CN106995489A - A kind of Streptococcus suis truncated protein Sao and application - Google Patents
A kind of Streptococcus suis truncated protein Sao and application Download PDFInfo
- Publication number
- CN106995489A CN106995489A CN201610044176.6A CN201610044176A CN106995489A CN 106995489 A CN106995489 A CN 106995489A CN 201610044176 A CN201610044176 A CN 201610044176A CN 106995489 A CN106995489 A CN 106995489A
- Authority
- CN
- China
- Prior art keywords
- sao
- piglet
- glu
- lys
- truncated protein
- 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.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
- C07K14/315—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Streptococcus (G), e.g. Enterococci
-
- 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
- A61K39/02—Bacterial antigens
- A61K39/0208—Specific bacteria not otherwise provided for
-
- 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
- A61K39/02—Bacterial antigens
- A61K39/09—Lactobacillales, e.g. aerococcus, enterococcus, lactobacillus, lactococcus, streptococcus
- A61K39/092—Streptococcus
-
- 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
- A61K2039/55—Medicinal preparations containing antigens or antibodies characterised by the host/recipient, e.g. newborn with maternal antibodies
- A61K2039/552—Veterinary vaccine
-
- 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
- A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
- A61K2039/55511—Organic adjuvants
-
- 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
- A61K2039/70—Multivalent vaccine
Abstract
The invention discloses a kind of Streptococcus suis truncated protein Sao and application, described its amino acid sequence of truncated protein Sao is shown in SEQ ID NO.2.The protein stability is better than full-length proteins, it is not easy to degrade, and the Sao albumen provided using the present invention is made after vaccine, effectively can produce protective effect to piglet；Attacked using heavy dose of velogen strain SC19 after poison, its protection still reaches 60%.Sao albumen safety non-toxics, body temperature, breathing, appetite, the state of mind are all normal between the rear whole observation period immune to piglet and in-pig, and pregnant pig farrowing is normal, situations such as miscarriage, stillborn foetus do not occur.Sao albumen can also combine with other albumen is prepared into multiple vaccines, it is adaptable to a large amount of productions.
Description
Technical field
The present invention relates to domestic animal recombinant vaccine field, and in particular to a kind of Streptococcus suis truncated protein Sao and application.
Technical background
Streptococcus suis (Streptococcus suis, S.suis) is the main pathogen for causing Streptococcus suis, and S.suis points can be 33 serotypes, the respectively type of 1/2 type, 1~31 and 33 by the capsular polysaccharide (CPS) according to bacterium surface.Wherein streptococcus suis 2 (SS2) type is that virulence is most strong, prevalence is wide, is clinically separated frequency highest serotype.SS2 is a kind of important infecting both domestic animals and human encephalapthy agent, can cause pig meningitis, endocarditis, septicemia, arthritis, scrositis, pneumonia etc., often causes weanling pig or growing and fattening pigs die by visitation of God；The human infection bacterium, can cause meningitis, trigger permanent deafness, septicemia, endocarditis, or even dead.Guangdong Province is reported within 1991 first and finds the type streptococcus suspected case of pig 2, and season breaks out the disease to regional continuous 2 years of 1998-1999 Jiangsu Parts in full summer.Existing enough epidemiologic datas confirm that SS2 constitutes serious threat to the aquaculture and people's health of China at present.Therefore, the infection and control Streptococcus suis for developing effective vaccine to prevent Streptococcus suis are extremely urgent.
Subunit seedling has the advantages that the security of the immune efficacy height of live vaccine and inactivated vaccine is good, Yuanyi Li et al. confirm that Sao is the surface membrane protein of Streptococcus suis, Sao specific antibody can have good immunogenic response with 25 plants in the 28 serological type strain lysates and 26 plants of SS2 in 33 serotypes, illustrate that Sao has high conservative in Streptococcus suis.
In addition, the immune protective effect and adjuvant of Sao albumen have much relations.Although Yuanyi Li et al. have found that Sao and adjuvant E mulsigen-Plus is used in mixed way can cause higher antibody level afterwards, piglet can not be protected to resist the infection of Streptococcus suis.Adjuvant Emulsigen-Plus was substituted for adjuvant Quil A by the researcher later, can but provide the protection of piglet more than 80%.K.-J.Hsueh et al. confirms that Sao and commercialization adjuvant w/o/w adjuvants can also provide 80% protection to mouse.
Sao is divided into 3 types according to the difference of gene order length：Sao-S (1.5kb), Sao-M (1.7kb), Sao-L (2.0k b), wherein Sao-M is distributed most wide in bacterial strain.Sao-M full-length proteins have 110KD, Wang Jing et al. to find that total length sao protein purifications difficulty is big, and protein stability is poor, degradation speed block, and these factors hinder the further research and development of Sao protein vaccines.
