcagattggg ggcgtcactg cgggccccgg tccgaggggg ggtgtcggcg ttggagttgt 360

gaattcgctg cgtttcc atg aaa tcc tgc gga gtg tcg ctc gct acc gcc 410

Met Lys Ser Cys Gly Val Ser Leu Ala Thr Ala

1 5 10

gcc gct gcc gcc gcc gct ttc ggt gat gag gaa aag aaa atg gcg gcg 458

Ala Ala Ala Ala Ala Ala Phe Gly Asp Glu Glu Lys Lys Met Ala Ala

15 20 25

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Gly Lys Ala Ser Gly Glu Ser Glu Glu Ala Ser Pro Ser Leu Thr Ala

30 35 40

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Glu Glu Arg Glu Ala Leu Gly Gly Leu Asp Ser Arg Leu Phe Gly Phe

45 50 55

gtg aga ttt cat gaa gat ggc gcc agg acg aag gcc cta ctg ggc aag 602

Val Arg Phe His Glu Asp Gly Ala Arg Thr Lys Ala Leu Leu Gly Lys

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Ala Val Arg Cys Tyr Glu Ser Leu Ile Leu Lys Ala Glu Gly Lys Val

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gag tct gat ttc ttt tgt caa tta ggt cac ttc aac ctc tta ttg gaa 698

Glu Ser Asp Phe Phe Cys Gln Leu Gly His Phe Asn Leu Leu Leu Glu

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gat tat cca aaa gca tta tct gca tac cag agg tac tac agt tta cag 746

Asp Tyr Pro Lys Ala Leu Ser Ala Tyr Gln Arg Tyr Tyr Ser Leu Gln

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tct gac tac tgg aag aat gct gcc ttt tta tat ggt ctt ggt ttg gtc 794

Ser Asp Tyr Trp Lys Asn Ala Ala Phe Leu Tyr Gly Leu Gly Leu Val

125 130 135

tac ttc cat tat aat gca ttt cag tgg gca att aaa gca ttt cag gag 842

Tyr Phe His Tyr Asn Ala Phe Gln Trp Ala Ile Lys Ala Phe Gln Glu

140 145 150 155

gtg ctt tat gtt gat ccc agc ttt tgt cga gcc aag gaa att cat tta 890

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

160 165 170

cga ctt ggg ctt atg ttc aaa gtg aac aca gac tat gag tct agt tta 938

Arg Leu Gly Leu Met Phe Lys Val Asn Thr Asp Tyr Glu Ser Ser Leu

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aag cat ttt cag tta gct ttg gtt gac tgt aat ccc tgc act ttg tcc 986

Lys His Phe Gln Leu Ala Leu Val Asp Cys Asn Pro Cys Thr Leu Ser

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aat gct gaa att caa ttt cac att gcc cac tta tat gaa acc cag agg 1034

Asn Ala Glu Ile Gln Phe His Ile Ala His Leu Tyr Glu Thr Gln Arg

205 210 215

aaa tat cat tct gca aaa gaa gct tat gaa caa ctt ttg cag aca gag 1082

Lys Tyr His Ser Ala Lys Glu Ala Tyr Glu Gln Leu Leu Gln Thr Glu

220 225 230 235

aat ctt tct gca caa gta aaa gca act gtc tta caa cag tta ggt tgg 1130

Asn Leu Ser Ala Gln Val Lys Ala Thr Val Leu Gln Gln Leu Gly Trp

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atg cat cac act gta gat ctc ctg gga gat aaa gcc acc aag gaa agc 1178

Met His His Thr Val Asp Leu Leu Gly Asp Lys Ala Thr Lys Glu Ser

255 260 265

tat gct att cag tat ctc caa aag tcc ttg gaa gca gat cct aat tct 1226

Tyr Ala Ile Gln Tyr Leu Gln Lys Ser Leu Glu Ala Asp Pro Asn Ser

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ggc cag tcc tgg tat ttc ctc gga agg tgc tat tca agt att ggg aaa 1274

Gly Gln Ser Trp Tyr Phe Leu Gly Arg Cys Tyr Ser Ser Ile Gly Lys

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gtt cag gat gcc ttt ata tct tac agg cag tct att gat aaa tca gaa 1322

Val Gln Asp Ala Phe Ile Ser Tyr Arg Gln Ser Ile Asp Lys Ser Glu

300 305 310 315

gca agt gca gat aca tgg tgt tca ata ggt gtg cta tat cag cag caa 1370

Ala Ser Ala Asp Thr Trp Cys Ser Ile Gly Val Leu Tyr Gln Gln Gln

320 325 330

aat cag ccc atg gat gct tta cag gcc tat att tgt gct gta caa ttg 1418

Asn Gln Pro Met Asp Ala Leu Gln Ala Tyr Ile Cys Ala Val Gln Leu

335 340 345

gac cat ggc cat gct gca gcc tgg atg gac cta ggc act ctc tat gaa 1466

Asp His Gly His Ala Ala Ala Trp Met Asp Leu Gly Thr Leu Tyr Glu

350 355 360

tcc tgc aac cag cct cag gat gcc att aaa tgc tac tta aat gca act 1514

Ser Cys Asn Gln Pro Gln Asp Ala Ile Lys Cys Tyr Leu Asn Ala Thr

365 370 375

aga agc aaa agt tgt agt aat acc tct gca ctt gca gca cga att aag 1562

Arg Ser Lys Ser Cys Ser Asn Thr Ser Ala Leu Ala Ala Arg Ile Lys

380 385 390 395

tat tta cag gct cag ttg tgt aac ctt cca caa ggt agt cta cag aat 1610

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

400 405 410

aaa act aaa tta ctt cct agt att gag gag gcg tgg agc cta cca att 1658

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

415 420 425

ccc gca gag ctt acc tcc agg cag ggt gcc atg aac aca gca cag cag 1706

Pro Ala Glu Leu Thr Ser Arg Gln Gly Ala Met Asn Thr Ala Gln Gln

430 435 440

aat act tct gac aat tgg agt ggt gga cat gct gtg tca cat cct cca 1754

Asn Thr Ser Asp Asn Trp Ser Gly Gly His Ala Val Ser His Pro Pro

445 450 455

gta cag caa caa gct cat tca tgg tgt ttg aca cca cag aaa tta cag 1802

Val Gln Gln Gln Ala His Ser Trp Cys Leu Thr Pro Gln Lys Leu Gln

460 465 470 475

cat ttg gaa cag ctc cgc gca aat aga aat aat tta aat cca gca cag 1850

His Leu Glu Gln Leu Arg Ala Asn Arg Asn Asn Leu Asn Pro Ala Gln

480 485 490

aaa ctg atg ctg gaa cag ctg gaa agt cag ttt gtc tta atg caa caa 1898

Lys Leu Met Leu Glu Gln Leu Glu Ser Gln Phe Val Leu Met Gln Gln

495 500 505

cac caa atg aga cca aca gga gtt gca cag gta cga tct act gga att 1946

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

510 515 520

cct aat ggg cca aca gct gac tca tca ctg cct aca aac tca gtc tct 1994

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

525 530 535

ggc cag cag cca cag ctt gct ctg acc aga gtg cct agc gtc tct cag 2042

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

540 545 550 555

cct gga gtc cgt cct gcc tgc cct ggg cag cct ttg gcc aat gga ccc 2090

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

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

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aca gac act att ttg ata ggc aat aat cat ata aca gga agt gga agt 2186

Thr Asp Thr Ile Leu Ile Gly Asn Asn His Ile Thr Gly Ser Gly Ser

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aat gga aac gtg cct tac ctg cag cga aac gca ctc act cta cct cat 2234

Asn Gly Asn Val Pro Tyr Leu Gln Arg Asn Ala Leu Thr Leu Pro His

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aac cgc aca aac ctg acc agc agc gca gag gag ccg tgg aaa aac caa 2282

Asn Arg Thr Asn Leu Thr Ser Ser Ala Glu Glu Pro Trp Lys Asn Gln

620 625 630 635

cta tct aac tcc act cag ggg ctt cac aaa ggt cag agt tca cat tcg 2330

Leu Ser Asn Ser Thr Gln Gly Leu His Lys Gly Gln Ser Ser His Ser

640 645 650

gca ggt cct aat ggt gaa cga cct ctc tct tcc act ggg cct tcc cag 2378

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

655 660 665

cat ctc cag gca gct ggc tct ggt att cag aat cag aac gga cat ccc 2426

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

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acc ctg cct agc aat tca gta aca cag ggg gct gct ctc aat cac ctc 2474

