CN106866797A - J subgroup avian leucosis virus specific antigen epitope, fusion protein, specific antibody and its application - Google Patents

J subgroup avian leucosis virus specific antigen epitope, fusion protein, specific antibody and its application Download PDF

Info

Publication number
CN106866797A
CN106866797A CN201710183203.2A CN201710183203A CN106866797A CN 106866797 A CN106866797 A CN 106866797A CN 201710183203 A CN201710183203 A CN 201710183203A CN 106866797 A CN106866797 A CN 106866797A
Authority
CN
China
Prior art keywords
subgroup
avian leucosis
subgroup avian
fusion protein
specific
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN201710183203.2A
Other languages
Chinese (zh)
Inventor
王永强
曹红
李晓齐
郑世军
陈福勇
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
China Agricultural University
Original Assignee
China Agricultural University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by China Agricultural University filed Critical China Agricultural University
Priority to CN201710183203.2A priority Critical patent/CN106866797A/en
Publication of CN106866797A publication Critical patent/CN106866797A/en
Pending legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56983—Viruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/11011—Alpharetrovirus, e.g. avian leucosis virus
    • C12N2740/11022—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/11011—Alpharetrovirus, e.g. avian leucosis virus
    • C12N2740/11034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2469/00—Immunoassays for the detection of microorganisms
    • G01N2469/20—Detection of antibodies in sample from host which are directed against antigens from microorganisms

Abstract

The invention belongs to Animal diseases detection technique field, J subgroup avian leucosis virus specific antigen epitope, fusion protein, specific antibody and its application are disclosed.Its amino acid sequence of J subgroup avian leucosis virus specific antigen epitope of the present invention such as SEQ ID NO：Shown in 1.Experiment shows that the epitope has reaction simultaneously reactionless to AB subgroup serum to J subgroup serum, being capable of specific recognition J subgroup avian leucosis virus.Fusion protein of the present invention can be made vaccine, and costimulatory receptor produces antibody, protection acceptor resistance J subgroup avian leucosis virus infection.Fusion protein immunization animal of the present invention can prepare J subgroup avian leucosis virus specific antibodies.J subgroup avian leucosis virus specific antibody of the present invention can develop detection kit, for distinguishing AB subgroups and J subgroup avian leucosis with J subgroup avian leucosis specific antigen epitope specific recognitions.