The content of the invention
Object of the present invention is to provide a kind of Streptococcus suis truncated protein Sao, its sequence is shown in SEQ ID NO.2.Its corresponding nucleotides sequence is classified as shown in SEQ ID NO.1.The truncated protein Sao that the present invention is provided is stability albumen, is dissolved in PBS cushioning liquid, is stored in -80 DEG C and takes out after one month and does not find that albumen has signs of degradation.
It is another object of the present invention to provide applications of the Streptococcus suis truncated protein Sao in streptococcus suis infection vaccine is prepared; truncated protein Sao and adjuvant are prepared into vaccine; poison is attacked after intramuscular immunisation piglet, booster immunization, higher protection can be provided to piglet.
Last purpose of the invention is the provision of applications of the truncated protein Sao in multiple vaccines are prepared, and the truncated protein Sao that the present invention is provided is prepared into multiple vaccines with other protein vaccines, can play the effect of a pin multiple-effect.
In order to achieve the above object, the present invention takes following technical measures：
Full-length proteins Sao is unstable, easily degraded, applicant is effectively truncated total length Sao albumen, protein sequence after truncation is shown in SEQ ID NO.2, truncated protein Sao is changed into stability albumen, it is dissolved in PBS cushioning liquid, is stored in -80 DEG C and takes out after one month and do not find that albumen has signs of degradation.
Applications of the Streptococcus suis truncated protein Sao in streptococcus suis infection vaccine is prepared, including truncated protein Sao and adjuvant are prepared by mixing into vaccine in proportion, intramuscular immunisation piglet can effectively protect piglet to resist the infection of Streptococcus suis.
Applications of the truncated protein Sao in multiple vaccines are prepared; the truncated protein Sao that the present invention is provided is prepared into multiple vaccines with other protein vaccines; it is preferred that truncated protein Sao and erysipelothrix ruhsiopathiae surface truncated protein SpaA (shown in SEQID NO.3) are prepared into combined vaccine; intramuscular immunisation piglet; it can effectively improve and attack piglet survival rate after poison, two immune protective effects being independent of each other each other to piglet.
Specific method refers to embodiment.
Compared with prior art, the present invention has advantages below：
1. the Sao property of protein that the present invention is provided is stable, it is not easy to degrades, better than full-length proteins, is prepared into vaccine rear stability good, it is adaptable to a large amount of productions.
2. the Sao albumen provided using the present invention is made after vaccine, protective effect effectively can be produced to piglet, be attacked using heavy dose of velogen strain SC19 after poison, its protection still reaches 60%.
3. the Sao albumen safety non-toxics that the present invention is provided, body temperature, breathing, appetite, the state of mind are all normal between the rear whole observation period immune to piglet and in-pig, and pregnant pig farrowing is normal, situations such as miscarriage, stillborn foetus do not occur.
4. the Sao albumen that the present invention is provided can also combine with other albumen is prepared into multiple vaccines, the effect of a pin multiple-effect can be played.
Brief description of the drawings
Fig. 1 is the antibody level schematic diagram that mouse is immunized in truncated protein Sao.
Fig. 2 is to attack malicious protection effect diagram after mouse is immunized in truncated protein Sao.
Fig. 3 is that Temperature changing schematic diagram after piglet is immunized in truncated protein Sao.
Fig. 4 is that antibody level schematic diagram after piglet is immunized in truncated protein Sao.
Fig. 5 is to attack malicious protection effect diagram after piglet is immunized in truncated protein Sao.
Fig. 6 is that Temperature changing schematic diagram after piglet is immunized in joint seedling.
Fig. 7 is that truncated protein Sao antibody level schematic diagrames after piglet are immunized in joint seedling.
Fig. 8 is that truncated protein SpaA antibody level schematic diagrames after piglet are immunized in joint seedling
Embodiment
Reagent of the present invention if not otherwise specified, is purchased from commercial channel or the general configuration reagent for this area.The technical scheme is the conventional scheme of this area if not otherwise specified.
Embodiment 1：
It is prepared by Streptococcus suis truncated protein Sao synthesis and vaccine：
1.1 truncated protein Sao synthesis
Truncated protein Sao sequence is synthesized as shown in SEQ ID NO.2 by Sangon Biotech (Shanghai) Co., Ltd..Truncated protein Sao Purities are 99%, every 5mg after synthesis, are white powder, are kept in dark place in -20 degree, are completely soluble in water.By the protein dissolution in PBS cushioning liquid, it is stored in -80 DEG C and takes out after one month and do not find that albumen has signs of degradation.
