CN109022487A - A kind of genophore for expressing p65 and its gene therapy medicament for treating retinal ganglion cells degeneration - Google Patents

Abstract

A kind of gene therapy medicament the present invention relates to genophore for expressing p65 and its for treating retinal ganglion cells degeneration, (constitutively active) mankind p65 (p65 (CA)) of the genophore comprising activated form, promoter/enhancer, the β-globin introne for increasing gene expression, human growth hormone poly (A) tail sequence etc..Application of the composition the invention further relates to said gene carrier or comprising said gene carrier in the drug of preparation treatment retinal ganglion cells degeneration.Its advantage is shown: gene therapy vector of the invention can be used to treat or prevent the retinal ganglial cells lesion as caused by glaucoma etc..

Description

It is a kind of express p65 genophore and its for treat retinal ganglial cells become The gene therapy medicament of property

Technical field

The present invention relates to gene engineering technology fields, specifically, be it is a kind of express p65 genophore and its for controlling Treat the gene therapy medicament of retinal ganglion cells degeneration.

Background technique

Retinal ganglial cells (Retinal ganglion cells, RGCs) denaturation is found in a variety of acute and chronic views Film lesion, such as glaucoma, diabetic retinopathy, optic neuritis, the death of the RGC as caused by these diseases and axonal injury Finally the visual function of progressive is caused to be lost.It there is no effective intervening measure for RGC death at present, at present majority treatment means For promoting the effect of the survival for being damaged RGC and axon regeneration still extremely limited.Therefore, it finds new protection RGC and then saves The therapeutic strategy of patient's vision is particularly important.

NF κ B signal access for adjust inflammatory response, the cell activities such as cell survival and cell Proliferation to close weight It wants.Some researches show that NF κ B access has the function of inhibiting neuronal death and adjusts neure growth.The subunit p50 of NF κ B The hippocampus of mice cone neurone of missing is decreased obviously the tolerance of neurotoxin；P65 is another subunit of NF κ B, water Flat reduction accelerates the death of dynamoneure；On the other hand, the overexpression of p65 can significantly inhibit low potassium induction The apoptosis of cerebellar granule neuron；The neuroprotection of NF κ B access is also confirmed in sympathetic neuron.In addition, a system Column nerve growth factor (nerve growth factor, NGF) such as BDNF, NT-3, NT-4/5 etc. can be active cell core NF κ B turns Record factor complex, NF κ B by adjust cytokine profiles, inflammatory mediator, the protection of the mediate neuronals such as cell adhesion molecule and Growth promoting function.The gene for having studies have shown that neuron targeting knockout NF κ B to rely on can accelerate the nerve degeneration of forehead leaf texture.

RGC belongs to a part of central nervous system, is responsible for exporting the visual signal captured by retina to brain.NF κ B signal access is in silent status in normal RGC, and after axonal injury major part RGC can not independently activate NF κ B access and Final apoptosis has research to confirm that activation NF κ B access can promote RGC and survive after axonal injury, NF κ B access is prompted to exist RGC It survives after axonal injury very crucial.Therefore, in various optic nerve degeneration diseases, enhancing NF κ B signal access in RGC may It is that RGC is prevented to be denaturalized, promotes effective remedy measures of RGC survival and axon regeneration.

Therefore, present invention combination AAV technology develops a kind of special target RGC and expresses continuous activation The gene therapy vector of the p65 (CA) of (constitutively active).

Summary of the invention

The first purpose of this invention is aiming at the shortcomings in the prior art, to provide a kind of genophore.

Second object of the present invention is to provide a kind of pharmaceutical composition.

Third object of the present invention is to provide a kind of application of genophore.

Fourth object of the present invention is to provide a kind of method for treating retinal ganglion cells degeneration.

To realize above-mentioned first purpose, the technical solution adopted by the present invention is that: a kind of adeno-associated virus load for expressing p65 Body, the base that the gland relevant viral vector contains enhancer, specificity or non-specific promoter, expresses p65 (CA) albumen Because of sequence, the sequence of p65 (CA) albumen is as shown in SEQ ID NO:2.

Preferably, the gene order for expressing p65 (CA) albumen, which is selected from, has at least 96% with SEQ ID NO:1,97%, 98%, the nucleotide sequence of 99%, 100% homology.

It is highly preferred that the gene order of the p65 (CA) contains SEQ ID NO:1.

Most preferably, the nucleotide sequence of the p65 (CA) albumen is selected from SEQ ID NO:1.