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

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tcc tct cac act gct acc tca ggt gga caa caa ggc att acc tta acc 2522

Ser Ser His Thr Ala Thr Ser Gly Gly Gln Gln Gly Ile Thr Leu Thr

700 705 710 715

aaa gag agc aag cct tca gga aac ata ttg acg gtg cct gaa aca agc 2570

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

720 725 730

agg cac act gga gag aca cct aac agc act gcc agt gtc gag gga ctt 2618

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

735 740 745

cct aat cat gtc cat cag atg acg gca gat gct gtt tgc agt cct agc 2666

Pro Asn His Val His Gln Met Thr Ala Asp Ala Val Cys Ser Pro Ser

750 755 760

cat gga gat tct aag tca cca ggt tta cta agt tca gac aat cct cag 2714

His Gly Asp Ser Lys Ser Pro Gly Leu Leu Ser Ser Asp Asn Pro Gln

765 770 775

ctc tct gcc ttg ttg atg gga aaa gcc aat aac aat gtg ggt act gga 2762

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

780 785 790 795

acc tgt gac aaa gtc aat aac atc cac cca gct gtt cat aca aag act 2810

Thr Cys Asp Lys Val Asn Asn Ile His Pro Ala Val His Thr Lys Thr

800 805 810

gat aac tct gtt gcc tct tca cca tct tca gcc att tca aca gca aca 2858

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

815 820 825

cct tct cca aaa tcc act gag cag aca acc aca aac agt gtt acc agc 2906

Pro Ser Pro Lys Ser Thr Glu Gln Thr Thr Thr Asn Ser Val Thr Ser

830 835 840

ctt aac agc cct cac agt ggg cta cac aca att aat gga gaa ggg atg 2954

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

845 850 855

gaa gaa tct cag agc ccc atg aaa aca gat ctg ctt ctg gtt aac cac 3002

Glu Glu Ser Gln Ser Pro Met Lys Thr Asp Leu Leu Leu Val Asn His

860 865 870 875

aaa cct agt cca cag atc ata cca tca atg tct gtg tcc ata tac ccc 3050

Lys Pro Ser Pro Gln Ile Ile Pro Ser Met Ser Val Ser Ile Tyr Pro

880 885 890

agc tca gca gaa gtt ctg aag gca tgc agg aat cta ggt aaa aat ggc 3098

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

895 900 905

tta tct aac agt agc att ttg ttg gat aaa tgt cca cct cca aga cca 3146

Leu Ser Asn Ser Ser Ile Leu Leu Asp Lys Cys Pro Pro Pro Arg Pro

910 915 920

cca tct tca cca tac cct ccc ttg cca aag gac aag ttg aat cca cct 3194

Pro Ser Ser Pro Tyr Pro Pro Leu Pro Lys Asp Lys Leu Asn Pro Pro

925 930 935

aca cct agt att tac ttg gaa aat aaa cgt gat gct ttc ttt cct cca 3242

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

940 945 950 955

tta cat caa ttt tgt aca aat ccg aac aac cct gtt aca gta ata cgt 3290

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

960 965 970

ggc ctt gct gga gct ctt aag tta gac ctg gga ctt ttc tct act aaa 3338

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

975 980 985

act ttg gtg gaa gct aac aat gaa cat atg gta gaa gtg agg aca cag 3386

Thr Leu Val Glu Ala Asn Asn Glu His Met Val Glu Val Arg Thr Gln

990 995 1000

ttg ttg cag cca gca gat gaa aac tgg gat ccc act gga aca aag 3431

Leu Leu Gln Pro Ala Asp Glu Asn Trp Asp Pro Thr Gly Thr Lys

1005 1010 1015

aaa atc tgg cat tgt gaa agt aat aga tct cat act aca att gct 3476

Lys Ile Trp His Cys Glu Ser Asn Arg Ser His Thr Thr Ile Ala

1020 1025 1030

aaa tat gca cag tac cag gcc tcc tca ttc cag gaa tca ttg aga 3521

Lys Tyr Ala Gln Tyr Gln Ala Ser Ser Phe Gln Glu Ser Leu Arg

1035 1040 1045

gaa gaa aat gaa aaa aga agt cat cat aaa gac cac tca gat agt 3566

Glu Glu Asn Glu Lys Arg Ser His His Lys Asp His Ser Asp Ser

1050 1055 1060

gaa tct aca tcg tca gat aat tct ggg agg agg agg aaa gga ccc 3611

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

1065 1070 1075

ttt aaa acc ata aag ttt ggg acc aat att gac cta tct gat gac 3656

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

1080 1085 1090

aaa aag tgg aag ttg cag cta cat gag ctg act aaa ctt cct gct 3701

Lys Lys Trp Lys Leu Gln Leu His Glu Leu Thr Lys Leu Pro Ala

1095 1100 1105

ttt gtg cgt gtc gta tca gca gga aat ctt cta agc cat gtt ggt 3746

Phe Val Arg Val Val Ser Ala Gly Asn Leu Leu Ser His Val Gly

1110 1115 1120

cat acc ata ttg ggc atg aac aca gtt caa cta tac atg aaa gtt 3791

His Thr Ile Leu Gly Met Asn Thr Val Gln Leu Tyr Met Lys Val

1125 1130 1135

cca ggg agc aga aca cca ggt cat cag gaa aat aac aac ttc tgt 3836

Pro Gly Ser Arg Thr Pro Gly His Gln Glu Asn Asn Asn Phe Cys

1140 1145 1150

tca gtt aac ata aat att ggc cca ggt gac tgt gaa tgg ttt gtt 3881

Ser Val Asn Ile Asn Ile Gly Pro Gly Asp Cys Glu Trp Phe Val

1155 1160 1165

gtt cct gaa ggt tac tgg ggt gtt ctg aat gac ttc tgt gaa aaa 3926

Val Pro Glu Gly Tyr Trp Gly Val Leu Asn Asp Phe Cys Glu Lys

1170 1175 1180

aat aat ttg aat ttc cta atg ggt tct tgg tgg ccc aat ctt gaa 3971

Asn Asn Leu Asn Phe Leu Met Gly Ser Trp Trp Pro Asn Leu Glu

1185 1190 1195

gat ctt tat gaa gca aat gtt cca gtg tat agg ttt att cag cga 4016

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

1200 1205 1210

cct gga gat ttg gtc tgg ata aat gca ggc act gtt cat tgg gtt 4061

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

1215 1220 1225

cag gct att ggc tgg tgc aac aac att gct tgg aat gtt ggt cca 4106

Gln Ala Ile Gly Trp Cys Asn Asn Ile Ala Trp Asn Val Gly Pro

1230 1235 1240

ctt aca gcc tgc cag tat aaa ttg gca gtg gaa cgg tac gaa tgg 4151

Leu Thr Ala Cys Gln Tyr Lys Leu Ala Val Glu Arg Tyr Glu Trp

1245 1250 1255

aac aaa ttg caa agt gtg aag tca ata gta ccc atg gtt cat ctt 4196

Asn Lys Leu Gln Ser Val Lys Ser Ile Val Pro Met Val His Leu

1260 1265 1270

tcc tgg aat atg gca cga aat atc aag gtc tca gat cca aag ctt 4241

Ser Trp Asn Met Ala Arg Asn Ile Lys Val Ser Asp Pro Lys Leu

1275 1280 1285

ttt gaa atg att aag tat tgt ctt cta aga act ctg aag caa tgt 4286

Phe Glu Met Ile Lys Tyr Cys Leu Leu Arg Thr Leu Lys Gln Cys

1290 1295 1300

cag aca ttg agg gaa gct ctc att gct gca gga aaa gag att ata 4331

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

1305 1310 1315

tgg cat ggg cgg aca aaa gaa gaa cca gct cat tac tgt agc att 4376

Trp His Gly Arg Thr Lys Glu Glu Pro Ala His Tyr Cys Ser Ile

1320 1325 1330

tgt gaa gtg gag gtt ttt gat ctg ctt ttt gtc act aat gag agt 4421

Cys Glu Val Glu Val Phe Asp Leu Leu Phe Val Thr Asn Glu Ser

1335 1340 1345

aat tca cga aag acc tac ata gta cat tgc caa gat tgt gca cga 4466

Asn Ser Arg Lys Thr Tyr Ile Val His Cys Gln Asp Cys Ala Arg

1350 1355 1360

aaa aca agc gga aac ttg gaa aac ttt gtg gtg cta gaa cag tac 4511

Lys Thr Ser Gly Asn Leu Glu Asn Phe Val Val Leu Glu Gln Tyr

1365 1370 1375

aaa atg gag gac ctg atg caa gtc tat gac caa ttt aca tta gct 4556

Lys Met Glu Asp Leu Met Gln Val Tyr Asp Gln Phe Thr Leu Ala

1380 1385 1390

cct cca tta cca tcc gcc tca tct tga tattgttcca tggacattaa 4603

Pro Pro Leu Pro Ser Ala Ser Ser

1395 1400

atgagacctt ttctgctatt caggaaataa cccagttctg caccactggt ttttgtagct 4663

atctcgtaag gctgctggct gaaaactgtg tctatgcaac cttccaagtg cggagtgtca 4723

accaactgga cgggagagag tactgctcct actccaggac tctcacaaag ctgatgagct 4783

gtacttcaga aaaaaataat aatttccatg ttttgtatat atctgacaaa actggcaaca 4843

tcttacagac tactgacttg aagacaacct cttttatatt tctctatttc tgggctgatg 4903

aatttgtttt catctgtctt ttcccccttc agaattttcc ttggaaaaaa aatactagcc 4963

tagctggtca tttctttgta aggtagttag caattttaag tctttctttg gtcaactttt 5023

ttttaatgtg aaaagttagg taagacactt ttttactgct tttatgtttt tctgtcttgt 5083

tttgagacca tgatggttac acttttggtt cctaaataaa atttaaaaaa ttaacagcca 5143

agtcacaaag gtaatggatt gcacatagac taaggaataa acttcagatt tgtgattttt 5203

gtttctaatc ttgatgtaaa tttacactat ttataaatac atatttattg cttgaaaata 5263

tttgtgaatg gaatgctgtt attttttcca gatttacctg ccattgaaat tttaaggagt 5323

tctgtaattt caaacactac tcctattaca ttttctatgt gtaaataaaa ctgcttagca 5383

ttgtacagaa acttttatta aaattgttta atgtttaaag agttttctat tgtttgagtt 5443

ttaaaaaaga ctttatgtac agtgcccagt ttttgttcat ttttgaaatc tgattatata 5503

tattttatat atacttatgt atgtatatat aatatatata gaaatctgga tatatatgta 5563

taaatcttta gaacttaaat ttttctcgtt ttaagtttca catctatggt agatttttga 5623

ggtgtctact gtaaagtatt gcttacaaaa agtatgatta tttttaaaga aatatatatg 5683

gtatgtatcc tcaagaccta aaatgtcaga ctggtttatt gttaagttgc aattactgca 5743

atgacagacc aataaacaat tgctgccaaa atgtagtata aa 5785

<210> 2

<211> 1401

<212> PRT

<213>Homo sapiens (Homo sapiens)