Before use, with sterile PBS (NaCl 8.0g, KCl 0.2g, KH2PO40.24g, Na2HPO4×12H2O 3.628g, pH7.4) it is diluted by prescribed concentration.Preheated using preposition in 37 degree of incubators.
The preparation of 1.2 vaccines
Formulation：Water-in-oil type (W/O)
Adjuvant is constituted：White Mineral Oil (Marcol52), Tween 80 (CRILLET4), sorbester p17 (CRILL4).
Oil phase：White Mineral Oil be heated to it is transparent after and sorbester p17 by volume 94：6 prepare.
Aqueous phase：Albumen is constituted with Tween 80, and Tween 80 accounts for 0.4% (volume ratio) of aqueous phase.
Vaccine is emulsified：Aqueous phase presses 1 with oil phase：1 volume mixture is emulsified.Make truncated protein Sao final concentration of 1mg/ml, for following experiment.
1.3 routine inspection
Vaccine character is detected：Vaccine is instilled in clear water after emulsification, and the first drop is scattered, and the second oil dripping pearl rocks liquid level oil droplet and is not demulsified.Sterility testing：A little vaccine coating TSA plates are taken, 37 DEG C of incubator cultures are grown for 18 hours without bacterium colony.
Stability test：3000rpm/min is centrifuged 15 minutes, judges whether emulsification stablizes according to separation situation, and vaccine is emulsified after centrifugation and is not layered.
By protein dissolution in PBS cushioning liquid, -80 DEG C preserve 1 month, 3 months, 6 months respectively, do not find that property of protein changes.
The good vaccine of above-mentioned emulsification is placed in into 4 DEG C to preserve 1 month, 3 months, 6 months respectively, vaccine Character change is not detected by.Embodiment 2：
Applications of the truncated protein vaccine Sao in streptococcus suis infection vaccine is prepared
1st, the immune protective effect of vaccine is evaluated on mouse model
1.1 experimental program
Experimental animal：4 week old female BAl BIcs/c small white mouses, purchased from Disease Prevention Control Center, Hubei Prov.
Attack toxic bacterial strain：Streptococcus suis 2-type SC19.
Experiment packet：Body weight 16g female BAl BIcs/c small white mouses 20 are taken, 2 groups are randomly divided into:Vaccine I groups (Sao), control group (adjuvant and PBS), every group 10.Docking takes blood weekly between the immune later stage, the monitoring for serum specific antibody.Immunization method：The dose subcutaneous immunization experiment mouse of vaccine prepared by embodiment 1 by 100 μ l/ only；Booster immunization 1 time after 2 weeks (immunizing dose and position are with immune for the first time).
The detection of 1.2 serum specific antibodies
Elisa plate is coated with overnight in 4 DEG C using truncated protein Sao100ng/ holes, and the antibody titer of serum is separated with the method detection of indirect ELISA；As a result Fig. 1 is seen.
Detection discovery is carried out to the antibody level of mouse after immune：Higher antibody level can be just produced with inducing mouse within first week after first immunisation, there is 27.When two exempt from 14 days, mouse internal antibody level reaches a higher level, and the Mean antibody titer of positive serum can reach 215。
1.3 immunized mices, which are attacked, protects situation after poison
Two exempt from challenge test after 14 days, and it is 3.3 × 10 to attack toxic agent amount8CFU/ is only.Continuous Observation one week, records the Clinical symptoms and death condition of mouse.
As a result it is as follows：
SC19 plants of absolute lethal bacterium amount LD100 is 3.0 × 108CFU/ only, attack malicious result and see Fig. 2, and mouse survival rate is 70%, and 7 mouse of survival are without clinical symptoms by immune mouse；And control group mice is all dead, survival rate is 0.It can be seen that the vaccine on mouse that after Sao protein truncations plus adjuvant is made has higher protecting effect by antibody level and immune group mouse survival rate, the immunogenicity of the albumen does not disappear.
2nd, the immune protective effect of vaccine is evaluated on piglet model
2.1 experimental program
Experimental animal：The negative weanling pig of 4 week old Streptococcus suis detection, purchased from Wuhan economic cycle green grass lake Tian Zhong pig farms.
Attack toxic bacterial strain：Streptococcus suis 2-type SC19.
Experiment packet：4 week old weanling pigs are divided into 3 groups, 1 group (Sao), immune 2 groups (Sao), control group (adjuvant and PBS) is immunized, 1 group and control group every group 10 is immunized, it is immune 2 groups 5, immune 1 group, adjuvant and PBS mixing control group are used to attack poison, immune 2 groups are used to detect antibody level, continuous detection 8 months.
Immunization wayses：Buttocks muscles multi-point injection, head exempt from after every 14 days booster immunizations once (immunizing dose and position are exempted from head), immunizing dose be 2ml/ heads.