In any of the above-described preference, enhancers/promoters CMV is at least 95% homologous selected from having with SEQ ID NO:4 The nucleotide sequence of property.It is highly preferred that the CMV be selected from have at least 96% with SEQ ID NO:4,97%, 98%, 99%, The nucleotide sequence of 100% homology.Most preferably, the nucleotide sequence of the cmv enhancer is selected from SEQ ID NO:4.

In any of the above-described preference, adeno-associated virus can be selected from AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV2-AAV3, AAVrh.10, AAVhu.14, AAV3a/3b, AAVrh32.33, AAVHSC15, AAV-HSC17, AAVhu.37, AAVrh.8, CHt-P6, AAV2.5, AAV6.2, AAV2i8, AAV-HSC15/17, AAVM41, AAV9.45, AAV6 (Y445F/Y731F), AAV2.5T, AAV-HAE1/2, AAV clone 32/83, AAVShH10, AAV2 (Y- > F), AAV8 (Y733F), AAV2.15, AAV2.4, AAVM41, AAVr3.45AAV2 or AAV5.Most preferably, gland phase It closes virus and is selected from AAV2.

In any of the above-described preference, adeno-associated virus is single-stranded AAV.

In any of the above-described preference, the chimeric interon that the gland relevant viral vector also shortens containing one section, institute The chimeric intron sequence stated is selected from the nucleotide sequence for having at least 90% homology with SEQ ID NO:5.

In any of the above-described preference, the gland relevant viral vector contains cmv enhancer/promoter, p65 (CA) Gene order, β-globin introne, hGHpA, L-ITR, R-ITR, Ampicillin, f1ori.

In any of the above-described preference, the gene order of p65 (CA) albumen is expressed as shown in SEQ ID NO:1, L-ITR sequence Column are as shown in SEQ ID NO:3, and shown in cmv enhancer/promoter sequence SEQ ID NO:4, β-globin intron sequences are such as Shown in SEQ ID NO:5, hGHpA sequence as shown in SEQ ID NO:7, R-ITR sequence as shown in SEQ ID NO:8, Ampicillin sequence is as shown in SEQ ID NO:9, and f1ori sequence is as shown in SEQ ID NO:11.

In any of the above-described preference, the gland relevant viral vector sequence contains SEQ ID NO:10.

To realize above-mentioned second purpose, the technical solution adopted by the present invention is that: it is a kind of to treat retinal ganglial cells The pharmaceutical composition of denaturation, containing above-mentioned gland relevant viral vector and pharmaceutically acceptable carrier, the pharmaceutical composition Express p65 (CA) albumen.

To realize above-mentioned third purpose, the technical solution adopted by the present invention is that: it is prepared by above-mentioned gland relevant viral vector Treat the application in the drug of retinal ganglion cells degeneration.

To realize above-mentioned 4th purpose, the technical solution adopted by the present invention is that: a kind of gene therapy ganglia retinae The method of cell degeneration, by gland relevant viral vector intraocular injection, the p65 albumen of continuous expression activation.Preferably, the eye Interior injection refers to subretinal space injection or intravitreal.

The invention has the advantages that:

Gene therapy vector of the invention is for treating or preventing the disease of the retinal ganglial cells as caused by glaucoma etc. Become.

Detailed description of the invention

Attached drawing 1 is the Plasmid pattern figure of AAV_p65 (CA) carrier.

Attached drawing 2 is the plasmid characteristic pattern of AAV_p65 (CA).

Attached drawing 3 is C57BL/6J mouse intravitreal AAV_p65 (CA) inner nuclear layer retina after two weeks, immunofluorescence dye Chromatic graph.

Attached drawing 4 is C57BL/6J mouse intravitreal AAV_p65 (CA) or AAV-GFP, and is being injected at the moment simultaneously Side's injection bead particulates cause glaucoma, and the inner nuclear layer retina after 2 weeks, 4 weeks, 8 weeks is illustrated as its immunofluorescence dyeing Figure.

Attached drawing 5 is C57BL/6J mouse intravitreal AAV_p65 (CA) or AAV-GFP and is injecting side at the moment simultaneously Injection bead particulates cause glaucoma, and it is laggard to be illustrated as its immunofluorescence dyeing for the inner nuclear layer retina after 2 weeks, 4 weeks, 8 weeks Row RGC is counted and survival rate analysis result figure.

Specific embodiment

The invention will be further elucidated with reference to specific embodiments.It should be understood that these embodiments are merely to illustrate this hair It is bright rather than limit the scope of the invention.In addition, it should also be understood that, after having read the content of the invention recorded, art technology Personnel can make various changes or modifications the present invention, and such equivalent forms equally fall within the application the appended claims and limited Fixed range.