Description

J subgroup avian leucosis virus specific antigen epitope, fusion protein, specific antibody And its application
Technical field
The invention belongs to Animal diseases detection technique field, and in particular to J subgroup avian leucosis virus specific antigen tables Position, fusion protein, specific antibody and its application, especially J subgroup avian leucosis virus GP85 type specific antigens epitope, melt Hop protein, specific antibody and J subgroup avian leucosis detection kits.
Background technology
Avian leukosis is the birds ND caused by avian leukosis virus (ALV) and fowl sarcosis virus (ASV), It is great to China's poultry husbandry harm.ALV/ASV can be divided into 10 subgroups of A-J, and wherein A, B and J subgroup is to betide field chicken Exogenous virus.The current disease there is no effective treatment means, mainly control the disease to propagate by excluding positive chicken purification population. J hypotypes ALV can horizontal transmission and vertical transmission, vertical transmission can cause filial generation immune tolerance, and antibody test shows as feminine gender.Together When morbidity chicken there is acute tumor and cause death, once find that illness need to be immediately treated.AB hypotypes ALV mainly passes through vertical transmission Influence chicken quality, diseased chicken body weight increase is slow, and egg production is reduced.Diagnosis for J hypotypes and AB hypotypes ALV contributes to basis Mechanism and symptom that different subgroup virus cause carry out the prevention and control of avian leukosis, and chicken house also dependent on cost and demand to diseased chicken Processed in batches, reduced economic loss.
Epitope refers to have special construction and immunocompetent chemical group positioned at antigen molecule surface, can stimulate machine Body produces antibody or sensitized lymphocyte and the position that can be identified by it, also known as antigenic determinant, avian leukosis virus RNA codings Translate gag-pol-env in area.The nucleocapsid protein of gag gene code viral internal structures mainly has P27, P19, P15, P12 tetra- Albumen is planted, P27 albumen is highly conserved in avian leukosis poison, develops various avian leukosis detection reagents according to P27 albumen Box.But being only used for detection ALV viruses can not distinguish each virus subgroup.
Env is cyst membrane glycosylation surface protein, and Membrane surface proteins SU (surface glycoprotein) are formed after hydrolysis (gp85 gene codes) and transmembrane protein TM (transmembrane protein) (gp37 gene codes), gp85 determine virus Subgroup specificity and host range, are the main potential regions of epitope.At present to avian leukosis specific subset specificity The research work of epitope is all based on gp85 albumen to be carried out.
The method to the research of subgroup specific antigen epitope is confined to enter the specific subgroup gp85 albumen studied at present Row analysis, such as positioning J subgroups epitope is analyzed just for J subgroup gp85 albumen, and testing sample is J sub- in detection method But mouse monoclonal antibody prepared by group specificity serum or ALV-J albumen, J subgroups epitope that this method can find specificity are not By force, it is impossible to exclude the antigen recognition site that the epitopic regions for finding include other avian leukosis subgroups.With restructuring egg It is white when being made a distinction to different subtype avian leukosis virus infection serum, due to each subgroup gp85 albumen exist it is certain homologous Property, so in-vitro diagnosis cannot be carried out to avian leukosis antibody subtype.
The content of the invention
In view of this, it is an object of the invention to be directed to the defect of prior art, there is provided J subgroup specific antigen epitopes, Then exploitation detection kit is used to distinguish AB subgroups and J subgroup avian leucosis.
To realize the purpose of the present invention, the present invention is adopted the following technical scheme that：
The invention provides J subgroup avian leucosis virus specific antigen epitopes, its amino acid sequence such as SEQ ID NO：1 It is shown.
Applicant finds the otherness of AB subgroups and J subgroup gp85 albumen by comparing different subgroup gp85 protein sequences Region, is synthesized from distinct regions screening from 15 polypeptides of AB subgroups and J subgroups, is exactly found that can to distinguish fowl white The polypeptide of blood disease AB subgroups and J subgroups, while immunization experiment uses AB subgroups specific serum and J subgroups when analyzing 15 polypeptides Specific serum is sample, and such as certain J subgroups borne peptides has reaction simultaneously reactionless to AB subgroup serum as to J subgroup serum J subgroup specific antigen epitopes.For compared to previous studies, can find can distinguish AB subgroups and J subgroups specificity resist Former epitope.
Wherein, a kind of amino acid sequence of J subgroup avian leucosis virus specific antigen epitope is GTGGAEAELRDFIEKWKGDDHLRPYVNQSWTMVSPIN (J-Pep6, SEQ ID NO：2).Experiment shows, the epitope There is reaction simultaneously reactionless to AB subgroup serum to J subgroup serum, being capable of specific recognition J subgroup avian leucosis virus.
Present invention also offers the DNA molecular for encoding specific antigen epitope of the present invention.Due to the degeneracy of codon Property, there may be a variety of nucleotide sequences that can encode specific antigen epitope of the present invention.
Present invention also offers a kind of recombinant expression carrier, contain DNA molecular of the present invention.
On the other hand, present invention also offers a kind of engineering bacteria, it contains recombinant expression carrier of the present invention.
In the present invention, " engineering bacteria " is obtained by by the heat-shock transformed entrance host cell of recombinant expression carrier.This hair Expressed " host cell " includes protokaryon or eukaryotic in bright.
The host cell is selected from Escherichia coli, yeast, insect or mammalian cell.
In some embodiments, the host cell is Escherichia coli.
Specific antigen epitope of the present invention, DNA molecular, recombinant expression carrier or the engineering bacteria are all beneficial to The prevention or treatment of avian leukosis, thus the invention provides the specific antigen epitope, DNA molecular, recombinant expression carrier, Or application of the engineering bacteria in the medicine for preparing prevention or treatment avian leukosis.The specific antigen epitope, DNA point Son, recombinant expression carrier or the engineering bacteria can be used alone, and can also be used in combination, or with other known Drug combination.
Further, it is preferred that the medicine is vaccine.The impacted organ of patient can be applied directly to, also can whole body Administration.
Present invention also offers the fusion protein by the engineering bacterium expression.The engineering bacteria is expressed after induction, is separated Purifying can obtain the fusion protein of expression.
Fusion protein of the present invention being capable of costimulatory receptor generation antibody, protection acceptor resistance J subgroup avian leucosis virus Infection.Therefore, the fusion protein that the present invention is provided can be made vaccine for preventing the infection of J subgroup avian leucosis virus.
The present invention is also claimed a kind of vaccine, including fusion protein of the present invention.
Further, the vaccine also includes adjuvant.
Preferably, adjuvant is aluminum hydroxide adjuvant, Freund's complete adjuvant or incomplete Freund's adjuvant.
The present invention is also claimed J subgroup avian leucosis virus specific antibodies, and it is moved by the fusion protein immunization Monoclonal antibody or polyclonal antibody that thing is prepared from.
J subgroup avian leucosis virus specific antibody of the present invention can be with J subgroup avian leucosis specific antigen tables Position specific recognition, develops detection kit, for distinguishing AB subgroups and J subgroup avian leucosis.Therefore the invention provides institute State application of the J subgroup avian leucosis virus specific antibody in J subgroup avian leucosis detection kits are prepared.
Simultaneously present invention also offers a kind of J subgroup avian leucosis detection kit, it contains：
(1) specific antigen epitope of the present invention or fusion protein of the present invention；
And/or
(2) J subgroup avian leucosis virus specific antibody of the present invention.
Preferably, described J subgroup avian leucosis virus specific antibody can be monoclonal antibody, or many Clonal antibody.
Preferably, also including ELISA Plate in kit of the present invention.
It will be understood by those skilled in the art that fusion protein can be fixed on ELISA Plate and is used for by the kit that the present invention is provided J subgroup avian leucosis virus specific antibodies, also can be coated in ELISA Plate by J subgroup avian leucosis virus specific antibodies, use In detection fusion albumen.
Further, preferably, kit of the present invention also includes coating buffer, confining liquid, secondary antibody, nitrite ion, end Only liquid.
As shown from the above technical solution, the invention provides a kind of J subgroup avian leucosis virus specific antigen epitope, melt Hop protein, specific antibody and its application.Its amino acid sequence of J subgroup avian leucosis virus specific antigen epitope of the present invention Row such as SEQ ID NO：Shown in 1.Experiment shows that the epitope has reaction to J subgroup serum simultaneously to AB subgroup serum without anti- Should, being capable of specific recognition J subgroup avian leucosis virus.Fusion protein of the present invention can be made vaccine, and costimulatory receptor is produced Raw antibody, protection acceptor resistance J subgroup avian leucosis virus infection.It is sub- that fusion protein immunization animal of the present invention can prepare J Group's avian leukosis virus specific antibody.J subgroup avian leucosis virus specific antibody of the present invention can be white with J subgroup fowl The identification of blood disease-specific antigen epitope specificity, develops detection kit, for distinguishing AB subgroups and J subgroup avian leucosis.
Brief description of the drawings
In order to illustrate more clearly about the embodiment of the present invention or technical scheme of the prior art, below will be to embodiment or existing The accompanying drawing to be used needed for having technology description is briefly described.
Fig. 1 shows ALV-A, the gp85 protein sequence comparison charts of ALV-B, ALV-J virus；
Fig. 2 is the reaction effect of polypeptide and specific serum.
Specific embodiment
The invention discloses J subgroup avian leucosis virus specific antigen epitope and its application.Those skilled in the art can To use for reference present disclosure, technological parameter realization is suitably modified.In particular, all similar replacements and change are to this It is it will be apparent that they are considered as being included in the present invention for art personnel.The method of the present invention and product are It is described by preferred embodiment, related personnel can be not substantially being departed from present invention, spirit and scope to herein Described method is modified or suitably change is realized and apply the technology of the present invention with combining.
Below in conjunction with the embodiment of the present invention, the technical scheme in the embodiment of the present invention is clearly and completely described, Obviously, described embodiment is only a part of embodiment of the invention, rather than whole embodiments.Based in the present invention Embodiment, the every other embodiment that those of ordinary skill in the art are obtained under the premise of creative work is not made, all Belong to the scope of protection of the invention.
Unless otherwise specified, reagent involved in the embodiment of the present invention is commercially available prod, can be by business canal Road purchase is obtained.The reagent concrete configuration method that wherein ELISA immunologic detection methods are related to is：
Coating buffer solution (pH9.6 0.05M carbonate buffer solutions):Na2CO31.59g, NaHCO32.93g, distilled water 1000mL。
Lavation buffer solution (pH7.4PBST)：KH2PO40.2g, Na2HPO4·12H2O 2.9g, NaCl8.0g, KCl 0.2g, Tween-20 0.5mL, distilled water 1000mL.
Confining liquid:Bovine serum albumin(BSA) BSA 5g, lavation buffer solution 100mL.
The experimental technique of ELISA immune detections is：
(1) polypeptide coating
With the resuspended polypeptides of DMSO to 20mg/ml, polypeptide is diluted to 20ug/ml with coating buffer solution in 15ml centrifuge tubes, It is each to prepare 10ml.100 μ l polypeptide coating buffers are added to be placed in 4 DEG C of refrigerator overnights per hole in 96 hole elisa Plates.
(2) polypeptide coating plate is taken out, coating buffer is got rid of in pond, (the closing of the skimmed milks of 200 μ l 2% is then added per hole Liquid), with 37 degree of incubations 2 hours.
(3) deblocking liquid is got rid of in pond, is drained on blotting paper, then added per hole 100 μ l, 2% skimmed milks 50 × The chicken serum of dilution, 37 degree of incubators react 1 hour.
(4) serum deprivation sample is got rid of, per hole with 300 μ l cleaning solutions TBST respectively washing 3 times, is drained on filter paper, then add 100 μ l The HRP- sheep anti-chicken IgG ELIAS secondary antibodies of 5000x dilutions, 37 degree of incubators react 1 hour.
(5) ELIAS secondary antibody is got rid of, it is then ultrapure per the μ l of Kong Zaiyong 300 per hole with 300 μ l cleaning solutions TBST respectively washing 3 times Water washing is twice.After being drained on filter paper, 100 μ l tmb substrate solution are added per hole, wait substrate solution to react 20 minutes, flat board Take pictures.
(6) 50 μ l terminate liquids, extinction of the measurement in OD450 are added.
(7) judgement of positive findings, sample OD450 light absorption values are more than>(95% puts negative control average+2x standard deviations Letter is interval)
Embodiment 1, Peptide systhesis：
Obtain the ALV-A for representing popular strain on NCBI, ALV-B, ALV-J viral gp85 protein amino acid sequences, Protein sequence number is respectively：gi/559807359、gi/382933114、gi/350606570.Combined with ClustalX2 MEGA5.1 softwares are to these three subgroup virus gp85 albumen comparison results below figure 1.From ALV-A, ALV-B, ALV-J virus The rule that polypeptide is chosen in gp85 protein sequences is comparison result AB subgroups and the big region of J subgroup differences.Have selected more than 15 Peptide, wherein J hypotypes borne peptides totally 7, AB hypotypes borne peptides totally 8.The polypeptide sequence of synthesis see the table below 1, by polypeptide Solid phase synthetic instrument synthesizes, and carries out desalination preliminary purification.
The composition sequence of table 1 ALV-A, ALV-B, ALV-J subgroup specific polypeptide
Numbering Sequence Title Length Source
1 GYFFFQMILVCVVIISVVPGVGG J-Pep1 23 J
2 VNYILTIGVLVLCEVTGVRAD A-Pep2 21 A
3 NCTTLGTDRLVSSASITGGPDNSTT A-Pep3 25 A
4 DNSTTLTYRKVSCLLLKLNVSMWED A-Pep4 25 A
5 NGTNRTCVTFGSVCYEENSRSRVCHIFDGNFNGTG J-Pep5 35 J
6 GTGGAEAELRDFIEKWKGDDHLIRPYVNQSWTMVSPIN J-Pep6 38 J
7 PINTGSFSISS RYCGFTSNEARYYGGNLSDWCNSK J-Pep7 35 J
8 SNSTEPFTVVTADRHNLFTGSEYCGAYGYRFWE A-Pep8 33 A
9 VNQSREINETEPFSFTVNCTASNL A-Pep9 22 A
10 GGNCTAEWNYYAYGFTFGKQPEVLWNNGTA J-Pep10 30 J
11 SPNFTTWITYGPNITGHRRSR J-Pep11 21 J
12 APNHTDILKILANSSRTGIRRK AB-Pep12 21 AB
13 AIIPCIIKCFQDCLSRTMNQFMDDRIR J-Pep13 27 J
14 VCLPCLLQIVSRSVRKMVDN A-Pep14 23 A
15 NSIGYHTEYKKLQKACRQP A-Pep15 19 A
Embodiment 2, screening epitope cog region：
15 polypeptides are screened with AB subgroups and J subgroups specific serum, detects that polypeptide is imitated with indirect ElISA methods Really.Specifically detection method is：15 polypeptide DMSO for taking synthesis are resuspended to 20mg/ml, and pH9.6 carbonate buffer solutions are used respectively (coating buffer) is diluted to 20 μ g/ml coating CORNING ELISA Plates, and 4 DEG C overnight, is not coated with a group conduct and compares for negative and positive sex determination Standard；Plus 2% skimmed milk, 37 DEG C of closing 2h；Add 300 μ lPBST to clean per hole, residual in ELISA Plate is dried when pouring out liquid every time Moisture is simultaneously drained on clean blotting paper, is cleaned 5 times；Plus AB subgroups and the μ l/ holes of J subgroups specific serum 100, serum-dilution Multiple is 50 times, 250 times, 1000 times, and 37 DEG C are incubated 1 hour, clean 5 times；Plus 1：5000 times of horseradish peroxidases of dilution The μ l/ holes of goat-anti chicken IgY (IgG) ELIAS secondary antibody 100 of mark, 37 DEG C are incubated 1 hour, clean 5 times；Plus the μ l/ of TMB developers 100 Hole, it is static 10-15 minutes, plus ELIASA detection D after the μ l of terminate liquid 50450Value.The judgement of result is by detecting OD450>Standard female The standard deviation of the average+2 of sample × standard female sample carries out positive judgement, and statistically the result of determination can reach 95% confidential interval.
AB subgroup specific antigen epitopes are screened by AB subgroups and J subgroup specific serums using indirect elisa method It is No. 6 polypeptide (J-Pep6) GTGGAEAELRDFIEKWKGDDHLRPYVNQSWTMVSPIN in J subgroups source.
Fig. 2 results show No. 6 polypeptides to J subgroups specific serum difference extension rate sample have reaction and to AB subgroups Specific serum difference extension rate sample is substantially reactionless, illustrates No. 6 subgroup specificity of polypeptide, is J subgroup avian leucosis Epitope.
Embodiment 3, the specificity of detection epitope and susceptibility：
Screen to obtain by above-mentioned ELISA and there is specific polypeptide, meet J subgroups borne peptides only special to J subgroups Serum has reaction, and AB subgroups polypeptide only has reaction to AB subgroup specific serums.By with the preliminary mould of the reaction of field chicken serum Intend the Detection results of ELISA kit, totally 156 parts of sample：6 parts of standard female serum；IDEXX kits (like moral purchased from Beijing Shi Yuanheng bio tech ltd) detect 82 parts of the AB specific serums for obtaining；77 parts of J specific serums.
The specificity and susceptibility of epitope are detected with indirect ElISA methods：Polypeptide pH9.6 carbonate buffer solutions (bag By liquid) 20 μ g/ml coating CORNING ELISA Plates are diluted to, 4 DEG C are overnight；Plus 2% skimmed milk, 37 DEG C of closing 2h；Add 300 μ per hole LPBST is cleaned, and is dried when pouring out liquid every time and residual moisture and drained on clean blotting paper in ELISA Plate, is cleaned 5 times；Plus The μ l/ holes of 50 times of serum of above-mentioned dilution 100,37 DEG C are incubated 1 hour, clean 5 times；Plus 1：5000 times of horseradish peroxidases of dilution The μ l/ holes of goat-anti chicken IgY (IgG) ELIAS secondary antibody 100 of mark, 37 DEG C are incubated 1 hour, clean 5 times；Plus the μ l/ of TMB developers 100 Hole, it is static 10-15 minutes, plus ELIASA detection D after terminate liquid450Value.The judgement of result is by detecting OD450>Standard female sample The standard deviation of average+2 × standard female sample carry out positive judgement, statistically the result of determination can reach 95% Confidential interval.Specificity reaction polypeptide correctly judges the ratio of negative serum, and susceptibility reaction polypeptide correctly judges the ratio of the positive Rate.The results are shown in Table 2.Computing formula is as follows：
2 No. 6 susceptibility specificities of polypeptide (J-Pep6) of table
The result of table 2 shows that the susceptibility and specificity of No. 6 polypeptides (J-Pep6) reach more than 75%, is adapted to exploitation J subgroups Avian leukosis detection of specific antibody kit.
SEQUENCE LISTING
<110>China Agricultural University
<120>J subgroup avian leucosis virus specific antigen epitope, fusion protein, specific antibody and its application
<130> MP1702451
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 37
<212> PRT
<213>Artificial sequence
<400> 1
Gly Thr Gly Gly Ala Glu Ala Glu Leu Arg Asp Phe Ile Glu Lys Trp
1 5 10 15
Lys Gly Asp Asp His Leu Arg Pro Tyr Val Asn Gln Ser Trp Thr Met
20 25 30
Val Ser Pro Ile Asn
35

Claims (10)

1.J subgroup avian leucosis virus specific antigen epitopes, its amino acid sequence such as SEQ ID NO：Shown in 1.
2. the DNA molecular of specific antigen epitope described in claim 1 is encoded.
3. a kind of recombinant expression carrier, it contains nucleotide sequence described in claim 2.
4. a kind of engineering bacteria, it contains the recombinant expression carrier described in claim 3.
5. the weight described in the specific antigen epitope described in claim 1, the DNA molecular described in claim 2, claim 3 The application of group expression vector or engineering bacteria described in claim 4 in the medicine for preparing prevention or treatment avian leukosis.
6. application according to claim 5, it is characterised in that the medicine is vaccine.
7. as the fusion protein of engineering bacterium expression described in claim 4.
8.J subgroup avian leucosis virus specific antibodies, it is prepared from by fusion protein immunization animal described in claim 5 Monoclonal antibody or polyclonal antibody.
9. J subgroup avian leucosis virus specific antibody described in claim 8 is in J subgroup avian leucosis detection kits are prepared Application.
10. a kind of J subgroup avian leucosis detection kit, it contains：
(1) fusion protein described in the specific antigen epitope or claim 5 described in claim 1；
And/or
(2) the J subgroup avian leucosis virus specific antibodies described in claim 8.
CN201710183203.2A 2017-03-24 2017-03-24 J subgroup avian leucosis virus specific antigen epitope, fusion protein, specific antibody and its application Pending CN106866797A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201710183203.2A CN106866797A (en) 2017-03-24 2017-03-24 J subgroup avian leucosis virus specific antigen epitope, fusion protein, specific antibody and its application

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201710183203.2A CN106866797A (en) 2017-03-24 2017-03-24 J subgroup avian leucosis virus specific antigen epitope, fusion protein, specific antibody and its application

Publications (1)

Publication Number Publication Date
CN106866797A true CN106866797A (en) 2017-06-20

Family

ID=59172809

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201710183203.2A Pending CN106866797A (en) 2017-03-24 2017-03-24 J subgroup avian leucosis virus specific antigen epitope, fusion protein, specific antibody and its application

Country Status (1)

Country Link
CN (1) CN106866797A (en)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107353324A (en) * 2017-07-05 2017-11-17 河南牧业经济学院 The polypeptide being combined with avian leukosis virus p27 albumen and its application
CN110244039A (en) * 2019-05-05 2019-09-17 中国农业科学院哈尔滨兽医研究所（中国动物卫生与流行病学中心哈尔滨分中心） A kind of ELISA kit and its application detecting J subgroup avian leucosis virus antibody
CN110261599A (en) * 2019-05-16 2019-09-20 中国农业科学院哈尔滨兽医研究所（中国动物卫生与流行病学中心哈尔滨分中心） A kind of ELISA kit and its application detecting A, B subgroup avian leucosis virus antibody

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106442982A (en) * 2016-08-31 2017-02-22 扬州大学 Fowl leucovirus subgroup J specific antigenic polypeptide and application thereof

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106442982A (en) * 2016-08-31 2017-02-22 扬州大学 Fowl leucovirus subgroup J specific antigenic polypeptide and application thereof