The detection of 2.2 serum specific antibodies
Ear vein takes blood weekly for immune 1 group, adjuvant and PBS mixing control group, the detection for serum specific antibody；Immune 2 groups of every months take a blood sample once, the detection for serum specific antibody.Elisa plate is coated with using the truncated protein Sao100ng/ holes of purifying, 4 DEG C overnight.The antibody level in piglet body is detected with indirect ELISA.
2.3 immune rear piglet Temperature changing detections
Body temperature, continuous detection 7 days, the feeding of the immune rear piglet of observation and mental condition are monitored after first immunisation daily.
2.4 immune swines, which are attacked, protects situation after poison
Two exempt to carry out challenge test after 14 days, and it is 1.8 × 10 to attack toxic agent amount6The SC19 of cfu/, is 1.5 times of piglet lethal dose LD100.Continuous Observation 14 days, record Clinical symptoms and death condition.
2.5 immune effects are assessed
Piglet does not have clinical onset symptom after immune, second day body temperature can rise after immune 1 group, control group is immune, ascensional range is smaller, the symptom such as have no piglet and have apocleisis, greedy sleeping, spirit depressed, third body temperature is returned to normal, period piglet feed intake has no reduction, and the of short duration rising of body temperature is smaller to the growth effect of piglet, and Temperature changing is shown in Fig. 3.
Antibody level detection after piglet immunological Sao is shown：The antibody level of Sao induction piglets gradually rises, and 4th week is up to 2 after head exempts from20Left and right, sees Fig. 4；Attack after poison, immune 1 group there are 4 piglets dead in 7 days, survives 6, sees Fig. 5.Health survival piglet is sentenced into euthanasia and carries out cut open inspection point bacterium, kidney,spleen,liver, the heart are not all separated to SC19.Control group piglet falls ill successively, back leg arthroncus, and spirit is depressed, and happiness is sleeping, conjunctivitis, strikes in swimming shape, has piglet dead successively, last all dead complete.The experimental result of body animal further illustrates that truncated protein Sao prepared by the present invention can provide preferable protecting effect for piglet.
Start within 2 weeks to detect antibody after antibody level detection to immune 2 groups of carry out 8 months, booster immunization, testing result shows：Head exempt from after the 28th day~90 days antibody titer average values 220Left and right；Exempt from rear 4th month to 6th month antibody titer from head to be gradually reduced, antibody level is still higher, and the antibody titer average value of the 6th month is 210, still there is protection left and right to piglet, illustrates that antibody level can at least be maintained 6 months.Though 210 days can detect antibody, antibody level is relatively low, and antibody titer average value only has 25。
Embodiment 3：Safety examination
In order to examine prepared vaccine to the safety effects of piglet and in-pig, safety examination is carried out to 35 age in days piglets and pregnant 70 age in days sow.
3.1 experimental program：
1) by the vaccine of preparation respectively by the age in days sodium selenite of buttocks muscles branch injection inoculation 35, every batch of vaccine injection 5, every inoculation 2ml observes its clinical manifestation and determines body temperature.
2) vaccine prepared is respectively by the buttocks muscles branch injection inoculation gestation healthy sow of 70 days, and every batch of vaccine injection 5, every inoculation 2ml observes its clinical manifestation and farrowing situation.
2. result of the test
3.2.1 the security that subunit vaccine of the invention is inoculated with to weanling pig
After the healthy weanling pig of the age in days of vaccine inoculation 35 of preparation, every batch of vaccine injection 5, piglet breathes situation, mental status, appetite etc. normally, and simply second day body temperature of vaccine inoculation slightly rises, and third body temperature declines recover normally rapidly.Illustrate that the vaccine product is fine to weanling pig security, Temperature changing is shown in (table 1).
Mean body temperature change after the age in days weanling pig of vaccine inoculation 35 prepared by the present invention of table 1
3.2.2 the security (produce surviving of son situation) that subunit vaccine of the invention is inoculated with to pregnant animal (in-pig)
As a result the pregnant sow on the 70th of health of 3 batches of subunit vaccine inoculations of the invention body temperature, breathing, appetite, state of mind between the whole observation period is all normal (table 2), and farrowing is normal, situations such as miscarriage, stillborn foetus do not occur (table 3).
Mean body temperature change after the healthy in-pig of vaccine inoculation of the present invention of table 2
The healthy in-pig inoculation of table 3 prepares the farrowing situation after vaccine
Embodiment 4：
Applications of the truncated protein Sao in multiple vaccines are prepared
1. the preparation of combined vaccine
Prepare truncated protein Sao and erysipelothrix ruhsiopathiae surface truncated protein SpaA joint seedling by the method in embodiment 1, Sao and SpaA content are respectively 1mg/ml.