The present inventor after study, provides a kind of gland relevant viral vector and its becomes for treating retinal ganglial cells Related drugs, the pharmaceutical composition etc. of property.

Pharmaceutical composition and medicine box

For the ease of clinical application, pharmaceutical composition of the invention may be embodied in injection delivery device (such as pumping needle) In, in the injection delivery device, it may include the pharmaceutical composition of single administration amount.The injection administration Device can be contained in medicine box, to facilitate storage, use.It needs to place the small container equipped with drug suspension when transport In dry ice.It should usually be stored in -80 DEG C of refrigerators.

In medicine box or kit of the present invention, it may also include operation instructions, so that those skilled in the art press It is used according to correct mode.

As used herein, the ingredient of " pharmaceutically acceptable " is suitable for people and/or mammal and without excessively bad Side reaction (such as toxicity), i.e., with the substance of reasonable benefit/risk ratio.Term " pharmaceutically acceptable carrier ", which refers to, to be used for The carrier of Therapeutic Administration, including various excipient and diluent.This, which belongs to, refers to medicament carriers some in this way: themselves is not It is necessary active constituent, and does not have excessive toxicity after applying.

Suitable pharmaceutically acceptable carrier is well known to those of ordinary skill in the art.In Remington ' It can find in sPharmaceutical Sciences (Mack Pub.Co., N.J.1991) about pharmaceutically acceptable carrier Absolutely prove.Pharmaceutically acceptable carrier can contain liquid in the composition, such as water, BBS (Balanced SaltSolution) phosphate buffer, ringer solution, physiological saline, balanced salt solution, glycerol or sorbierite etc..In addition, There is likely to be complementary substance in these carriers, as lubricant, glidant, wetting agent or emulsifier, pH buffer substance and Stabilizer etc..

Gene therapy vector

Gene therapy vector in the present invention is virus expression carrier, and according to the present invention, virus expression carrier is gland correlation Viral (AAV) carrier is such as formed selected from chimeric AAV derived from serotypes A AV1,2,3,4,5,6,7,8,9 and 10 or its Group in AAV carrier such as AAV2-AAV3, AAVrh.10, AAVhu.14, AAV3a/3b, AAVrh32.33, AAVHSC15, AAV-HSC17, AAVhu.37, AAVrh.8, CHt-P6, AAV2.5, AAV6.2, AAV2i8, AAV-HSC15/17, AAVM41, AAV9.45, AAV6 (Y445F/Y731F), AAV2.5T, AAV-HAE1/2, AAV clone 32/83, AAVShH10, AAV2 (Y- > F), AAV8 (Y733F), AAV2.15, AAV2.4, AAVM41, AAVr3.45AAV2 or AAV5, this can be better adapted for feeling emerging Efficient transduction is carried out in the tissue of interest.Transfection when, AAV only cause in host slight immune response (if there is Words).In a preferred embodiment of the invention, gene therapy vector is AAV serotype 2 or 5 carriers.In further preferred reality It applies in mode, gene therapy vector is 2 carrier of AAV.

AAV carrier of the invention is single-stranded AAV, can produce recombinant viral vector according to standard technique.For example, recombination gland phase Closing viral vectors can propagate in 293 cell of people (it provides trans- E1A and E1B characteristic), to reach in 10^7~10^13 disease Titre within the scope of malicious particle/mL.Before applying in vivo, viral vectors can pass through gel filtration method (such as agarose column) Desalination is carried out, and is purified by subsequent filtering.Purifying reduces potential illeffects in the main body of drug administration carrier.It is given The virus of medicine is substantially free of wild-type virus and replication competent type virus.Suitable method, such as dodecyl sulphate can be passed through Sodium-polyacrylamide gel electrophoresis (SDS-PAGE), then carries out silver staining, to prove the purity of virus.

The suitable dose of AAV for people is in the range of about 1 × 10^10~1 × 10^14 virion.

Gene order

It is p65 (CA) sequence after optimization shown in SEQ ID NO:1,

It is original people p65 (CA) amino acid sequence shown in SEQ ID NO:2,

It is Left ITR sequence shown in SEQ ID NO:3,

It is CMV sequence shown in SEQ ID NO:4,

It is human β-globin intron sequence shown in SEQ ID NO:5,

It is MCS sequence shown in SEQ ID NO:6,

It is hGHpA sequence shown in SEQ ID NO:7,

It is Right ITR sequence shown in SEQ ID NO:8,

It is Ampicillin Resistance sequence shown in SEQ ID NO:9,

It is full nucleotide sequence shown in SEQ ID NO:10,

It is f1ori sequence shown in SEQ ID NO:11,

It is recombination sequence shown in SEQ ID NO:12.