<400> 2

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

1 5 10 15

Ala Phe Gly Asp Glu Glu Lys Lys Met Ala Ala Gly Lys Ala Ser Gly

20 25 30

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

35 40 45

Leu Gly Gly Leu Asp Ser Arg Leu Phe Gly Phe Val Arg Phe His Glu

50 55 60

Asp Gly Ala Arg Thr Lys Ala Leu Leu Gly Lys Ala Val Arg Cys Tyr

65 70 75 80

Glu Ser Leu Ile Leu Lys Ala Glu Gly Lys Val Glu Ser Asp Phe Phe

85 90 95

Cys Gln Leu Gly His Phe Asn Leu Leu Leu Glu Asp Tyr Pro Lys Ala

100 105 110

Leu Ser Ala Tyr Gln Arg Tyr Tyr Ser Leu Gln Ser Asp Tyr Trp Lys

115 120 125

Asn Ala Ala Phe Leu Tyr Gly Leu Gly Leu Val Tyr Phe His Tyr Asn

130 135 140

Ala Phe Gln Trp Ala Ile Lys Ala Phe Gln Glu Val Leu Tyr Val Asp

145 150 155 160

Pro Ser Phe Cys Arg Ala Lys Glu Ile His Leu Arg Leu Gly Leu Met

165 170 175

Phe Lys Val Asn Thr Asp Tyr Glu Ser Ser Leu Lys His Phe Gln Leu

180 185 190

Ala Leu Val Asp Cys Asn Pro Cys Thr Leu Ser Asn Ala Glu Ile Gln

195 200 205

Phe His Ile Ala His Leu Tyr Glu Thr Gln Arg Lys Tyr His Ser Ala

210 215 220

Lys Glu Ala Tyr Glu Gln Leu Leu Gln Thr Glu Asn Leu Ser Ala Gln

225 230 235 240

Val Lys Ala Thr Val Leu Gln Gln Leu Gly Trp Met His His Thr Val

245 250 255

Asp Leu Leu Gly Asp Lys Ala Thr Lys Glu Ser Tyr Ala Ile Gln Tyr

260 265 270

Leu Gln Lys Ser Leu Glu Ala Asp Pro Asn Ser Gly Gln Ser Trp Tyr

275 280 285

Phe Leu Gly Arg Cys Tyr Ser Ser Ile Gly Lys Val Gln Asp Ala Phe

290 295 300

Ile Ser Tyr Arg Gln Ser Ile Asp Lys Ser Glu Ala Ser Ala Asp Thr

305 310 315 320

Trp Cys Ser Ile Gly Val Leu Tyr Gln Gln Gln Asn Gln Pro Met Asp

325 330 335

Ala Leu Gln Ala Tyr Ile Cys Ala Val Gln Leu Asp His Gly His Ala

340 345 350

Ala Ala Trp Met Asp Leu Gly Thr Leu Tyr Glu Ser Cys Asn Gln Pro

355 360 365

Gln Asp Ala Ile Lys Cys Tyr Leu Asn Ala Thr Arg Ser Lys Ser Cys

370 375 380

Ser Asn Thr Ser Ala Leu Ala Ala Arg Ile Lys Tyr Leu Gln Ala Gln

385 390 395 400

Leu Cys Asn Leu Pro Gln Gly Ser Leu Gln Asn Lys Thr Lys Leu Leu

405 410 415

Pro Ser Ile Glu Glu Ala Trp Ser Leu Pro Ile Pro Ala Glu Leu Thr

420 425 430

Ser Arg Gln Gly Ala Met Asn Thr Ala Gln Gln Asn Thr Ser Asp Asn

435 440 445

Trp Ser Gly Gly His Ala Val Ser His Pro Pro Val Gln Gln Gln Ala

450 455 460

His Ser Trp Cys Leu Thr Pro Gln Lys Leu Gln His Leu Glu Gln Leu

465 470 475 480

Arg Ala Asn Arg Asn Asn Leu Asn Pro Ala Gln Lys Leu Met Leu Glu

485 490 495

Gln Leu Glu Ser Gln Phe Val Leu Met Gln Gln His Gln Met Arg Pro

500 505 510

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

515 520 525

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

530 535 540

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

545 550 555 560

Ala Cys Pro Gly Gln Pro Leu Ala Asn Gly Pro Phe Ser Ala Gly His

565 570 575

Val Pro Cys Ser Thr Ser Arg Thr Leu Gly Ser Thr Asp Thr Ile Leu

580 585 590

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

595 600 605

Tyr Leu Gln Arg Asn Ala Leu Thr Leu Pro His Asn Arg Thr Asn Leu

610 615 620

Thr Ser Ser Ala Glu Glu Pro Trp Lys Asn Gln Leu Ser Asn Ser Thr

625 630 635 640

Gln Gly Leu His Lys Gly Gln Ser Ser His Ser Ala Gly Pro Asn Gly

645 650 655

Glu Arg Pro Leu Ser Ser Thr Gly Pro Ser Gln His Leu Gln Ala Ala

660 665 670

Gly Ser Gly Ile Gln Asn Gln Asn Gly His Pro Thr Leu Pro Ser Asn

675 680 685

Ser Val Thr Gln Gly Ala Ala Leu Asn His Leu Ser Ser His Thr Ala

690 695 700

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

705 710 715 720

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

725 730 735

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

740 745 750

Gln Met Thr Ala Asp Ala Val Cys Ser Pro Ser His Gly Asp Ser Lys

755 760 765

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

770 775 780

Met Gly Lys Ala Asn Asn Asn Val Gly Thr Gly Thr Cys Asp Lys Val

785 790 795 800

Asn Asn Ile His Pro Ala Val His Thr Lys Thr Asp Asn Ser Val Ala

805 810 815

Ser Ser Pro Ser Ser Ala Ile Ser Thr Ala Thr Pro Ser Pro Lys Ser

820 825 830

Thr Glu Gln Thr Thr Thr Asn Ser Val Thr Ser Leu Asn Ser Pro His

835 840 845

Ser Gly Leu His Thr Ile Asn Gly Glu Gly Met Glu Glu Ser Gln Ser

850 855 860

Pro Met Lys Thr Asp Leu Leu Leu Val Asn His Lys Pro Ser Pro Gln

865 870 875 880

Ile Ile Pro Ser Met Ser Val Ser Ile Tyr Pro Ser Ser Ala Glu Val

885 890 895

Leu Lys Ala Cys Arg Asn Leu Gly Lys Asn Gly Leu Ser Asn Ser Ser

900 905 910

Ile Leu Leu Asp Lys Cys Pro Pro Pro Arg Pro Pro Ser Ser Pro Tyr

915 920 925

Pro Pro Leu Pro Lys Asp Lys Leu Asn Pro Pro Thr Pro Ser Ile Tyr

930 935 940

Leu Glu Asn Lys Arg Asp Ala Phe Phe Pro Pro Leu His Gln Phe Cys

945 950 955 960

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

965 970 975

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

980 985 990

Asn Asn Glu His Met Val Glu Val Arg Thr Gln Leu Leu Gln Pro Ala

995 1000 1005

Asp Glu Asn Trp Asp Pro Thr Gly Thr Lys Lys Ile Trp His Cys

1010 1015 1020

Glu Ser Asn Arg Ser His Thr Thr Ile Ala Lys Tyr Ala Gln Tyr

1025 1030 1035

Gln Ala Ser Ser Phe Gln Glu Ser Leu Arg Glu Glu Asn Glu Lys

1040 1045 1050

Arg Ser His His Lys Asp His Ser Asp Ser Glu Ser Thr Ser Ser

1055 1060 1065

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

1070 1075 1080

Phe Gly Thr Asn Ile Asp Leu Ser Asp Asp Lys Lys Trp Lys Leu

1085 1090 1095

Gln Leu His Glu Leu Thr Lys Leu Pro Ala Phe Val Arg Val Val

1100 1105 1110

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

1115 1120 1125

Met Asn Thr Val Gln Leu Tyr Met Lys Val Pro Gly Ser Arg Thr

1130 1135 1140

Pro Gly His Gln Glu Asn Asn Asn Phe Cys Ser Val Asn Ile Asn

1145 1150 1155

Ile Gly Pro Gly Asp Cys Glu Trp Phe Val Val Pro Glu Gly Tyr

1160 1165 1170

Trp Gly Val Leu Asn Asp Phe Cys Glu Lys Asn Asn Leu Asn Phe

1175 1180 1185

Leu Met Gly Ser Trp Trp Pro Asn Leu Glu Asp Leu Tyr Glu Ala

1190 1195 1200

Asn Val Pro Val Tyr Arg Phe Ile Gln Arg Pro Gly Asp Leu Val

1205 1210 1215

Trp Ile Asn Ala Gly Thr Val His Trp Val Gln Ala Ile Gly Trp

1220 1225 1230

Cys Asn Asn Ile Ala Trp Asn Val Gly Pro Leu Thr Ala Cys Gln

1235 1240 1245

Tyr Lys Leu Ala Val Glu Arg Tyr Glu Trp Asn Lys Leu Gln Ser

1250 1255 1260

Val Lys Ser Ile Val Pro Met Val His Leu Ser Trp Asn Met Ala

1265 1270 1275

Arg Asn Ile Lys Val Ser Asp Pro Lys Leu Phe Glu Met Ile Lys

1280 1285 1290

Tyr Cys Leu Leu Arg Thr Leu Lys Gln Cys Gln Thr Leu Arg Glu

1295 1300 1305

Ala Leu Ile Ala Ala Gly Lys Glu Ile Ile Trp His Gly Arg Thr

1310 1315 1320

Lys Glu Glu Pro Ala His Tyr Cys Ser Ile Cys Glu Val Glu Val

1325 1330 1335

Phe Asp Leu Leu Phe Val Thr Asn Glu Ser Asn Ser Arg Lys Thr

1340 1345 1350

Tyr Ile Val His Cys Gln Asp Cys Ala Arg Lys Thr Ser Gly Asn

1355 1360 1365

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

1370 1375 1380

Met Gln Val Tyr Asp Gln Phe Thr Leu Ala Pro Pro Leu Pro Ser

1385 1390 1395

Ala Ser Ser

1400

<210> 3

<211> 5918

<212> DNA

<213>Mouse (Mus musculus)

<220>

<221> CDS

<222> (432)..(4637)