Described erysipelothrix ruhsiopathiae surface truncated protein SpaA amino acid sequence is shown in SEQ ID NO.3, truncated protein SpaA used in the present invention is genetic engineering bacterium Escherichia coli (Escherichia coli) BL21/pET-28a-spaA, and deposit number is CCTCC NO：M2015076 secretions are obtained, and described bacterial strain delivered to China typical culture collection center preservation, address on 2 2nd, 2015：Wuhan, China Wuhan University, Classification And Nomenclature：Escherichia coli BL21/pET28a-spaA.
2. combine seedling immune effect on piglet
1) experimental program
Experimental animal：The negative weanling pig of 4 week old Streptococcus suis detection, purchased from Wuhan economic cycle green grass lake Tian Zhong pig farms.
Attack toxic bacterial strain：Streptococcus suis 2-type SC19, erysipelothrix ruhsiopathiae SE38 (abbreviation SE38, Li Jingtao, Hubei some areas erysipelothrix ruhsiopathiae separation strains drug resistance analysis is cultivated and feed [J] .2015.2,11-14), it is 1.8 × 10 that SC19, which attacks toxic agent amount,6CFU/ heads, are 1.5 times of lethal dose ID100；It is 9.0 × 10 that SE38, which attacks toxic agent amount,8CFU/ heads, are 3 times of lethal dose LD100.
Experiment packet：4 week old weanling pigs are divided into 4 groups, and 1 group (Sao+SpaA), immune 2 groups (Sao+SpaA), 2 groups of control group (adjuvant+PBS), every group each 10 is immunized.
Immunization wayses：Buttocks muscles multi-point injection, head exempt from after every 14 days booster immunizations once, immunizing dose be 2ml/ heads.
2) detection of serum specific antibody
Randomly selecting piglet in 4 experimental groups, ear vein takes blood weekly, the detection for serum specific antibody.Elisa plate is coated with using the truncated protein Sao100ng/ holes of purifying, 4 DEG C overnight, with Sao antibody levels in indirect ELISA detection piglet separation serum.Elisa plate is coated with using the truncated protein SpaA100ng/ holes of purifying, 4 DEG C overnight, with SpaA antibody levels in indirect ELISA detection piglet separation serum.
3) piglet clinical observation after being immunized
Body temperature, continuous detection 7 days, the feeding of the immune rear piglet of observation and mental condition are monitored after first immunisation daily.
4) immune swine, which is attacked, protects situation after poison
Two exempt to carry out challenge viral dosage after 14 days, Continuous Observation 14 days, record Clinical symptoms and death condition.
5) immune effect is assessed
Piglet does not have clinical onset symptom after immune, second day body temperature of piglet can slightly rise after immune, the symptom such as have no piglet and have apocleisis, greedy sleeping, spirit depressed, third body temperature is returned to normal, period piglet feed intake has no reduction, the of short duration rising of body temperature is smaller to the growth effect of piglet, and Temperature changing is shown in Fig. 6.
Sao and SpaA antibody levels detection after piglet immunological is shown：After immune piglet, antibody titer gradually rises, and the antibody titer of piglet can reach 2 at the end of exempting from 2 weeks to two20Left and right.Increase with the antibody level of monomer vaccine immunity and compare, the antibody growth that piglet is immunized in joint seedling has no abnormal changes, and explanation can be used in combination, and be not in the phenomenon that antibody is offset, see Fig. 7 and Fig. 8.
Attack poison and the results are shown in Table 4, truncated protein Sao seedlings can provide the protection of piglet 60%, and SpaA can provide 90% protection to piglet.This experimental result illustrates that the joint seedling further prepared does not interfere with protecting effect each other to piglet.
The result that the joint seedling of table 4 is attacked after poison
Truncated protein Sao prepared by the present invention, relative to full-length proteins, is stability albumen, and can provide higher protecting effect after adjuvant mixing to piglet, illustrates that its immunogenicity is not reduced.Combined vaccine is prepared by mixing into erysipelothrix ruhsiopathiae truncated protein SpaA piglet is immunized, be not in the phenomenon that antibody is offset.It is described above：Individually truncate the infection that Sao albumen seedling can protect piglet to resist the strong poison SC19 of streptococcus suis 2-type; the infection that joint seedling can also protect piglet to resist Streptococcus suis or erysipelothrix ruhsiopathiae is constituted with truncated protein SpaA; meet the preventative strategies of current " the anti-many diseases of a pin ", with very important actual application value.