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
DELONG LI等: "Avian leukosis virus subgroup A and B infection in wild birds of Northeast China", 《VETERINARY MICROBIOLOGY》 *
WEI PAN等: "Genetic diversity and phylogenetic analysis of glycoprotein GP85 of ALV-J isolates from Mainland China between 1999 and 2010: Coexistence of two extremely different subgroups in layers", 《VETERINARY MICROBIOLOGY》 *
XIAOFEI LI等: "Identification of a novel B-cell epitope specific for avian leukosis virus subgroup J gp85 protein", 《ARCH VIROL》 *
莫虹斐: "J亚群禽白血病病毒gp85单抗1E3柱抗原识别表位的初步分析", 《中国兽医杂志》 *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107353324A (en) * 2017-07-05 2017-11-17 河南牧业经济学院 The polypeptide being combined with avian leukosis virus p27 albumen and its application
CN107353324B (en) * 2017-07-05 2020-05-15 河南牧业经济学院 Polypeptide combined with avian leukosis virus p27 protein and application thereof
CN110244039A (en) * 2019-05-05 2019-09-17 中国农业科学院哈尔滨兽医研究所（中国动物卫生与流行病学中心哈尔滨分中心） A kind of ELISA kit and its application detecting J subgroup avian leucosis virus antibody
CN110261599A (en) * 2019-05-16 2019-09-20 中国农业科学院哈尔滨兽医研究所（中国动物卫生与流行病学中心哈尔滨分中心） A kind of ELISA kit and its application detecting A, B subgroup avian leucosis virus antibody
CN110261599B (en) * 2019-05-16 2022-02-08 中国农业科学院哈尔滨兽医研究所（中国动物卫生与流行病学中心哈尔滨分中心） ELISA kit for detecting A, B subgroup avian leukosis virus antibody and application thereof

Similar Documents

Publication Publication Date Title
CN113292640A (en) Novel recombinant coronavirus RBD trimer protein vaccine capable of generating broad-spectrum cross-neutralization activity, and preparation method and application thereof
JP5490725B2 (en) Binding protein and epitope blocking ELISA for universal detection of H5 subtype influenza virus
CN107033250B (en) Bovine coronavirus recombinant multi-epitope antigen and application thereof
US11376319B2 (en) Recombinant H7N9 subtype avian influenza virus, inactivated marked vaccine and preparation method thereof
CN103756973B (en) A kind of indirect ELISA testing kit of GCRV
CN109232734B (en) Monoclonal antibody specifically binding canine adenovirus, pharmaceutical composition, kit and application thereof
US20180346553A1 (en) Monoclonal antibody against novel epitopes of foot-and-mouth disease virus protein 3abc and uses thereof
CN109970852A (en) Secrete hybridoma cell strain and the application of rabies poison M protein monoclonal antibody
CN104450627B (en) The monoclonal antibody of 3 type envelope protein ORF132 of Cyprinus carpio herpesviruss, hybridoma cell strain and its application
CN106866797A (en) J subgroup avian leucosis virus specific antigen epitope, fusion protein, specific antibody and its application
CN109053883B (en) Binding protein of NS1 protein
CN109970851A (en) The preparation method of monoclonal antibody of CCV virus M protein and preparation method thereof, immunity colloidal gold test paper strip
KR20180038261A (en) An enzyme immuno assay kit for detecting H7N9 avian influenza virus
Zhang et al. Development and evaluation of a VP3-ELISA for the detection of goose and Muscovy duck parvovirus antibodies
CN106442981B (en) A kind of 1 type antibody indirect ELISA diagnostic kit of human bocavirus
CN110272488A (en) Feline calicivirus monoclonal antibody and its application
CN114874995A (en) Classical swine fever virus type 2E rns Monoclonal antibody hybridoma cell strain of protein and application
CN106866798A (en) J subgroup avian leucosis virus specific antigen epitope, fusion protein, specific antibody and its application
CN116284349B (en) Monkey pox antibody detection kit
CN109239341A (en) A kind of indirect ELISA reagent kit of ox Mannheimia haemolytica antibody test and its application
CN109762825A (en) A kind of the chicken virus mycoplasma antibody detection method and its dedicated kit of enzyme-linked aptamer
CN111848750A (en) Method and kit for rapidly enriching and detecting 2019-nCoV
McHugh et al. The detection of bornavirus in birds, mammals and snakes by sandwich ELISA of bornaviral matrix protein
KR20120028603A (en) Antibody for detecting hemagglutinin of swine influenza and use thereof
CN115925894B (en) Coronavirus monoclonal antibody and detection reagent prepared from same

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
RJ01 Rejection of invention patent application after publication

Application publication date: 20170620

RJ01 Rejection of invention patent application after publication