<400> 3

ggctgcgccg agcgggctgt gagggggggg tcacggcagc ggcgtggggt tcgctgtgtg 60

acacaattac aacaactttg tgctggtgcc ggggaagttt gtgtctccta cgaatccccc 120

ccagagctcc cgctcctcgc gcgctccggc cattcccgcc gaccctgcct gtggctgccc 180

ccttcccatc cccgcgatgt gaccctacag ccgagccgtc gccgctccgg acccagtggt 240

tccacagcct ctgctgcctc tgccttcacc tctaccgcgt taggattttt cgctgccgcc 300

tctcgcggca gaggaggagg cagaaataaa gttggtgtgc cggtcttgtg cggagattgg 360

aggcgtcctt gcgggctccg gcccggagag ggggggtgtc ggcgttggag ttgtgaattc 420

gctgcgtttc c atg aaa tcc tgc gga gtg tcg ctc gct acc gcc gcc gcc 470

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

1 5 10

gcc gcc gcc gcc gcc gct ttc ggt gat gag gaa aag aaa atg gcg gcg 518

Ala Ala Ala Ala Ala Ala Phe Gly Asp Glu Glu Lys Lys Met Ala Ala

15 20 25

gga aaa gcg agc ggc gag agc gag gag gcg tcc ccc agc ctg aca gcg 566

Gly Lys Ala Ser Gly Glu Ser Glu Glu Ala Ser Pro Ser Leu Thr Ala

30 35 40 45

gag gag agg gag gcg ctc ggc gga ctg gac agc cgc ctt ttc ggg ttc 614

Glu Glu Arg Glu Ala Leu Gly Gly Leu Asp Ser Arg Leu Phe Gly Phe

50 55 60

gtg agg ttt cat gaa gat ggc gcc agg atg aag gcc ctg ctg ggc aag 662

Val Arg Phe His Glu Asp Gly Ala Arg Met Lys Ala Leu Leu Gly Lys

65 70 75

gct gtt cgc tgc tac gaa tct cta atc tta aaa gct gaa ggg aaa gtg 710

Ala Val Arg Cys Tyr Glu Ser Leu Ile Leu Lys Ala Glu Gly Lys Val

80 85 90

gag tct gat ttc ttt tgt caa tta ggt cac ttc aac ctc tta ttg gaa 758

Glu Ser Asp Phe Phe Cys Gln Leu Gly His Phe Asn Leu Leu Leu Glu

95 100 105

gat tat cca aaa gca tta tct gca tac cag agg tac tac agt tta cag 806

Asp Tyr Pro Lys Ala Leu Ser Ala Tyr Gln Arg Tyr Tyr Ser Leu Gln

110 115 120 125

tct gat tac tgg aag aat gct gcc ttt tta tat ggt ctt ggt ttg gtc 854

Ser Asp Tyr Trp Lys Asn Ala Ala Phe Leu Tyr Gly Leu Gly Leu Val

130 135 140

tac ttc cat tac aat gca ttt cag tgg gct att aaa gca ttt cag gag 902

Tyr Phe His Tyr Asn Ala Phe Gln Trp Ala Ile Lys Ala Phe Gln Glu

145 150 155

gtg ctt tat gtc gat ccc agc ttt tgt cga gcc aag gaa att cat tta 950

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

160 165 170

cga ctt ggg ctt atg ttc aaa gtg aac aca gac tat gag tct agt tta 998

Arg Leu Gly Leu Met Phe Lys Val Asn Thr Asp Tyr Glu Ser Ser Leu

175 180 185

aag cat ttt cag tta gct ttg gtt gac tgt aat ccc tgc act ttg tcc 1046

Lys His Phe Gln Leu Ala Leu Val Asp Cys Asn Pro Cys Thr Leu Ser

190 195 200 205

aat gct gaa att cag ttt cac att gcc cac tta tat gaa acc cag agg 1094

Asn Ala Glu Ile Gln Phe His Ile Ala His Leu Tyr Glu Thr Gln Arg

210 215 220

aag tat cat tct gca aaa gaa gct tat gag caa ctt ttg cag aca gaa 1142

Lys Tyr His Ser Ala Lys Glu Ala Tyr Glu Gln Leu Leu Gln Thr Glu

225 230 235

aac ctt tct gca caa gta aaa gca act att tta caa caa tta ggt tgg 1190

Asn Leu Ser Ala Gln Val Lys Ala Thr Ile Leu Gln Gln Leu Gly Trp

240 245 250

atg cat cac act gtg gat ctc ctg gga gat aag gcc acc aag gaa agt 1238

Met His His Thr Val Asp Leu Leu Gly Asp Lys Ala Thr Lys Glu Ser

255 260 265

tat gct att cag tat ctc cag aag tcc ttg gaa gca gat cca aat tct 1286

Tyr Ala Ile Gln Tyr Leu Gln Lys Ser Leu Glu Ala Asp Pro Asn Ser

270 275 280 285

ggc cag tcc tgg tat ttc ctt gga agg tgc tat tca agt att ggg aaa 1334

Gly Gln Ser Trp Tyr Phe Leu Gly Arg Cys Tyr Ser Ser Ile Gly Lys

290 295 300

gtt cag gat gcc ttt ata tct tac agg caa tct att gat aaa tca gaa 1382

Val Gln Asp Ala Phe Ile Ser Tyr Arg Gln Ser Ile Asp Lys Ser Glu

305 310 315

gca agt gca gat aca tgg tgt tca ata ggt gtg ctc tat caa cag caa 1430

Ala Ser Ala Asp Thr Trp Cys Ser Ile Gly Val Leu Tyr Gln Gln Gln

320 325 330

aat cag cct atg gat gct ttg caa gct tat att tgt gct gta caa ttg 1478

Asn Gln Pro Met Asp Ala Leu Gln Ala Tyr Ile Cys Ala Val Gln Leu

335 340 345

gac cac ggt cat gct gca gcc tgg atg gat cta ggc act ctc tat gaa 1526

Asp His Gly His Ala Ala Ala Trp Met Asp Leu Gly Thr Leu Tyr Glu

350 355 360 365

tcc tgc aac caa cct cag gat gct att aaa tgc tat tta aat gca act 1574

Ser Cys Asn Gln Pro Gln Asp Ala Ile Lys Cys Tyr Leu Asn Ala Thr

370 375 380

aga agc aaa aat tgt agt aat acc tct gga ctt gca gca cga att aag 1622

Arg Ser Lys Asn Cys Ser Asn Thr Ser Gly Leu Ala Ala Arg Ile Lys

385 390 395

tat tta cag gct cag ttg tgt aac ctt cca caa ggt agt cta cag aat 1670

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

400 405 410

aaa act aaa tta ctt cct agt att gag gag gca tgg agc cta cca atc 1718

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

415 420 425

ccc gca gag ctt acc tcc agg cag ggt gcc atg aac aca gca cag cag 1766

Pro Ala Glu Leu Thr Ser Arg Gln Gly Ala Met Asn Thr Ala Gln Gln

430 435 440 445

aat act tct gat aat tgg agt ggt ggc aat gca cca cct cca gta gaa 1814

Asn Thr Ser Asp Asn Trp Ser Gly Gly Asn Ala Pro Pro Pro Val Glu

450 455 460

caa caa act cat tca tgg tgt ttg aca cca cag aaa tta cag cac ttg 1862

Gln Gln Thr His Ser Trp Cys Leu Thr Pro Gln Lys Leu Gln His Leu

465 470 475

gaa cag ctc cga gca aac aga aat aat tta aat cca gca cag aaa cta 1910

Glu Gln Leu Arg Ala Asn Arg Asn Asn Leu Asn Pro Ala Gln Lys Leu

480 485 490

atg ctg gaa cag ctg gaa agt cag ttt gtc tta atg cag caa cac caa 1958

Met Leu Glu Gln Leu Glu Ser Gln Phe Val Leu Met Gln Gln His Gln

495 500 505

atg aga caa aca gga gtt gca cag gta cgg cct act gga att ctt aat 2006

Met Arg Gln Thr Gly Val Ala Gln Val Arg Pro Thr Gly Ile Leu Asn

510 515 520 525

ggg cca aca gtt gac tca tca ctg cct aca aac tca gtt tct ggc cag 2054

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

530 535 540

cag cca cag ctt cct ctg acc aga atg cct agt gtc tct cag cct gga 2102

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

545 550 555

gtc cac act gcc tgt cct agg cag act ttg gcc aat gga ccc ttt tct 2150

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

560 565 570

gca ggc cat gtt ccc tgt agc aca tca aga aca ctg gga agt aca gac 2198

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

575 580 585

act gtt ttg ata ggc aat aat cat gta aca gga agt gga agt aat gga 2246

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

590 595 600 605

aac gtg cct tac ctg cag cga aac gca ccc act cta cct cat aac cgc 2294

Asn Val Pro Tyr Leu Gln Arg Asn Ala Pro Thr Leu Pro His Asn Arg

610 615 620

aca aac ctg acc agc agc aca gag gag ccg tgg aaa aac caa cta tct 2342

Thr Asn Leu Thr Ser Ser Thr Glu Glu Pro Trp Lys Asn Gln Leu Ser

625 630 635

aac tcc act cag ggg ctt cac aaa ggt ccg agt tca cat ttg gca ggt 2390

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

640 645 650

cct aat ggt gaa cga cct cta tct tcc act ggg cct tcc cag cat ctc 2438

Pro Asn Gly Glu Arg Pro Leu Ser Ser Thr Gly Pro Ser Gln His Leu

655 660 665

cag gca gct ggc tct ggt att cag aat cag aat gga cat ccc acc ctg 2486

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

670 675 680 685

cct agc aat tca gta aca cag ggg gct gct ctc aat cac ctc tcc tct 2534

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

690 695 700

cac act gct acc tca ggt gga caa caa ggc att acc tta acc aaa gag 2582

His Thr Ala Thr Ser Gly Gly Gln Gln Gly Ile Thr Leu Thr Lys Glu

705 710 715

agc aag cct tca gga aac aca ttg acg gtg cct gaa aca agc agg caa 2630

Ser Lys Pro Ser Gly Asn Thr Leu Thr Val Pro Glu Thr Ser Arg Gln

720 725 730

act gga gag aca cct aac agc act gcc agt gtt gag gga ctt cct aat 2678

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

735 740 745

cat gtc cat cag gtg atg gca gat gct gtt tgc agt cct agc cat gga 2726

His Val His Gln Val Met Ala Asp Ala Val Cys Ser Pro Ser His Gly

750 755 760 765

gat tct aag tca cca ggt tta cta agt tca gac aat cct cag ctc tct 2774

Asp Ser Lys Ser Pro Gly Leu Leu Ser Ser Asp Asn Pro Gln Leu Ser

770 775 780

gcc ttg ttg atg gga aaa gct aat aac aat gtg ggt cct gga acc tgt 2822

Ala Leu Leu Met Gly Lys Ala Asn Asn Asn Val Gly Pro Gly Thr Cys

785 790 795

gac aaa gtc aat aac atc cac cca act gtc cat aca aag act gat aat 2870

Asp Lys Val Asn Asn Ile His Pro Thr Val His Thr Lys Thr Asp Asn

800 805 810

tct gtt gcc tct tca cca tct tca gcc att tcc aca gca aca cct tct 2918

Ser Val Ala Ser Ser Pro Ser Ser Ala Ile Ser Thr Ala Thr Pro Ser

815 820 825

cct aag tcc act gaa cag aca acc aca aac agt gtt acc agc ctt aac 2966

Pro Lys Ser Thr Glu Gln Thr Thr Thr Asn Ser Val Thr Ser Leu Asn

830 835 840 845

agc cct cac agt ggg ctg cac aca att aat gga gaa gga atg gaa gaa 3014

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

850 855 860

tct cag agc ccc att aaa aca gat ctg ctt cta gtt agc cac aga cct 3062

Ser Gln Ser Pro Ile Lys Thr Asp Leu Leu Leu Val Ser His Arg Pro

865 870 875

agt cct cag atc ata cca tca atg tct gtg tcc ata tat ccc agc tca 3110

Ser Pro Gln Ile Ile Pro Ser Met Ser Val Ser Ile Tyr Pro Ser Ser

880 885 890

gca gaa gtt ctg aaa gct tgc agg aat cta ggt aaa aac ggc ctg tct 3158

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

895 900 905

aat agt agc att ctg ttg gat aaa tgt ccg cct cca aga cca cca tcc 3206

Asn Ser Ser Ile Leu Leu Asp Lys Cys Pro Pro Pro Arg Pro Pro Ser

910 915 920 925

tca cca tac cct ccc ttg cca aag gac aag ttg aat cca cct aca cct 3254

Ser Pro Tyr Pro Pro Leu Pro Lys Asp Lys Leu Asn Pro Pro Thr Pro

930 935 940

agt att tat ttg gaa aat aaa cgt gat gct ttc ttt cct cca tta cat 3302

Ser Ile Tyr Leu Glu Asn Lys Arg Asp Ala Phe Phe Pro Pro Leu His

945 950 955

caa ttt tgt aca aac cca aac aac cct gtt aca gta ata cgt ggc ctt 3350

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

960 965 970

gct gga gct ctt aaa tta gac ttg gga ctt ttc tct act aaa act ttg 3398

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

975 980 985

gtg gaa gct aac aat gaa cat atg gta gaa gtg agg aca cag ttg tta 3446

Val Glu Ala Asn Asn Glu His Met Val Glu Val Arg Thr Gln Leu Leu

990 995 1000 1005

caa cca gca gat gaa aat tgg gac cct act gga acc aag aaa atc 3491

Gln Pro Ala Asp Glu Asn Trp Asp Pro Thr Gly Thr Lys Lys Ile

1010 1015 1020

tgg cac tgt gaa agt aat aga tct cat act aca att gct aaa tat 3536

Trp His Cys Glu Ser Asn Arg Ser His Thr Thr Ile Ala Lys Tyr

1025 1030 1035

gct cag tac cag gcc tcc tca ttc caa gaa tca ttg aga gaa gaa 3581

Ala Gln Tyr Gln Ala Ser Ser Phe Gln Glu Ser Leu Arg Glu Glu

1040 1045 1050

aat gag aaa aga agt cac cat aaa gac cac tca gac agt gaa tct 3626

Asn Glu Lys Arg Ser His His Lys Asp His Ser Asp Ser Glu Ser

1055 1060 1065

aca tca tca gat aat tct ggg aaa aga aga aaa gga ccc ttt aaa 3671

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

1070 1075 1080

acc att aag ttt ggg acc aac att gac ctg tct gat gac aaa aag 3716

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

1085 1090 1095

tgg aag tta cag cta cat gag ctg act aaa ctt cct gcc ttc gtg 3761

Trp Lys Leu Gln Leu His Glu Leu Thr Lys Leu Pro Ala Phe Val

1100 1105 1110

aga gtt gta tct gca gga aat ctt tta agc cac gtt ggt cat act 3806

Arg Val Val Ser Ala Gly Asn Leu Leu Ser His Val Gly His Thr

1115 1120 1125

ata ctg ggc atg aac aca gtt caa cta tac atg aaa gtt cca gga 3851

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

1130 1135 1140

agc aga aca cca ggt cat caa gaa aat aac aac ttc tgt tca gtt 3896

Ser Arg Thr Pro Gly His Gln Glu Asn Asn Asn Phe Cys Ser Val

1145 1150 1155

aat ata aat att ggc cca ggt gac tgt gaa tgg ttt gtt gtt cct 3941

Asn Ile Asn Ile Gly Pro Gly Asp Cys Glu Trp Phe Val Val Pro

1160 1165 1170

gaa ggc tac tgg ggt gtt ttg aat gac ttc tgt gaa aaa aat aat 3986

Glu Gly Tyr Trp Gly Val Leu Asn Asp Phe Cys Glu Lys Asn Asn

1175 1180 1185

ttg aat ttc tta atg ggt tct tgg tgg ccc aac ctt gaa gat cta 4031

Leu Asn Phe Leu Met Gly Ser Trp Trp Pro Asn Leu Glu Asp Leu

1190 1195 1200

tat gaa gca aat gtt cca gtg tat agg ttt att cag cga cct gga 4076

Tyr Glu Ala Asn Val Pro Val Tyr Arg Phe Ile Gln Arg Pro Gly

1205 1210 1215

gat ctg gtc tgg ata aat gct ggc act gtt cat tgg gtt caa gct 4121

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

1220 1225 1230

att ggc tgg tgc aac aat att gct tgg aat gtt ggt cca ctt aca 4166

Ile Gly Trp Cys Asn Asn Ile Ala Trp Asn Val Gly Pro Leu Thr

1235 1240 1245

gcc tgt cag tat aag tta gca gtg gaa cgt tat gaa tgg aac aag 4211

Ala Cys Gln Tyr Lys Leu Ala Val Glu Arg Tyr Glu Trp Asn Lys

1250 1255 1260

ttg caa aat gta aag tca ata gta ccc atg gtt cat ctt tcc tgg 4256

Leu Gln Asn Val Lys Ser Ile Val Pro Met Val His Leu Ser Trp

1265 1270 1275

aat atg gca cga aat atc aag gtt tca gat cca aag ctt ttt gaa 4301

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

1280 1285 1290

atg att aag tat tgt ctt ctg aga acg ctg aag caa tgt cag aca 4346

Met Ile Lys Tyr Cys Leu Leu Arg Thr Leu Lys Gln Cys Gln Thr

1295 1300 1305

ttg agg gaa gct cta att gct gca gga aaa gag atc ata tgg cac 4391

Leu Arg Glu Ala Leu Ile Ala Ala Gly Lys Glu Ile Ile Trp His

1310 1315 1320

ggg cgg aca aaa gaa gaa cca gct cat tat tgt agt att tgt gag 4436

Gly Arg Thr Lys Glu Glu Pro Ala His Tyr Cys Ser Ile Cys Glu

1325 1330 1335

gtg gag gtt ttt gat ctg ctt ttt gtc act aat gag agt aat tct 4481

Val Glu Val Phe Asp Leu Leu Phe Val Thr Asn Glu Ser Asn Ser

1340 1345 1350

cga aaa acc tac ata gta cat tgc caa gat tgt gca cga aaa aca 4526

Arg Lys Thr Tyr Ile Val His Cys Gln Asp Cys Ala Arg Lys Thr

1355 1360 1365

agt ggg aat ctg gaa aat ttt gtg gtg cta gaa cag tac aaa atg 4571

Ser Gly Asn Leu Glu Asn Phe Val Val Leu Glu Gln Tyr Lys Met

1370 1375 1380

gag gat ctg atg caa gtc tat gac caa ttt aca tta gct cct cca 4616

Glu Asp Leu Met Gln Val Tyr Asp Gln Phe Thr Leu Ala Pro Pro

1385 1390 1395

tta cca tcc gcc tca tct tga tattgttcca tggacattaa acatgagacc 4667

Leu Pro Ser Ala Ser Ser

1400

ttttctgcta ttcagaaagt aacccagttc tgcaccactg ctatttgtag ctatctcgta 4727

aggctgctgg ctgaaaactg tgtctatgca accttccaag tgtggagtgt caaccaactg 4787

gacgggagag tactgctcct atgccaggac tctcgtaaag ctaattagct gaacttcaga 4847

gaaaaaaaaa agaataataa ttaccacgtt ttgtatatat ctgacaaaac tggcaacatc 4907

ataaagacta ctgacttgaa gacaacctct tttatatttc tcgatttctg ggctaatgaa 4967

tttgttttca cctatctttt ccctttcaga attttattcg ggggggaaaa taactagcct 5027

agctggtcat ttctttgtaa ggtagttagc aatatttagt ctttctttgg tcaccttttt 5087

ttttttaatg tgaaaagtta agtaagaaac tttttttact gcttttatga atttctgtct 5147

tgttttgaaa ctatgatcgt tataattttg gttcccaaat aaaacattta aaaaaaataa 5207

ttaacagcaa cgtcacaaag ataatggatt gcacatagac taagaaataa acttcagaat 5267

tgtgattttt tttgtttcta atcttgatgt aaatttacac tatttataaa tacatattta 5327

ttgcctgaaa atatttgtga atggaatgct gttatttttt ccagatttac ctgccattga 5387

aattttaagg agttctgtaa tttcaaacac tactcctatt acattttcta tgtgtaaata 5447

aaactgctta gccttgtaca gaaactttta ttaaaattgt ttaatgttta aagagttttc 5507

tattgtttga gttttaaaaa gaatttatgt acagtgccca atttttgttc acttttcaaa 5567

tctgattata tatattttat atatacttat gtctgtatat ataatatata tagaaatcta 5627

gatatatgta taaagattta gaatttgaat ttttcttgtt ttaagtttca catctatggt 5687

agatttttga ggtgtctact gtaaagtatt gctcacaaaa agtatgatta tttttaaaga 5747

aatatatatg gtatgtatct tcaagaccta aaatgtcaga ctggtttatt gttaaattgc 5807

aactactgca attacagaca aaaaaaatct gccaaatgta gtataataga aaacatagtg 5867

attgaactta aaactttaaa gggagtttta atacaggctt ttcaaatcct a 5918

<210> 4

<211> 1401

<212> PRT

<213>Mouse (Mus musculus)

<400> 4

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

1 5 10 15

Ala Ala Ala Phe Gly Asp Glu Glu Lys Lys Met Ala Ala Gly Lys Ala

20 25 30

Ser Gly Glu Ser Glu Glu Ala Ser Pro Ser Leu Thr Ala Glu Glu Arg

35 40 45

Glu Ala Leu Gly Gly Leu Asp Ser Arg Leu Phe Gly Phe Val Arg Phe

50 55 60

His Glu Asp Gly Ala Arg Met Lys Ala Leu Leu Gly Lys Ala Val Arg

65 70 75 80

Cys Tyr Glu Ser Leu Ile Leu Lys Ala Glu Gly Lys Val Glu Ser Asp

85 90 95

Phe Phe Cys Gln Leu Gly His Phe Asn Leu Leu Leu Glu Asp Tyr Pro

100 105 110

Lys Ala Leu Ser Ala Tyr Gln Arg Tyr Tyr Ser Leu Gln Ser Asp Tyr

115 120 125

Trp Lys Asn Ala Ala Phe Leu Tyr Gly Leu Gly Leu Val Tyr Phe His

130 135 140

Tyr Asn Ala Phe Gln Trp Ala Ile Lys Ala Phe Gln Glu Val Leu Tyr

145 150 155 160

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

165 170 175

Leu Met Phe Lys Val Asn Thr Asp Tyr Glu Ser Ser Leu Lys His Phe

180 185 190

Gln Leu Ala Leu Val Asp Cys Asn Pro Cys Thr Leu Ser Asn Ala Glu

195 200 205

Ile Gln Phe His Ile Ala His Leu Tyr Glu Thr Gln Arg Lys Tyr His

210 215 220

Ser Ala Lys Glu Ala Tyr Glu Gln Leu Leu Gln Thr Glu Asn Leu Ser

225 230 235 240

Ala Gln Val Lys Ala Thr Ile Leu Gln Gln Leu Gly Trp Met His His

245 250 255

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

260 265 270

Gln Tyr Leu Gln Lys Ser Leu Glu Ala Asp Pro Asn Ser Gly Gln Ser

275 280 285

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

290 295 300

Ala Phe Ile Ser Tyr Arg Gln Ser Ile Asp Lys Ser Glu Ala Ser Ala

305 310 315 320

Asp Thr Trp Cys Ser Ile Gly Val Leu Tyr Gln Gln Gln Asn Gln Pro

325 330 335

Met Asp Ala Leu Gln Ala Tyr Ile Cys Ala Val Gln Leu Asp His Gly

340 345 350

His Ala Ala Ala Trp Met Asp Leu Gly Thr Leu Tyr Glu Ser Cys Asn

355 360 365

Gln Pro Gln Asp Ala Ile Lys Cys Tyr Leu Asn Ala Thr Arg Ser Lys

370 375 380

Asn Cys Ser Asn Thr Ser Gly Leu Ala Ala Arg Ile Lys Tyr Leu Gln

385 390 395 400

Ala Gln Leu Cys Asn Leu Pro Gln Gly Ser Leu Gln Asn Lys Thr Lys

405 410 415

Leu Leu Pro Ser Ile Glu Glu Ala Trp Ser Leu Pro Ile Pro Ala Glu

420 425 430

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

435 440 445

Asp Asn Trp Ser Gly Gly Asn Ala Pro Pro Pro Val Glu Gln Gln Thr

450 455 460

His Ser Trp Cys Leu Thr Pro Gln Lys Leu Gln His Leu Glu Gln Leu

465 470 475 480

Arg Ala Asn Arg Asn Asn Leu Asn Pro Ala Gln Lys Leu Met Leu Glu

485 490 495

Gln Leu Glu Ser Gln Phe Val Leu Met Gln Gln His Gln Met Arg Gln

500 505 510

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

515 520 525

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

530 535 540

Leu Pro Leu Thr Arg Met Pro Ser Val Ser Gln Pro Gly Val His Thr

545 550 555 560

Ala Cys Pro Arg Gln Thr Leu Ala Asn Gly Pro Phe Ser Ala Gly His

565 570 575

Val Pro Cys Ser Thr Ser Arg Thr Leu Gly Ser Thr Asp Thr Val Leu

580 585 590

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

595 600 605

Tyr Leu Gln Arg Asn Ala Pro Thr Leu Pro His Asn Arg Thr Asn Leu

610 615 620

Thr Ser Ser Thr Glu Glu Pro Trp Lys Asn Gln Leu Ser Asn Ser Thr

625 630 635 640

Gln Gly Leu His Lys Gly Pro Ser Ser His Leu Ala Gly Pro Asn Gly

645 650 655

Glu Arg Pro Leu Ser Ser Thr Gly Pro Ser Gln His Leu Gln Ala Ala

660 665 670

Gly Ser Gly Ile Gln Asn Gln Asn Gly His Pro Thr Leu Pro Ser Asn

675 680 685

Ser Val Thr Gln Gly Ala Ala Leu Asn His Leu Ser Ser His Thr Ala

690 695 700

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

705 710 715 720

Ser Gly Asn Thr Leu Thr Val Pro Glu Thr Ser Arg Gln Thr Gly Glu

725 730 735

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

740 745 750

Gln Val Met Ala Asp Ala Val Cys Ser Pro Ser His Gly Asp Ser Lys

755 760 765

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

770 775 780

Met Gly Lys Ala Asn Asn Asn Val Gly Pro Gly Thr Cys Asp Lys Val

785 790 795 800

Asn Asn Ile His Pro Thr Val His Thr Lys Thr Asp Asn Ser Val Ala

805 810 815

Ser Ser Pro Ser Ser Ala Ile Ser Thr Ala Thr Pro Ser Pro Lys Ser

820 825 830

Thr Glu Gln Thr Thr Thr Asn Ser Val Thr Ser Leu Asn Ser Pro His

835 840 845

Ser Gly Leu His Thr Ile Asn Gly Glu Gly Met Glu Glu Ser Gln Ser

850 855 860

Pro Ile Lys Thr Asp Leu Leu Leu Val Ser His Arg Pro Ser Pro Gln

865 870 875 880

Ile Ile Pro Ser Met Ser Val Ser Ile Tyr Pro Ser Ser Ala Glu Val

885 890 895

Leu Lys Ala Cys Arg Asn Leu Gly Lys Asn Gly Leu Ser Asn Ser Ser

900 905 910

Ile Leu Leu Asp Lys Cys Pro Pro Pro Arg Pro Pro Ser Ser Pro Tyr

915 920 925

Pro Pro Leu Pro Lys Asp Lys Leu Asn Pro Pro Thr Pro Ser Ile Tyr

930 935 940

Leu Glu Asn Lys Arg Asp Ala Phe Phe Pro Pro Leu His Gln Phe Cys

945 950 955 960

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

965 970 975

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

980 985 990

Asn Asn Glu His Met Val Glu Val Arg Thr Gln Leu Leu Gln Pro Ala

995 1000 1005

Asp Glu Asn Trp Asp Pro Thr Gly Thr Lys Lys Ile Trp His Cys

1010 1015 1020

Glu Ser Asn Arg Ser His Thr Thr Ile Ala Lys Tyr Ala Gln Tyr

1025 1030 1035

Gln Ala Ser Ser Phe Gln Glu Ser Leu Arg Glu Glu Asn Glu Lys

1040 1045 1050

Arg Ser His His Lys Asp His Ser Asp Ser Glu Ser Thr Ser Ser

1055 1060 1065

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

1070 1075 1080

Phe Gly Thr Asn Ile Asp Leu Ser Asp Asp Lys Lys Trp Lys Leu

1085 1090 1095

Gln Leu His Glu Leu Thr Lys Leu Pro Ala Phe Val Arg Val Val

1100 1105 1110

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

1115 1120 1125

Met Asn Thr Val Gln Leu Tyr Met Lys Val Pro Gly Ser Arg Thr

1130 1135 1140

Pro Gly His Gln Glu Asn Asn Asn Phe Cys Ser Val Asn Ile Asn

1145 1150 1155

Ile Gly Pro Gly Asp Cys Glu Trp Phe Val Val Pro Glu Gly Tyr

1160 1165 1170

Trp Gly Val Leu Asn Asp Phe Cys Glu Lys Asn Asn Leu Asn Phe

1175 1180 1185

Leu Met Gly Ser Trp Trp Pro Asn Leu Glu Asp Leu Tyr Glu Ala

1190 1195 1200

Asn Val Pro Val Tyr Arg Phe Ile Gln Arg Pro Gly Asp Leu Val

1205 1210 1215

Trp Ile Asn Ala Gly Thr Val His Trp Val Gln Ala Ile Gly Trp

1220 1225 1230

Cys Asn Asn Ile Ala Trp Asn Val Gly Pro Leu Thr Ala Cys Gln

1235 1240 1245

Tyr Lys Leu Ala Val Glu Arg Tyr Glu Trp Asn Lys Leu Gln Asn

1250 1255 1260

Val Lys Ser Ile Val Pro Met Val His Leu Ser Trp Asn Met Ala

1265 1270 1275

Arg Asn Ile Lys Val Ser Asp Pro Lys Leu Phe Glu Met Ile Lys

1280 1285 1290

Tyr Cys Leu Leu Arg Thr Leu Lys Gln Cys Gln Thr Leu Arg Glu

1295 1300 1305

Ala Leu Ile Ala Ala Gly Lys Glu Ile Ile Trp His Gly Arg Thr

1310 1315 1320

Lys Glu Glu Pro Ala His Tyr Cys Ser Ile Cys Glu Val Glu Val

1325 1330 1335

Phe Asp Leu Leu Phe Val Thr Asn Glu Ser Asn Ser Arg Lys Thr

1340 1345 1350

Tyr Ile Val His Cys Gln Asp Cys Ala Arg Lys Thr Ser Gly Asn

1355 1360 1365

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

1370 1375 1380

Met Gln Val Tyr Asp Gln Phe Thr Leu Ala Pro Pro Leu Pro Ser

1385 1390 1395

Ala Ser Ser

1400

<210> 5

<211> 19

<212> RNA

<213>Artificial sequence

<400> 5

gcugcuacga aucucuaau 19

<210> 6

<211> 19

<212> RNA

<213>Artificial sequence

<400> 6

auuagagauu cguagcagc 19

<210> 7

<211> 19

<212> RNA

<213>Artificial sequence

<400> 7

uucuccgaac gugucacgu 19

<210> 8

<211> 19

<212> RNA

<213>Artificial sequence

<400> 8

acgugacacg uucggagaa 19

Claims (9)

1.Kdm6a albumen or Kdm6a coded sequences, its accelerator or its inhibitor are being prepared for regulating and controlling the generation of I types interferon Medicine or kit in purposes.

2. purposes according to claim 1, it is characterised in that

The Kdm6a albumen is selected from：

(a)SEQ ID NO：2 or SEQ ID NO：Amino acid sequence shown in 4；Or

(b) with SEQ ID NO：2 or SEQ ID NO：Amino acid sequence homologous shown in 4, it, which has, suppresses virus infection correlation The albumen or polypeptide of disease and/or symptom activity；Or

(c) in the amino acid sequence of (a) or (b) by substitution, lack or add one or several amino acid and with prevention or The protein as derived from (a) or (b) or polypeptide for the treatment of virus infection relevant disease and/or symptom activity；

The Kdm6a encoding genes are selected from：

(i)SEQ ID NO：1 or SEQ ID NO：1 378-4583 bit sequences, SEQ ID NO：3 or SEQ ID NO：3 Nucleotide sequence shown in 432-4637 bit sequences；Or

(ii) molecule under strict conditions with (i) nucleotide sequence hybridization limited；Or

(iii) have in (i) or (ii) nucleotide sequence by replacing, lacking or add one or several nucleotides and encode There is the molecule of prevention or treatment virus infection relevant disease and/or symptom；

The accelerator of the Kdm6a or its coded sequence are selected from：It is the over-express vector of Kdm6a or its coded sequence, exogenous Liposome DNA, the Kdm6a albumen of the naked DNA of Kdm6a, Kdm6a or its coded sequence, Kdm6a or its coded sequence；

The inhibitor of the Kdm6a or its coded sequence are selected from for Kdm6a or its coded sequence：Antibody, siRNA, MiRNA, ASON, antagonist, blocking agent.

3. purposes according to claim 1, it is characterised in that the Kdm6a, its coded sequence or its accelerator promote I The generation of type interferon；The inhibitor suppresses the generation of I type interferon.

4. purposes according to claim 1, it is characterised in that the I types interferon is selected from：IFN-α or IFN-β.

5. purposes according to claim 1, it is characterised in that the medicine or kit are encoded comprising Kdm6a or Kdm6a Sequence or its accelerator, and by promoting the generation of I type interferon to be further used for preventing or treating related to viral infection Disease and/or its symptom；The inhibitor of the medicine or kit comprising Kdm6a or its coded sequence, and done by suppressing I types Disturb that the excessive generation of element is used to preventing or treating caused by I type interferon overexpressions tissue and organ damage and/or it is levied Shape.

6. purposes according to claim 5, it is characterised in that the virus infection relevant disease and/or symptom be selected from One or more kinds of disease and/or symptoms caused by virus infection of the following group：Virus is metainfective to be replicated；Viral infection office The cell in portion and tissue damage, the cell include exempting from the epithelial cell, the immunocyte of peripheral blood, immune organ of skin The composition cell of epidemic disease cell and solid organ, the tissue includes skin, liver, kidney, brain, lungs；Virus infection is caused Multi-organ dysfunction, the organ include skin, liver, kidney, brain, lungs.

7. purposes according to claim 6, it is characterised in that the virus infection is the one or two by being selected from the group Caused by above virus：Human papilloma virus, hepatitis B, Epstein-Barr virus, epidemic hemorrhagic fever virus, people's acquired immunity lack Fall into virus, SARS virus, influenza virus.

8. purposes according to claim 1, it is characterised in that the medication of the medicine is selected from：

Give Kdm6a coded sequences：Including direct naked DNA injection method, liposome DNA direct injections, gold coating DNA bases Target DNA method is carried because rifle blast technique, breeding unsoundness bacterium carry DNA method, replication defective adenoviral；

Give Kdm6a albumen：Including drug administration by injection, direct injection Kdm6a albumen or with liposome embedded Kdm6a3 albumen, nose Chamber administration, pulmonary administration, oral administration, cutaneous penetration.

9. a kind of be used to regulate and control the pharmaceutical composition that I types interferon is produced, it is characterised in that includes：

(A) the Kdm6a albumen or Kdm6a coded sequences of therapeutically effective amount, its accelerator or its inhibitor；

(B) prevent or treat virus infection relevant disease and/or other active materials of symptom activity；And

(C) acceptable carrier or excipient pharmaceutically or in immunology.