SEQUENCE LISTING
<110>Hua Zhong Agriculture University
<120>A kind of Streptococcus suis truncated protein Sao and application
<130>A kind of Streptococcus suis truncated protein Sao and application
<160> 3
<170> PatentIn version 3.1
<210> 1
<211> 777
<212> DNA
<213>Artificial sequence
<400> 1
atgacaacct gatgggggac aggctacttc aaaggcggtt aatgtcaaaa
taccagcagt 60
agtacgacta tttggtcgtg agcttctaga aaatgaattt aaatttgagc
ttagagaagc 120
gaatggcgag gaactccctg tccttgatac agctcaaaat acaaaagagg
gtcaagttag 180
atttaaaaat ctatcattcg ataagcctgg caaatactgg tatacaattt
cagaagtaaa 240
agatgagctt ggtggtattg agtatgattc gaaatatatt gtagcaaaaa
taactgtaga 300
agatcgaaac gggcaattac aggcaatgat cgaatttatt gataatgaca
atgtctttaa 360
caatttctat acacctgctc cagctgctgc tagtctttcg ataaaaaaag
tcctcgaggg 420
acgtacctta aacaccggtg aattcgaatt tgttttaaaa aatgaaaaag
gcgatgaaat 480
cgaaaaggta agcaatcaag cagatggttc tgtaaacttt agtgccctaa
catttacaaa 540
agagggaacc tatacctaca ctgtttcaga agttgatggt ggacttggcg
atattatcta 600
tgacaaatca gatattaagg ccactgttac tgtgaaagat aacaatcacg
gacaactagt 660
ctcaacagtg acttatgaaa atagcgatca aatcttcgag aatattttga
atcctgggaa 720
gttaatagcg ccaaccacgg atagcgttat tactgataat gaagtctcta
aggaagc 777
<210> 2
<211> 258
<212> PRT
<213>Artificial sequence
<400> 2
Met Asn Thr Lys Lys Trp Arg Thr Ser Leu Leu Ile Pro Gly Ile Val
1
5
10
15
Leu Phe Gly Thr Val Ala Leu Val Asn Asn Val Ser Ala Gln Glu Val
20
25
30
Lys Asn Thr Ile Ile Ser Ala Lys Gln Pro Asp Gly Gly Gln Ala Thr
35
40
45
Ser Lys Ala Val Asn Val Lys Ile Pro Ala Val Val Arg Leu Phe Gly
50
55
60
Arg Glu Leu Leu Glu Asn Glu Phe Lys Phe Glu Leu Arg Glu Ala Asn
65
70
75
80
Gly Glu Glu Leu Pro Val Leu Asp Thr Ala Gln Asn Thr Lys Glu Gly
85
90
95
Gln Val Arg Phe Lys Asn Leu Ser Phe Asp Lys Pro Gly Lys Tyr Trp
100
105
110
Tyr Thr Ile Ser Glu Val Lys Asp Glu Leu Gly Gly Ile Glu Tyr Asp
115
120
125
Ser Lys Tyr Ile Val Ala Lys Ile Thr Val Glu Asp Arg Asn Gly Gln
130
135
140
Leu Gln Ala Met Ile Glu Phe Ile Asp Asn Asp Asn Val Phe Asn Asn
145
150
155
160
Phe Tyr Thr Pro Ala Pro Ala Ala Ala Ser Leu Ser Ile Lys Lys Val
165
170
175
Leu Glu Gly Arg Thr Leu Asn Thr Gly Glu Phe Glu Phe Val Leu Lys
180
185
190
Asn Glu Lys Gly Asp Glu Ile Glu Lys Val Ser Asn Gln Ala Asp Gly
195
200
205
Ser Val Asn Phe Ser Ala Leu Thr Phe Thr Lys Glu Gly Thr Tyr Thr
210
215
220
Tyr Thr Val Ser Glu Val Asp Gly Gly Leu Gly Asp Ile Ile Tyr Asp
225
230
235
240
Lys Ser Asp Ile Lys Ala Thr Val Thr Val Lys Asp Asn Asn His Gly
245
250
255
Gln Leu
<210> 3
<211> 450
<212> PRT
<213>Artificial sequence