Gene therapy vector is intraocular injection administration, can be administered using subretinal space or intravitreal.Under In conjunction with specific embodiments, the present invention is further explained in face.It should be understood that these embodiments are merely to illustrate the present invention rather than limit The scope of the present invention processed.

In the following examples, the experimental methods for specific conditions are not specified, usually according to normal condition such as J. Pehanorm Brooker It writes, Molecular Cloning:A Laboratory guide, the third edition, Science Press, condition described in 2002, or according to proposed by manufacturer Condition.

Experimental animal

Normal C57BL/6J mouse is purchased from Shanghai Slac Experimental Animal Co., Ltd..Periodicity of illumination 12h illumination- 12h is dark, is freely eaten, free water, " the experiment that all zooscopies are issued in strict accordance with State Scientific and Technological Commission The care of animal regulations " it carries out.

Main agents, consumptive material

Physiological saline (Zhejiang Tianrui Pharmaceutical Co., Ltd.), injection needles head (U.S. Becton Dickinson And Company company), HA (Roche), TUJ1 (Abcam), secondary antibody goat anti-rabbit igg (JacksonImmuno), blood of goats (Sigma) clearly, Triton X-100 (Triton X-100) (Sigma), paraformaldehyde (Sigma).

Key instrument

Ophthalmology tests surgical operation microscope (Japanese Nikon company)；Ophthalmology microinstrument (the limited public affairs of Suzhou Ming Ren medical instrument Department)；Microsyringe (Hamilton company of the U.S.)；Laser Scanning Confocal Microscope (Leca company).

The building of mouse glaucoma model

The injection that is prepared in advance capillary needle tubing, needle tubing top is oval, and interior to angle there are 20 degree, and prepares The good bead particulates suspension (1.6*10^beads/ul) for having removed surface epoxy group.It chooses 6-8 weeks male mice and carries out abdominal cavity Injecting anesthetic, side are placed on station, and modeling eye needs level upward and need to expand pupil in advance.Operation consent is magnetic by preprepared Microparticulate suspensions blow and beat mixing again, and draw 1.5ul magnetic particle suspension in capillary needle tubing rapidly.Capillary needle tubing and corneal limbus It is therewith in fix eyeball with plastic cement microforceps at 90 degree in 45 degree of angles, and in the other side.It is careful to be pierced into capillary needle tubing, it needs to guarantee Needle tubing does not enter front depths, and forbids touching iris and crystal in the process.Injection point at side angle Gong Yuan place a magnetic pole, Slowly injection 1.5ul magnetic particle suspension, injection process need to guarantee to complete in 15 to 30 seconds afterwards, need to guarantee magnetic in injection process Pole position is constant.Capillary needle tubing is removed after the completion of injection rapidly, rear position of magnetic pole remains unchanged, and maintenance 30 to 60 seconds to guarantee There is particle to be not attached in corneal limbus to survey.Canthus Gong Yuan is that magnetic particle is dispersed in angle around the mobile magnetic pole of eyeball later Film, iris angle simultaneously form magnetic particle ring.Mouse needs side to be placed on hot blanket to recover after modeling, and resuscitation process needs to keep Modeling eye is upward.

The preparation of magnetic micro-beads aaerosol solution: because magnetic micro-beads surface is covered with epoxy group, should first remove before use, i.e., will 1ml magnetic micro-beads are mixed into rapidly the 10*Tris of the 0.02M sodium hydroxide (NaOH, MW 39.997g/mol) of the fresh configuration of 50ml In the mixed liquor of buffer, shaken at room temperature is mixed 24 hours, is collected bead particulates in test tube bottom using magnetic force.Test tube is laid flat To ensure that all particles are all adsorbed by magnetic force.It mixes 4 hours at room temperature.It is carefully gone out supernatant liquid with pipettor, carefully Magnetic bead group is mixed into 50ml 10X Tris solution, mixes and makes all particle suspensions.Bead particulates are in sterile equilibrium liquid Concentration and suspension: experiment is used ultrapure water oscillation cleaning particle 3 times.In super-clean bench, 500mlBBS piping and druming is added and mixes 3 times.Add Enter 250mlBBS piping and druming to mix, makes particle resuspension.Ensure that solution is uniform as far as possible, and quickly by 25ul suspension move into 0.5ml without In tube.Obtained Nanoparticle Solution ultimate density is 1.6*10^6/ul.4 DEG C of preservations.

Intravitreal

Using glass system capillary needle, locates inserting needle rearward in view film edge, corneosclera boundary, infused using micro syringe It penetrates, when injection should be noted needle angle and depth, pay attention to avoiding damage to crystal or other eye inner tissues.

Tissue treatment

Animal first via PBS carry out total body perfusion, after continue with 4% paraformaldehyde (PFA) perfusion, rate of flooding 5mL/ Minute.Eyeball and optic nerve impregnate about 2 hours after taking out in 4%PFA solution.It need to be in 0.1M PBS before tissue staining tile 4 DEG C of storages；Before tissue freezing section need in 30% sucrose solution soaked overnight.

Embodiment 1: it the building of adenovirus vector and isolates and purifies

The building of plasmid AAV_p65 (CA) is as shown in Figure 1 and Figure 2, and main element includes cmv enhancer/promoter and p65 (CA) sequence.The expression of transgene can be enhanced in cmv enhancer.After target gene followed by hGHpA (SEQ ID NO: 7), expression cassette two sides are inverted terminal repeat (TR), i.e., viral vectors includes L-ITR (SEQ ID NO:3), R-ITR (SEQ ID NO:8), viral vectors further include Ampicillin (SEQ ID NO:9) and f1ori (SEQ ID NO:11).

Viral vectors is obtained by plasmid co-transfection method.Containing AAV2 coat protein gene and it will can help AAV It is related to preliminarily form recombination gland for helper plasmid and AAV_p65 (CA) plasmid and HEK 293T cell cotransfection of the gene of duplication Viral vectors.After iodine gram butanol preliminary purification, by the fast protein liquid using 5ml-Hitrp Q Ago-Gel as filler Chromatography is further purified by ion-exchange chromatography, the use of instrument is Pharmacia AKTA FPLC system (Amersham Biosciences,Piscataway,NJ).PH8.0, the NaCl of 215mM is used to elute agarose Gel column later, The recombinant viral vector of peak value is collected.The liquid of collection by inspissator (100K concentrater, Millipore after), recombinant viral vector is concentrated with the polysorbas20 elution inspissator containing 0.014%.DNase I is used again DNA other than virion is digested, and determines the titre of virus by the method for real-time fluorescence quantitative PCR.Finally use silver nitrate Dyeing-SDS polyacrylamide gel electrophoresis method ensures that vector particles are contaminated and are free of endotoxin, and dispenses subzero 80 Degree storage.

The characteristic spectrum of viral vectors AAV_p65 (CA) is as shown in Figure 1, the start-stop site of each element is as shown in Figure 2.

Embodiment 2: adenovirus infection detection

Experimental method:

Tissue treatment: mouse intravitreal HA label AAV_p65 (CA) after two weeks, according to aforementioned tissues processing method Carry out the pre-treatment of eyeball, clamp eyeball bottom using microforceps afterwards and be lifted up, with scissors cut peribulbar conjunctiva and Musculature keeps eyeball tissue complete, cuts off cornea, iris and crystalline lens and be made into eyecup, and separate retina and choroid Complex.

Immunofluorescence dyeing: choosing retina, and 1h is closed in 20% goat serum solution, and rear primary antibody is incubated overnight, and resists Body is respectively TUJ1, HA；10min, rear secondary antibody are incubated for 1h three times, every time for PBS cleaning；PBS cleaning is sealed ten minutes later three times, every time Piece.Change and take pictures using confocal microscopy retinal structure.

As a result: mouse injection of AAV _ p65 (CA) after two weeks, carries out retina immunofluorescence dyeing tile, it is seen that deposit on eyeground In the AAVp65 (CA) of a large amount of HA label, and it is overlapped (TUJ1 immunofluorescence dyeing) with RGC, shows the infection of AAV_p65 (CA) Excellent effect (Fig. 3).

The detection of embodiment 3:RGC survival rate

Experimental method: 8 week old mouse unilateral side camera vitrea bulbi injection of AAV _ p65 (CA) (experimental group), AAV-GFP (control Group), while the injection of anterior chamber's bead particulates is carried out in injection eye, eyeball and view were handled according to preceding method respectively at 2 weeks, 4 weeks, 8 weeks Nethike embrane carries out immunofluorescence dyeing.Change and take pictures using confocal microscopy retinal structure.Each eyeball selects at random It takes 5 visuals field to count RGC (scale bar: 20um), and calculates RGC survival rate.

As a result: quantitative analysis proves RGC gradually apoptosis in 2W, 4W, 8W after anterior chamber's bead injection, and quantity reduce it is significant (experimental group)；In contrast, injection of AAV _ p65 (CA) mouse RGC apoptosis is not significant, display AAV_p65 (CA), which has RGC, to be protected Shield effect.

The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art Member, under the premise of not departing from the method for the present invention, can also make several improvement and supplement, these are improved and supplement also should be regarded as Protection scope of the present invention.

SEQUENCE LISTING

<110>No.1 People's Hospital Shanghai City

<120>a kind of genophore for expressing p65 and its gene therapy medicine for treating retinal ganglion cells degeneration

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cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgt caatagggac 180

tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca 240

agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg 300

gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt 360

agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg 420

gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg 480

caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg 540

ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag 600

atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc 660

agc 663

<210> 5

<211> 493

<212> DNA

<213>artificial sequence

<400> 5

gattcgaatc ccggccggga acggtgcatt ggaacgcgga ttccccgtgc caagagtgac 60

gtaagtaccg cctatagagt ctataggccc acaaaaaatg ctttcttctt ttaatatact 120

tttttgttta tcttatttct aatactttcc ctaatctctt tctttcaggg caataatgat 180

acaatgtatc atgcctcttt gcaccattct aaagaataac agtgataatt tctgggttaa 240

ggcaatagca atatttctgc atataaatat ttctgcatat aaattgtaac tgatgtaaga 300

ggtttcatat tgctaatagc agctacaatc cagctaccat tctgctttta ttttatggtt 360

gggataaggc tggattattc tgagtccaag ctaggccctt ttgctaatca tgttcatacc 420

tcttatcttc ctcccacagc tcctgggcaa cgtgctggtc tgtgtgctgg cccatcactt 480

tggcaaagaa ttg 493

<210> 6

<211> 76

<212> DNA

<213>artificial sequence

<400> 6

atcgattgaa ttccccgggg atcctctaga gtcgacctgc agaagcttgc ctcgagcagc 60

gctgctcgag agatct 76

<210> 7

<211> 479

<212> DNA

<213>artificial sequence

<400> 7

acgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc 60

cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt 120

ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga 180

caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt 240

ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt 300

tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac 360

ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg atctacccac 420

cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtcctt 479

<210> 8

<211> 141

<212> DNA

<213>artificial sequence

<400> 8

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc 120

gagcgcgcag ctgcctgcag g 141

<210> 9

<211> 858

<212> DNA

<213>artificial sequence

<400> 9

gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt 60

ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg 120

agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga 180

agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg 240

tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt 300

tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg 360

cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg 420

aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga 480

tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc 540

tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc 600

ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc 660

ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg 720

cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac 780

gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc 840

actgattaag cattggta 858

<210> 10

<211> 4654

<212> DNA

<213>artificial sequence

<400> 10

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 120

actccatcac taggggttcc tgcggccgca cgcgtggaat tcgctagtta ttaatagtaa 180

tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg 240

gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg 300

tatgttccca tagtaacgtc aatagggact ttccattgac gtcaatgggt ggagtattta 360

cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt 420

gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac 480

tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt 540

tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac 600

cccattgacg tcaatgggag tttgttttgc accaaaatca acgggacttt ccaaaatgtc 660

gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata 720

taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg 780

acctccatag aagacaccgg gaccgatcca gcctccgcgg attcgaatcc cggccgggaa 840

cggtgcattg gaacgcggat tccccgtgcc aagagtgacg taagtaccgc ctatagagtc 900

tataggccca caaaaaatgc tttcttcttt taatatactt ttttgtttat cttatttcta 960

atactttccc taatctcttt ctttcagggc aataatgata caatgtatca tgcctctttg 1020

caccattcta aagaataaca gtgataattt ctgggttaag gcaatagcaa tatttctgca 1080

tataaatatt tctgcatata aattgtaact gatgtaagag gtttcatatt gctaatagca 1140

gctacaatcc agctaccatt ctgcttttat tttatggttg ggataaggct ggattattct 1200

gagtccaagc taggcccttt tgctaatcat gttcatacct cttatcttcc tcccacagct 1260

cctgggcaac gtgctggtct gtgtgctggc ccatcacttt ggcaaagaat tgggattcga 1320

acatcgattg aattccccgg ggatcctcta gagtcgacct gcagaagctt gcctcgagca 1380

gcgctgctcg agagatctac gggtggcatc cctgtgaccc ctccccagtg cctctcctgg 1440

ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca 1500

ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca 1560

aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga 1620

gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc 1680

tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt 1740

tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat 1800

ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg 1860

ctcccttccc tgtccttctg attttgtagg taaccacgtg cggaccgagc ggccgcagga 1920

acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg 1980

gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc 2040

gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat 2100

ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg 2160

gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct 2220

cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta 2280

aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa 2340

cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct 2400

ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc 2460

aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg 2520

ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt 2580

acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc 2640

cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct 2700

tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca 2760

ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg 2820

ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct 2880

atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga 2940

taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc 3000

cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg 3060

aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc 3120

aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact 3180

tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc 3240

ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag 3300

catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat 3360

aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt 3420

ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa 3480

gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc 3540

aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg 3600

gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt 3660

gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca 3720

gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat 3780

gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca 3840

gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg 3900

atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg 3960

ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt 4020

ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg 4080

ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata 4140

ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca 4200

ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag 4260

tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc 4320

tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga 4380

tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg 4440

tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac 4500

gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg 4560

tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg 4620

ttcctggcct tttgctggcc ttttgctcac atgt 4654

<210> 11

<211> 306

<212> DNA

<213>artificial sequence

<400> 11

cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa 60

gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc 120

aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt 180

cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca 240

acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc 300

tattgg 306

<210> 12

<211> 6271

<212> DNA

<213>artificial sequence

<400> 12

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 120

actccatcac taggggttcc tgcggccgca cgcgtggaat tcgctagtta ttaatagtaa 180

tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg 240

gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg 300

tatgttccca tagtaacgtc aatagggact ttccattgac gtcaatgggt ggagtattta 360

cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt 420

gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac 480

tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt 540

tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac 600

cccattgacg tcaatgggag tttgttttgc accaaaatca acgggacttt ccaaaatgtc 660

gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata 720

taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg 780

acctccatag aagacaccgg gaccgatcca gcctccgcgg attcgaatcc cggccgggaa 840

cggtgcattg gaacgcggat tccccgtgcc aagagtgacg taagtaccgc ctatagagtc 900

tataggccca caaaaaatgc tttcttcttt taatatactt ttttgtttat cttatttcta 960

atactttccc taatctcttt ctttcagggc aataatgata caatgtatca tgcctctttg 1020

caccattcta aagaataaca gtgataattt ctgggttaag gcaatagcaa tatttctgca 1080

tataaatatt tctgcatata aattgtaact gatgtaagag gtttcatatt gctaatagca 1140

gctacaatcc agctaccatt ctgcttttat tttatggttg ggataaggct ggattattct 1200

gagtccaagc taggcccttt tgctaatcat gttcatacct cttatcttcc tcccacagct 1260

cctgggcaac gtgctggtct gtgtgctggc ccatcacttt ggcaaagaat tgggattcga 1320

acatcgattg aattccacca tggacgaact gttccccctc atcttcccgg cagagccagc 1380

ccaggcctct ggcccctatg tggagatcat tgagcagccc aagcagcggg gcatgcgctt 1440

ccgctacaag tgcgaggggc gctccgcggg cagcatccca ggcgagagga gcacagatac 1500

caccaagacc caccccacca tcaagatcaa tggctacaca ggaccaggga cagtgcgcat 1560

ctccctggtc accaaggacc ctcctcaccg gcctcacccc cacgagcttg taggaaagga 1620

ctgccgggat ggcttctatg aggctgagct ctgcccggac cgctgcatcc acagtttcca 1680

gaacctggga atccagtgtg tgaagaagcg ggacctggag caggctatca gtcagcgcat 1740

ccagaccaac aacaacccct tccaagttcc tatagaagag cagcgtgggg actacgacct 1800

gaatgctgtg cggctctgct tccaggtgac agtgcgggac ccatcaggca ggcccctccg 1860

cctgccgcct gtcctttctc atcccatctt tgacaatcgt gcccccaaca ctgccgagct 1920

caagatctgc cgagtgaacc gaaactctgg cagctgcctc ggtggggatg agatcttcct 1980

actgtgtgac aaggtgcaga aagaggacat tgaggtgtat ttcacgggac caggctggga 2040

ggcccgaggc tccttttcgc aagctgatgt gcaccgacaa gtggccattg tgttccggac 2100

ccctccctac gcagacccca gcctgcaggc tcctgtgcgt gtctccatgc agctgcggcg 2160

gcctgacgac cgggagctca gtgagcccat ggaattccag tacctgccag atacagacga 2220

tcgtcaccgg attgaggaga aacgtaaaag gacatatgag accttcaaga gcatcatgaa 2280

gaagagtcct ttcagcggac ccaccgaccc ccggcctcca cctcgacgca ttgctgtgcc 2340

ttcccgcagc tcagcttctg tccccaagcc agcaccccag ccctatccct ttacgtcatc 2400

cctgagcacc atcaactatg atgagtttcc caccatggtg tttccttctg ggcagatcag 2460

ccaggcctcg gccttggccc cggcccctcc ccaagtcctg ccccaggctc cagcccctgc 2520

ccctgctcca gccatggtat cagctctggc ccaggcccca gcccctgtcc cagtcctagc 2580

cccaggccct cctcaggctg tggccccacc tgcccccaag cccacccagg ctggggaagg 2640

aacgctgtca gaggccctgc tgcagctgca gtttgatgat gaagacctgg gggccttgct 2700

tggcaacagc acagacccag ctgtgttcac agacctggca tccgtcgaca actccgagtt 2760

tcagcagctg ctgaaccagg gcatacctgt ggccccccac acaactgagc ccatgctgat 2820

ggagtaccct gaggctataa ctcgcctagt gacaggggcc cagaggcccc ccgacccagc 2880

tcctgctcca ctgggggccc cggggctccc caatggcctc ctttcaggag atgaagactt 2940

ctccgccatt gcggacatgg acttctcagc cctgctgagt cagatcagct cctaacagcg 3000

ctgctcgaga gatctacggg tggcatccct gtgacccctc cccagtgcct ctcctggccc 3060

tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt 3120

tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg 3180

ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg 3240

cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc 3300

ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc taatttttgt 3360

ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac tcctaatctc 3420

aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga accactgctc 3480

ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc 3540

cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg 3600

accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg 3660

cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc 3720

acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg 3780

ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct 3840

ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat 3900

cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt 3960

gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg 4020

acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac 4080

cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta 4140

aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca 4200

attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga 4260

cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 4320

agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 4380

aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata 4440

ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt 4500

tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa 4560

atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt 4620

attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa 4680

gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac 4740

agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt 4800

aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt 4860

cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat 4920

cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac 4980

actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg 5040

cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc 5100

ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa 5160

ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag 5220

gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct 5280

gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat 5340

ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa 5400

cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac 5460

caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc 5520

taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc 5580

cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg 5640

cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg 5700

gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca 5760

aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg 5820

cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg 5880

tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga 5940

acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac 6000

ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat 6060

ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc 6120

tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga 6180

tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc 6240

ctggcctttt gctggccttt tgctcacatg t 6271

Claims (10)

1. it is a kind of express p65 gland relevant viral vector, which is characterized in that the gland relevant viral vector contain enhancer, Specificity or non-specific promoter, the gene order for expressing p65 (CA) albumen, the sequence such as SEQ ID of p65 (CA) albumen Shown in NO:2.

2. gland relevant viral vector according to claim 1, which is characterized in that the gene order of expression p65 (CA) albumen As shown in SEQ ID NO:1.

3. gland relevant viral vector according to claim 1, which is characterized in that the enhancers/promoters are CMV.

4. gland relevant viral vector according to claim 1, which is characterized in that the adeno-associated virus is AAV2.

5. gland relevant viral vector according to claim 1, which is characterized in that the adeno-associated virus is single-stranded AAV.

6. gland relevant viral vector according to claim 1, which is characterized in that the gland relevant viral vector also contains The chimeric interon of one section of shortening, the chimeric intron sequence, which is selected from, has at least 90% homology with SEQ ID NO:5 Nucleotide sequence.

7. gland relevant viral vector according to claim 1, which is characterized in that the gland relevant viral vector contains Cmv enhancer/promoter, the gene order of p65 (CA), β-globin introne, hGHpA, L-ITR, R-ITR, Ampicillin、f1ori。

8. gland relevant viral vector according to claim 7, which is characterized in that the gene order of expression p65 (CA) albumen As shown in SEQ ID NO:1, L-ITR sequence is as shown in SEQ ID NO:3, cmv enhancer/promoter sequence SEQ ID NO:4 Shown, β-globin intron sequences are as shown in SEQ ID NO:5, and hGHpA sequence is as shown in SEQ ID NO:7, R-ITR sequence As shown in SEQ ID NO:8, Ampicillin sequence is as shown in SEQ ID NO:9, f1ori sequence such as SEQ ID NO:11 institute Show.

9. a kind of pharmaceutical composition for treating retinal ganglion cells degeneration, which is characterized in that the pharmaceutical composition contains There are gland relevant viral vector and pharmaceutically acceptable carrier a method as claimed in any one of claims 1-8, the pharmaceutical composition Express p65 (CA) albumen.

10. -8 any gland relevant viral vectors treat retinal ganglion cells degeneration in preparation according to claim 1 Drug in application.