<400> 3
Met Asp Ser Thr Asp Ile Ser Val Ile Pro Leu Ile Gly Glu Gln Val
1
5
10
15
Gly Leu Leu Pro Val Leu Pro Gly Thr Gly Val His Ala Gln Glu Tyr
20
25
30
Asn Lys Met Thr Asp Ala Tyr Ile Glu Lys Leu Val Ser Leu Ile Asn
35
40
45
Gln Lys Val Lys Pro Phe Leu Ile Asn Glu Pro Lys Gly Tyr Gln Ser
50
55
60
Phe Glu Ala Val Asn Glu Glu Ile Asn Ser Ile Val Ser Glu Leu Lys
65
70
75
80
Asn Glu Gly Met Ser Leu Gln Asn Ile His His Met Phe Lys Gln Ser
85
90
95
Ile Gln Asn Leu Ala Thr Arg Ile Gly Tyr Arg Ser Phe Met Gln Asp
100
105
110
Ala Met Tyr Leu Glu Asn Phe Glu Arg Leu Thr Ile Pro Glu Leu Asp
115
120
125
Glu Ala Tyr Val Asp Leu Leu Val Asn Tyr Glu Val Lys His Arg Ile
130
135
140
Leu Val Lys Tyr Glu Gly Lys Val Lys Gly Arg Ala Pro Leu Glu Ala
145
150
155
160
Phe Ile Val Pro Leu Arg Asp Arg Ile Arg Ser Met Asn Glu Met Ala
165
170
175
Ala Glu Val Asn Gly Leu Pro Glu Ala His Glu Asp Phe Leu Val Ser
180
185
190
Asp Ser Ser Glu Tyr Asn Asp Lys Leu Asn Asn Ile Asn Phe Ala Leu
195
200
205
Gly Leu Gly Val Ser Glu Phe Ile Asp Tyr Asn Arg Leu Glu Asn Met
210
215
220
Met Glu Lys Glu Ile His Pro Leu Tyr Leu Glu Leu Tyr Ala Met Arg
225
230
235
240
Arg Asn Arg Gln Ile Gln Val Val Arg Asp Val Tyr Pro Asn Leu Glu
245
250
255
Arg Ala Asn Ala Val Val Glu Ser Leu Lys Thr Ile Lys Asp Ile Lys
260
265
270
Gln Arg Gly Lys Lys Leu Gln Glu Leu Leu Glu Ile Tyr Ile Gln Arg
275
280
285
Ser Gly Asp Val Arg Lys Pro Asp Val Leu Gln Arg Phe Ile Gly Lys
290
295
300
Tyr Gln Ser Val Val Asp Glu Glu Lys Asn Lys Leu Gln Asp Tyr Leu
305
310
315
320
Glu Ser Asp Ile Phe Asp Ser Tyr Ser Val Asp Gly Glu Lys Ile Arg
325
330
335
Asn Lys Glu Ile Thr Leu Ile Asn Arg Asp Ala Tyr Leu Ser Met Ile
340
345
350
Tyr Arg Ala Gln Ser Ile Ser Glu Ile Lys Thr Ile Arg Ala Asp Leu
355
360
365
Glu Ser Leu Val Lys Ser Phe Gln Asn Glu Glu Ser Asp Ser Lys Val
370
375
380
Glu Pro Glu Ser Pro Val Lys Val Glu Lys Pro Val Asp Glu Glu Lys
385
390
395
400
Pro Lys Asp Gln Lys Lys Leu Val Asp Gln Ser Lys Pro Glu Ser Asn
405
410
415
Ser Lys Glu Gly Trp Ile Lys Lys Asp Asn Lys Trp Phe Tyr Ile Glu
420
425
430
Lys Ser Gly Gly Met Ala Thr Gly Trp Lys Lys Val Ala Asp Lys Trp
435
440
445
Pro Tyr
450
Claims (5)
1. a kind of Streptococcus suis truncated protein Sao, its sequence is shown in SEQ ID NO.2.
2. the corresponding nucleotide sequence of albumen described in claim 1.
3. application of the albumen described in claim 1 in streptococcus suis infection vaccine is prepared.
4. application of the albumen described in claim 1 in multiple vaccines are prepared.
5. multiple vaccines according to claim 5, described multiple vaccines are to include the multiple vaccines of amino acid shown in the amino acid and SEQ ID NO.3 shown in SEQ ID NO.2.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610044176.6A CN106995489B (en) | 2016-01-22 | 2016-01-22 | A kind of Streptococcus suis truncated protein Sao and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610044176.6A CN106995489B (en) | 2016-01-22 | 2016-01-22 | A kind of Streptococcus suis truncated protein Sao and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106995489A true CN106995489A (en) | 2017-08-01 |
| CN106995489B CN106995489B (en) | 2018-11-06 |
Family
ID=59428811
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610044176.6A Active CN106995489B (en) | 2016-01-22 | 2016-01-22 | A kind of Streptococcus suis truncated protein Sao and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106995489B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111378015A (en) * | 2019-12-27 | 2020-07-07 | 无锡市妇幼保健院 | Synthetic peptide for detecting S.suis2 infection and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101031647A (en) * | 2004-02-27 | 2007-09-05 | 财团法人化学及血清疗法研究所 | Process for producing erysipelothrix rhusiopathiae surface protective antigen mutant in escherichia coli |
| CN101313068A (en) * | 2005-09-02 | 2008-11-26 | 蒙特利尔大学 | Streptococcus suis polypeptides and polynucleotides encoding same and their use in vaccinal and diagnostic applications |
-
2016
- 2016-01-22 CN CN201610044176.6A patent/CN106995489B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101031647A (en) * | 2004-02-27 | 2007-09-05 | 财团法人化学及血清疗法研究所 | Process for producing erysipelothrix rhusiopathiae surface protective antigen mutant in escherichia coli |
| CN101313068A (en) * | 2005-09-02 | 2008-11-26 | 蒙特利尔大学 | Streptococcus suis polypeptides and polynucleotides encoding same and their use in vaccinal and diagnostic applications |
| CN103483431A (en) * | 2005-09-02 | 2014-01-01 | 蒙特利尔大学 | Streptococcus suis polypeptides and polynucleotides encoding same and their use in vaccinal and diagnostic applications |
Non-Patent Citations (3)
| Title |
|---|
| HSUEH KJ, LEE JW, HOU SM, CHEN HS, CHANG TC, CHU CY: "Evaluation on a Streptococcus suis vaccine using recombinant sao-l protein manufactured by bioreactors as the antigen in pigs", 《TRANSBOUNDARY AND EMERGING DISEASES》 * |
| YU,X., QIAN,X., LI,R., QU,T. AND HU,Y.: "AGM15970.1", 《GENBANK》 * |
| 吾鲁木汗·那孜尔别克 等: "猪丹毒丝菌天然SpaA和重组SpaA-N免疫保护效果的评价", 《微生物学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111378015A (en) * | 2019-12-27 | 2020-07-07 | 无锡市妇幼保健院 | Synthetic peptide for detecting S.suis2 infection and application thereof |
| CN111378015B (en) * | 2019-12-27 | 2021-01-01 | 无锡市妇幼保健院 | Synthetic peptide for detecting S.suis 2 infection and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106995489B (en) | 2018-11-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104862286B (en) | Porcine pseudorabies virus gene-deleted strain, vaccine composition and its preparation method and application | |
| JP6475741B2 (en) | Herpesvirus subunit vaccine and its production method and application | |
| CN104784686B (en) | TGEV, PEDV bigeminal live vaccine and preparation method thereof | |
| CN104513827A (en) | Porcine epizootic diarrhea virus strain, attenuated vaccine strain thereof and application thereof | |
| CN103725651B (en) | One plant of Porcine epidemic diarrhea virus and its application | |
| CN106148287B (en) | Porcine epidemic diarrhea virus strain and vaccine composition, preparation method and application thereof | |
| CN104004774A (en) | Swine pseudorabies virus and vaccine composition as well as preparation method and application thereof | |
| CN105462936B (en) | A kind of Porcine epidemic diarrhea virus MY01 plants and its application | |
| CN102851257A (en) | Attenuated vaccine strain for avian infectious bronchitis virus and application thereof | |
| CN108486067A (en) | The inactivated vaccine and application of Porcine epidemic diarrhea virus variant and its preparation | |
| CN110387355A (en) | Express recombinant porcine pseudorabies Strain, the vaccine and its preparation method and application of pig circular ring virus Cap protein gene | |
| CN108823218A (en) | Chicken infectivity bursa of Fabricius virus VP 2 gene, its expression product, its subunit vaccine and application | |
| CN105018433A (en) | Porcine pseudorabies virus gene deletion strain, vaccine composition and preparation method and application thereof | |
| CN113355287A (en) | Bivalent vaccine for porcine circovirus type 2 and type 3 and preparation method thereof | |
| CN105112349B (en) | A kind of brucella melitensis molecular marker vaccine strain and its application | |
| KR102165358B1 (en) | Vaccine composition for foot and mouth disease virus a serotype | |
| CN103908665B (en) | A kind of vaccine combination and its preparation method and application | |
| CN104087559B (en) | A kind of infectious bursa of Fabricius virus, inactivated vaccine and preparation method thereof | |
| CN106146626B (en) | A kind of erysipelothrix ruhsiopathiae subunit vaccine and preparation method and application | |
| CN103705918B (en) | Porcine epidemic diarrhea resisting virus hyper-immune serum and preparation method thereof | |
| CN108251382B (en) | Porcine pseudorabies virus weakening method, porcine pseudorabies virus weakening virus strain, porcine pseudorabies virus vaccine composition and application of porcine pseudorabies virus weakening virus strain | |
| CN102727883A (en) | Combined live vaccine against porcine reproductive and respiratory syndrome and swine fever, and application thereof | |
| CN106995489B (en) | A kind of Streptococcus suis truncated protein Sao and application | |
| CN108410784B (en) | Streptococcus suis delta CPS/SsnA-mSly (P353L) -SC19 engineering bacteria and application thereof in vaccines | |
| CN104248754B (en) | Streptococcus suis vaccine combination and its preparation method and application |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |