CN102166348A - Application of inhibin recombinant fusion protein to preparing medicines for promoting oestrus and hybridization of sows - Google Patents
Application of inhibin recombinant fusion protein to preparing medicines for promoting oestrus and hybridization of sows Download PDFInfo
- Publication number
- CN102166348A CN102166348A CN 201010618516 CN201010618516A CN102166348A CN 102166348 A CN102166348 A CN 102166348A CN 201010618516 CN201010618516 CN 201010618516 CN 201010618516 A CN201010618516 A CN 201010618516A CN 102166348 A CN102166348 A CN 102166348A
- Authority
- CN
- China
- Prior art keywords
- inhibin
- fusion protein
- immunity
- sows
- breeding
- 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
Images
Abstract
The invention discloses an application of an inhibin recombinant fusion protein to preparing medicines for promoting oestrus and hybridization of sows. When inhibin is used for the sows: 0.5-1mg of the inhibin recombinant protein is used for initial immunization; and after 15-21 days, 0.25-0.5mg of the inhibin recombinant protein is used for second immunization. The immune inhibin can increase the estrus rate and the litter sizes of the sows. The invention has obvious effects on overcoming anestrus of the sows, caused by heat stress in summer, promoting the breeding activities of breeding sows, and particularly on promoting oestrus of breeding sows and back-up sows and increasing litter sizes. Thus, the non-productive time of the breeding sows is shortened, the breeding performance of the breeding sows is improved, and the economic benefit of sow raising is increased.
Description
Technical field
The invention belongs to the immunoregulation new technical field of regulation and control pig breeding activity, be specifically related to a kind of inhibin recombination fusion protein preparation be used to overcome summer fever stress harmful effect, promote the application of the medicine aspects, aspect such as performance and breeding of oestrusing of sow.
Background technology
Present animal husbandry is in intensification and industrialized development process, because the application of genetic breeding technology, feedstuff compounding technique, epidemic prevention and control technology, environmental health technology and modern enterprise technique of management and administration is greatly improved the production efficiency of animal husbandry.But in each production link, the reproductive performance of animal does not but improve in the past decades.Be mainly reflected in the production process since a variety of causes particularly summer fever stress and often cause the animal low or weary feelings of performance of oestrusing, the breeding conception rate is low or return phenomenon such as feelings rate rising.
In order to improve the reproductive performance of animal, a lot of methods are attempted and inquired into to the researcher of a lot of herding industries.Wherein, for sow, the prior art great majority are to adopt the promoting sexual gland hormone (as FSH) of injection external source to promote the animal follicular development and promote the oestrus of sow breeding.This way tends to excite a large amount of generations of regulating material (as inhibin and follistatin) from the negative feedback of the body of dominant follicle and secondary follicle itself, and inhibin can suppress the secretion of hypophysis FSH by negative feedback mechanism, the follicle that also directly suppresses growth, the quality of reduction follicle and ovum.Follistatin also can reduce the quality of follicle ovum in the growth by suppressing its biological function in conjunction with activin (ACT) neutralization.Therefore when using other gonadotrophins medicines, return the high problem of feelings rate though often have the breeding final performance that is difficult to be impregnated of oestrusing of quite a few pig to weary feelings sow with functions of promoting sexual maturity.
The applicant provides the preparation method of a kind of inhibin recombination fusion protein and regulation antigen in for 200810218477.1 patent application at application number, and the inhibin recombination fusion protein and the regulation antigen that propose to obtain can be applied to improve milch cow application aspect available embryo's number in embryo transfer.But do not relate to it in the application aspect weary feelings sow with functions of promoting sexual maturity and promotion breeding.
Summary of the invention
One object of the present invention is to overcome the deficiency of prior art to weary feelings sow with functions of promoting sexual maturity and promotion breeding aspect existence, at a variety of causes particularly summer fever stress cause weary feelings sow at advanced age and weary feelings replacement gilt oestrus the low or weary feelings of performance, the breeding conception rate is low or return problem such as feelings rate rising, and a kind of new application of inhibin recombination fusion protein is provided.
A further object of the invention provides described inhibin recombination fusion protein and promotes oestrus of sow, successfully breeding and vaccine of normally being impregnated or the application aspect the medicine in preparation.
Above-mentioned purpose of the present invention is achieved by the following technical programs:
The preparation method of inhibin recombination fusion protein of the present invention is described in detail in the applicant's application number is 200810218477.1 patent application, simultaneously, according to the applicant's invention technology, the inhibin recombination fusion protein that the eucaryon method that adopts those skilled in the art to use always prepares also can be applied to the present invention.Prepare oil phase, water respectively according to following steps, oil phase and water be mixed in proportion obtain described immunogen:
(1) be that the inhibin recombination fusion protein (detailed can be description in 200810218477.1 referring to number of patent application) of 2～6 mg/mL is dissolved in the aqueous solution that contains 2～4% Tween 80 and makes water with concentration;
The preparation method of described inhibin recombination fusion protein may further comprise the steps:
(a) according to existing inhibin gene sequence, design specificity forward primer I1 and downstream primer I2;
Described forward primer I1 and downstream primer I2 nucleotides sequence are classified as:
I1：5’-ATGTGGCCTCAGCTGCTCCTCTTGC-3’；
I2：5’-TTAGATGCAGGCACAGTGCTGGGTG-3’；
(b) take guanidinium isothiocyanate-chloroform-isopropyl alcohol method from the pig ovary tissue of the blue pool, to extract total geneome RNA, obtain cDNA by reverse transcription;
(c) synthesize cDNA as template with primer I 1, I2, dNTP and reverse transcription, under the catalysis of archaeal dna polymerase, react the code cDNA fragment of amplification pig inhibin by PCR;
(d) according to the cDNA base sequence of inhibin mature peptide and the multiple clone site base sequence of colibacillus expression plasmid pRSET A, synthetic forward primer Mf of design and downstream primer Mr;
Described forward primer Mf and downstream primer Mr nucleotides sequence are classified as:
Mf：5’-AATAGATCTTCTCCACCGCCCCTCTGCCC-3’；
Mr：5’-ATCGAATTCAGATGCAGGCACAGTGCTGGG-3’；
(e) utilize the coded cDNA sequence of the inhibin that described primer Mf, Mr and above-mentioned post transcription cloning obtain to be template, amplify the cDNA cloned sequence of inhibin mature peptide by the PCR reaction;
(f) behind the cDNA cloned sequence of described inhibin mature peptide process Bgl II and the EcoR I double digestion, be cloned between the Bgl II and EcoR I two restriction enzyme sites of expression plasmid pRSET A, be built into recombinant expression plasmid pInhibin SCAU;
(g) described recombinant expression plasmid pInhibin SCAU is transformed into e. coli strains, the IPTG that adds 0.05mmol/L～0.2 mmol/L in culture fluid induces, and obtains the inhibin recombination fusion protein that recombinant bacteria is expressed;
(2) volume ratio of No. 10 lightweight white oils and Si Ben 80 being pressed 94%:6% is mixed, and adds concentration (w:v) again and be 2% aluminium stearate, and mix homogeneously is made oil phase;
(3) water and oil phase are mixed by 4～5: 5～6 volume ratio, and stir and make water in oil homogenizing emulsion, i.e. inhibin genetic engineering regulation antigen.
The invention provides described inhibin recombination fusion protein and promote oestrus of sow, successfully breeding and normally be impregnated and improve application aspect the litter size vaccine, described inhibin recombination fusion protein is applied to prepare inhibin genetic engineering regulation antigen or other drug in preparation.
The invention provides preferred inhibin genetic engineering regulation antigen application scheme:
Immunity for the first time contains 0.5mg～1mg inhibin recombination fusion protein oil emulsion vaccine for 1ml, or can proceed immunity for the second time in 15～21 days in the immunity back first time, the second time, immunity contained 0.25mg～0.5 mg inhibin recombination fusion protein oil emulsion vaccine for 1ml.
The present invention adopts the approach of " active immunity " described inhibin recombination fusion protein to be applied to the breeding endocrine of pig, immune programme for children is: the 1st day this recombiant protein of first immunisation, do not oestrus if experiment pig did not show in 3 weeks, then carried out booster immunization at the 21st day.Behind the booster immunization, if experiment pig after 3 weeks in non-estrus still, then carried out booster immunization for the third time at the 42nd day.The breeding of after twice immunity, can normally oestrusing of general sow.Adopt the approach of intramuscular injection to carry out the injection of vaccine.
The present invention compared with prior art has following beneficial effect:
(1) inhibin genetic engineering regulation antigen avirulence, no pathogenicity, the drug residue free of the present invention's preparation to the person poultry harmless, are successfully realized effect of the present invention on the basis that the consumer to edible Carnis Sus domestica or its goods has no adverse effects.
(2) creatively improve through of the present invention, with immunosuppressant vaccine aspect be applied to promote oestrus of sow, successfully the breeding and normally be impregnated, can promote more that than additive method pig oestruses, reduces nonproductive time, and can improve litter size of pig well.
(3) the inventive method is simple, operation easily, and the time that the general performance of sow is as required oestrused is put forward the last fortnight vaccinate, can produce extraordinary effect.
Experimental result of the present invention shows, utilize the immunosuppressant of the technology of the present invention preparation, can promote the sow of long-term non-estrus to oestrus again and normally breeding, and improve number born of sow and reproductive performance well, this can solve oestrusing of pig farm and postpone or the non-estrus problem, reduces the nonproductive time of pig farm sow.The present invention has widened the range of application of inhibin, the application of exploitation immunosuppressant technology aspect raising broad sow reproductive performance.By to being in the Emulsion immunogen that weary feelings sow at advanced age and weary feelings replacement gilt immunosuppressant recombiant protein constitute under the heat stress state, can promote oestrus of sow, successfully breeding and normally be impregnated and improve litter size effectively, the efficient that promotes pig industry to produce is all had good meaning and application prospect with increasing economic efficiency.
Description of drawings
Fig. 1 active immunity inhibin (■) and the anti-inhibin antibody titer of matched group sow () blood in experiment, data are average ± standard error, on the same group the different alphabetical person's significant differences (a, b:P＜0.05) of data subscript;
The electrophoresis pattern of Fig. 2 inhibin gene coding region;
The influence that Fig. 3 active immunity inhibin changes weary feelings oestrus of sow at advanced age, arrow representative injection inhibin and blood-sample withdrawal time, round dot is represented the immunosuppressant group, and triangle is represented matched group;
The influence that Fig. 4 active immunity inhibin is oestrused and changed weary feelings replacement gilt, arrow representative injection inhibin and blood-sample withdrawal time, round dot is represented the immunosuppressant group, and triangle is represented matched group.
The specific embodiment
Further describe the present invention below in conjunction with embodiment, employed test method is conventional method if no special instructions among the following embodiment; Employed material, reagent etc. if no special instructions, are the reagent and the material that can obtain from commercial channels.
Embodiment 1The preparation of pig immunosuppressant vaccine
(1) clone of pig inhibin encoding gene cDNA:
A, according to the pig inhibin gene sequence of being delivered among the GenBank (Genbank number: NM_214189), design the specificity forward primer I1 and the downstream primer I2 of this gene.Forward primer I1(5 '-ATGTGGCCTCAGCTGCTCCTCTTGC-3 ') corresponding to the 1st～25 nucleotide in pig inhibin gene coding region; Downstream primer I2(5 '-TTAGATGCAGGCACAGTGCTGGGTG-3 ') corresponding to the 1071st～1095 nucleotide in pig inhibin gene coding region.Described pig inhibin gene total length 1095bp contains complete inhibin α subunit gene coding region sequence.
B, get blue pool pig ovary tissue, place 1ml guanidinium isothiocyanate phenol solution (Trizol liquid, purchase company in Invitrogen) in homogenate, remove protein with chloroform then, take out supernatant after centrifugal 15 minutes, add the isopropyl alcohol total geneome RNA of centrifugation again in supernatant, the concentration that total RNA is made into 1g/L is dissolved in the water that contains DEPC, and (RT) obtains cDNA by reverse transcription.
C, usefulness primer I 1, I2, dNTP and the synthetic cDNA of reverse transcription are as template, under the catalysis of archaeal dna polymerase by the PCR reaction amplification specific band that to obtain a length be 1095bp, be the code cDNA fragment of pig inhibin, the electrophoresis pattern of the inhibin gene coding region behind the post transcription cloning as shown in Figure 2.
D, pig inhibin code cDNA fragment that amplification is obtained and cloned plasmids PMD18-T carrier (purchase give birth in Shanghai worker company) are connected, then the recombinant plasmid transformed that structure is obtained go into that escherichia coli DH 5 strains (purchasing the company in Invitrogen) are duplicated, preservation forever and carry out nucleotide sequencing.SEQ ID NO:1 is the nucleotide sequence of the pig inhibin cDNA that obtains with the method post transcription cloning in the sequence table, and wherein capitalization part is distinguished corresponding I1 and I2 primer; This consecutive nucleotides is the coded cDNA sequence of pig inhibin mature peptide, and therefore the pig inhibin gene nucleotide sequence of this amplification and the homology of other domestic animal can prepare inhibin genetic engineering regulation antigen with pig inhibin code cDNA fully more than 90%.
(2) recombination fusion protein of preparation pig inhibin: utilize the cDNA base sequence of pig inhibin mature peptide, can carry out the expression of pig inhibin recombination fusion protein by prokaryotic expression system E.coli BL21 (DE3) strain (purchasing company) in Invitrogen:
A, according to the nucleotide sequence of the resulting pig inhibin cDNA of order-checking, by the cDNA sequence of its mature peptide, synthetic forward primer Mf of design and downstream primer Mr, wherein,
Mf（5’-AATAGATCT
TCTCCACCGCCCCTCTGCCC-3’）
Mf is corresponding to the 691st～708 nucleotide sequence of the coding region of sequence pig inhibin gene shown in the SEQ ID NO:1, and square frame partly contains Bgl II restriction enzyme site, and setting-out partly is a base of keeping reading frame;
Mr（5’-ATCGAATTCAGATGCAGGCACAGTGCTGGG-3）
Mr is corresponding to the 1073rd～1093 nucleotide sequence of the coding region of sequence pig inhibin gene shown in the SEQ ID NO:1, and square frame partly contains EcoR I restriction enzyme site.
B, utilize primer Mf, Mr, once more amplifying length through the PCR reaction as template and dNTP with pig inhibin coding region cDNA fragment under the catalysis of Pfu archaeal dna polymerase is the cDNA cloned sequence that 423bp, two ends have the pig inhibin mature peptide of Bgl II and EcoR I restriction enzyme site, is used to insert expression plasmid pRSET A(and purchases the company in Invitrogen).
C, with this recombinant expression plasmid pInhibin SCAU and Transformed E coli.BL21 (DE3) strain, the IPTG that adds 0.05mmol/L～0.2mmol/L in culture fluid induces and expressed pig inhibin recombination fusion protein in 4～6 hours, expression is expressed inhibin a subunit recombination fusion protein near the band of 20kD in the swimming lane 2 wherein as shown in Figure 4.This pig inhibin recombination fusion protein can be by the anti-cattle a-of rabbit inhibin antibody specific recognition.
(3) purification of pig inhibin recombination fusion protein:
Utilize the aminoterminal of the pig inhibin recombination fusion protein of escherichia coli expression to have the labelling (His-tag) of 6 continuous histidine, special affine the combination can take place with nickel ion in this His-Tag labelling.Therefore, adopt the cracking of 8M urea liquid to express the antibacterial of pig inhibin recombination fusion protein, with lysate process Ni-NTA resin gel chromatography, can be purified into wherein pig inhibin recombination fusion protein and with recombiant protein, and this albumen also can be by the anti-cattle of rabbit-inhibin antibody specific recognition, will contain this proteic solution flush away carbamide wherein of dialysing one by one then.
(4) prepare oil phase, water respectively according to following ratio, oil phase and water are mixed in proportion obtain described pig immunosuppressant vaccine (immunogen);
(a) be that the above-mentioned expression, purification of 2～6 mg/mL and the pig inhibin recombination fusion protein after the dialysis (can be 200810218477.1 description) be dissolved in the aqueous solution that contains 2～4% Tween 80 and make water with concentration referring to number of patent application;
(b) No. 10 lightweight white oils and Si Ben 80 are mixed by 94% and 6% volume ratio, add concentration (w:v) again and be 2% aluminium stearate, mix homogeneously is made oil phase;
(c) water and oil phase are mixed by 4～5: 5～6 volume ratio, and stir and make water in oil homogenizing emulsion, i.e. inhibin genetic engineering regulation antigen.
Select 18 sows at advanced age that produced 4 or 5 tires (average parity is the 4.7th tire), because of its last childbirth occurs in summer, the wean back is divided into 2 groups at random because of summer high temperature weather causes weary for a long time feelings (average 27 days nonpregnant phases), every group 9, be called I group and C group.Wherein I organizes sow immunosuppressant recombination fusion protein oil emulsion vaccine, and C organizes injecting normal saline adjuvant at one time.
Immune programme for children:
Sow to the I group is injected an inhibin recombination fusion protein oil emulsion vaccine every 21 d, use for the first time 1mL to include the oil emulsion vaccine of 0.5mg reorganization pig inhibin fusion rotein, immunity for the second time uses 1mL to contain the oil emulsion vaccine of 0.25mg reorganization pig inhibin fusion rotein.
Result of the test shown in the table 1 shows: 9 sows of immunosuppressant all oestrused in 40 days, and the accumulative total rate of oestrusing is 100%; And during to 100 days by a definite date off-test, have only 3 hair feelings in 9 pigs of matched group, the rate of oestrusing is 33.3%, sees shown in the accompanying drawing 3.
The number born of sow of oestrusing is carried out tracing record and statistics, and it is 12.25 ± 1.03 that immune IHN organizes average litter size, compares significant difference with 9.3 ± 0.67 of matched group.Show the rate of oestrusing and the litter size that can improve sow to the sow immunosuppressant that is no lack of feelings effectively.
Table 1: the accumulative total of active immunity inhibin and matched group pig oestrus rate and litter size.
| Group | n | The sow number of oestrusing | The rate of oestrusing (%) | Average litter size |
| Experimental group | 9 | 9 | 100 | 12.25±1.03 b |
| Matched group | 9 | 3 | 33.3 | 9.30±0.67 a |
Annotate: with the different alphabetical person's significant differences (a, b:P＜0.05) of column data subscript
Embodiment 3
20 of the replacement gilts of selecting for 8 monthly ages and not oestrusing are divided into 2 groups at random, 10 every group, are called A group and B group.Wherein to A group replacement gilt immunosuppressant recombination fusion protein oil emulsion vaccine, B group replacement gilt is the injecting normal saline adjuvant at one time.
Immune programme for children:
Every inhibin recombination fusion protein oil emulsion vaccine of 21d injection, use 1mL to contain the vaccine of 0.5mg reorganization pig inhibin fusion rotein to A group replacement gilt for the first time, immunity for the second time uses 1mL to contain the vaccine of 0.25mg inhibin recombination fusion protein.
Experimental result shows: accumulative total the oestrus rate of 10 replacement gilts in 50 days of immunosuppressant is 60%, and matched group only is 30%, sees shown in the accompanying drawing 3.
Embodiment 4Index detects
(1) the anti-inhibin antibody titer of immunity back blood:
The method that adopts conventional ELISA method to detect antibody titer detects the effect of the immunity of active immunity inhibin.Utilize inhibin recombiant protein bag by 96 hole ELISA Plate (every hole 100 mL contain 6 mg albumen).Add plasma sample (blood plasma is from experiment sow blood) 100 mL of 1600 times of dilutions behind the sealase target to every hole, hatch 1h so that anti-inhibin antibody and the inhibin protein binding of wrapping quilt in 37 ℃ then.(CA USA) in conjunction with the anti-IHN antibody that is derived from the blood, uses to add volume by volume concentration and be 0.03%H the anti-pig IgG of rabbit that then uses horseradish peroxidase-labeled at last for Santa Cruz Biotechnology, SantaCruz as second antibody
2O
2TMB solution colour developing, after with 1mol/L sulphuric acid cessation reaction, measure the absorbance value of 450 nm wavelength as antibody titer with microplate reader (Bio-Rad).
After the immunity, anti-inhibin antibody titer will be significantly higher than non-immune matched group pig in the sow blood, sees shown in the accompanying drawing 1.
(2) oestrusing and litter size of sow:
Behind multiparity sow injection at the advanced age inhibin vaccine to long-term (the wean back was above 1 month) non-estrus, most of sow (near 80%) is oestrusing about two weeks after the immunity, and remaining is oestrused and breed in immunity back performance for the second time.And concurrent control group pig only about 30% performances oestrus and breed, see shown in the accompanying drawing 3.Result during off-test shows: the oestrus of sow rate of immunosuppressant group is 100%, is higher than 33.3% of matched group widely.
Through a trimester of pregnancy, immune Farrowing is on average farrowed 12.25 ± 1.03, and remarkable (P＜0.05) is higher than 9.30 ± 0.67 of matched group.Behind the replacement gilt immunosuppressant of not oestrusing to reaching for 8 monthly ages, about 60% sow performance is oestrused and is bred in addition.Matched group then has only 30% performance to oestrus at 50 days duration of test, sees shown in the accompanying drawing 4.
SEQUENCE?LISTING
＜110〉Agricultural University Of South China
＜120〉application of inhibin recombination fusion protein aspect preparation promotion oestrus of sow and breeding medicine
<130>
<160> 4
<170> PatentIn?version?3.3
<210> 1
<211> 25
<212> DNA
＜213〉primer I 1
<400> 1
atgtggcctc?agctgctcct?cttgc 25
<210> 2
<211> 25
<212> DNA
＜213〉primer I 2
<400> 2
ttagatgcag?gcacagtgct?gggtg 25
<210> 3
<211> 29
<212> DNA
＜213〉primer Mf
<400> 3
aatagatctt?ctccaccgcc?cctctgccc 29
<210> 4
<211> 30
<212> DNA
＜213〉primer Mr
<400> 4
atcgaattca?gatgcaggca?cagtgctggg 30
Claims (5)
1. the application of an inhibin recombination fusion protein is characterized in that being applied to preparation and promotes oestrus of sow, successfully breeding and normally be impregnated vaccine or inhibin genetic engineering regulation antigen or medicine aspect;
The preparation method of described inhibin recombination fusion protein may further comprise the steps:
(a) according to existing inhibin gene sequence, design specificity forward primer I1 and downstream primer I2;
Described forward primer I1 and downstream primer I2 nucleotides sequence are classified as:
I1：5’-ATGTGGCCTCAGCTGCTCCTCTTGC-3’；
I2：5’-TTAGATGCAGGCACAGTGCTGGGTG-3’；
(b) take guanidinium isothiocyanate-chloroform-isopropyl alcohol method from the pig ovary tissue of the blue pool, to extract total geneome RNA, obtain cDNA by reverse transcription;
(c) synthesize cDNA as template with primer I 1, I2, dNTP and reverse transcription, under the catalysis of archaeal dna polymerase, react the code cDNA fragment of amplification pig inhibin by PCR;
(d) according to the cDNA base sequence of inhibin mature peptide and the multiple clone site base sequence of colibacillus expression plasmid pRSET A, synthetic forward primer Mf of design and downstream primer Mr;
Described forward primer Mf and downstream primer Mr nucleotides sequence are classified as:
Mf：5’-AATAGATCTTCTCCACCGCCCCTCTGCCC-3’；
Mr：5’-ATCGAATTCAGATGCAGGCACAGTGCTGGG-3’；
(e) utilize the coded cDNA sequence of the inhibin that described primer Mf, Mr and above-mentioned post transcription cloning obtain to be template, amplify the cDNA cloned sequence of inhibin mature peptide by the PCR reaction;
(f) behind the cDNA cloned sequence of described inhibin mature peptide process Bgl II and the EcoR I double digestion, be cloned between the Bgl II and EcoR I two restriction enzyme sites of expression plasmid pRSET A, be built into recombinant expression plasmid pInhibin SCAU;
(g) described recombinant expression plasmid pInhibin SCAU is transformed into e. coli strains, the IPTG that adds 0.05mmol/L～0.2 mmol/L in culture fluid induces, and obtains the inhibin recombination fusion protein that recombinant bacteria is expressed.
2. application according to claim 1 is characterized in that being applied to preparing and promotes oestrus of sow, the successfully breeding or the inhibin genetic engineering regulation antigen of normally being impregnated;
The preparation method of described inhibin genetic engineering regulation antigen may further comprise the steps:
(h) be that the inhibin recombination fusion protein of 2～6 mg/mL is dissolved in the aqueous solution that contains 2～4% tween 80s and makes water with concentration;
(i) No. 10 lightweight white oils and Si Ben 80 are mixed, add aluminium stearate again, mix homogeneously is made oil phase;
(j) water and oil phase are mixed, and stir and make water in oil homogenizing emulsion, promptly get inhibin genetic engineering regulation antigen.
3. application according to claim 2 is characterized in that it being to adopt the method for active immunity described inhibin genetic engineering regulation antigen to be applied to the breeding endocrine of pig.
4. application according to claim 3 is characterized in that the method for immunity is to contain 0.5mg～1mg inhibin recombination fusion protein oil emulsion vaccine at 1ml for immunity for the first time.
5. application according to claim 4, the method that it is characterized in that immunity is to contain 0.5mg～1mg inhibin recombination fusion protein oil emulsion vaccine for 1ml for immunity for the first time, carried out the immunity second time in 15～21 days after immunity for the first time, immunity for the second time contains 0.25mg～0.5 mg inhibin recombination fusion protein oil emulsion vaccine for 1ml.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010618516 CN102166348B (en) | 2010-12-31 | 2010-12-31 | Application of inhibin recombinant fusion protein to preparing medicines for promoting oestrus and hybridization of sows |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010618516 CN102166348B (en) | 2010-12-31 | 2010-12-31 | Application of inhibin recombinant fusion protein to preparing medicines for promoting oestrus and hybridization of sows |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102166348A true CN102166348A (en) | 2011-08-31 |
| CN102166348B CN102166348B (en) | 2013-01-09 |
Family
ID=44487790
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201010618516 Expired - Fee Related CN102166348B (en) | 2010-12-31 | 2010-12-31 | Application of inhibin recombinant fusion protein to preparing medicines for promoting oestrus and hybridization of sows |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102166348B (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103041381A (en) * | 2013-01-09 | 2013-04-17 | 江苏省农业科学院 | Recombinant inhibin vaccine and preparation method and application of recombinant inhibin vaccine |
| CN104012466A (en) * | 2014-06-10 | 2014-09-03 | 江苏省农业科学院 | Method for promoting oestrus of cattle and increasing hybridization conception rate of cattle |
| CN111134084A (en) * | 2020-01-06 | 2020-05-12 | 浙江省农业科学院 | Method for optimizing timing insemination effect of replacement gilts |
| WO2023109803A1 (en) * | 2021-12-15 | 2023-06-22 | 宁波三生生物科技股份有限公司 | Antibody against inhibin and use thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101423833A (en) * | 2008-10-21 | 2009-05-06 | 华南农业大学 | Method for preparing inhibin genetic engineering regulation antigen |
-
2010
- 2010-12-31 CN CN 201010618516 patent/CN102166348B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101423833A (en) * | 2008-10-21 | 2009-05-06 | 华南农业大学 | Method for preparing inhibin genetic engineering regulation antigen |
Non-Patent Citations (3)
| Title |
|---|
| 《Journal of Animal Science》 19931231 B. F. King et al Ovulatory and endocrine responses after active immunization of gilts against a synthetic fragment of bovine inhibin 975-982 第71卷, * |
| 《Journal of Reproduction & Fertility》 19901231 R. W. Brown et al Immunization against recombinant bovine inhibin alpha subunit causes increased ovulation rates in gilts 199-205 第90卷, * |
| 《中国畜牧兽医》 20071231 彭志兰等 抑制素免疫在动物繁殖中应用的研究进展 69-73 第34卷, 第2期 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103041381A (en) * | 2013-01-09 | 2013-04-17 | 江苏省农业科学院 | Recombinant inhibin vaccine and preparation method and application of recombinant inhibin vaccine |
| CN104012466A (en) * | 2014-06-10 | 2014-09-03 | 江苏省农业科学院 | Method for promoting oestrus of cattle and increasing hybridization conception rate of cattle |
| CN104012466B (en) * | 2014-06-10 | 2016-01-20 | 江苏省农业科学院 | Promote that ox oestruses and improves the method for ox conception rate |
| CN111134084A (en) * | 2020-01-06 | 2020-05-12 | 浙江省农业科学院 | Method for optimizing timing insemination effect of replacement gilts |
| WO2023109803A1 (en) * | 2021-12-15 | 2023-06-22 | 宁波三生生物科技股份有限公司 | Antibody against inhibin and use thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102166348B (en) | 2013-01-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Gifre et al. | Trends in recombinant protein use in animal production | |
| CN102166348B (en) | Application of inhibin recombinant fusion protein to preparing medicines for promoting oestrus and hybridization of sows | |
| Daly et al. | Towards improving the outcomes of assisted reproductive technologies of cattle and sheep, with particular focus on recipient management | |
| TW200846361A (en) | Expression and bioactivity of bovine follicle stimulating hormone | |
| US8367073B2 (en) | Chloramphenicol acetyl transferase (CAT)-defective somatostatin fusion protein and uses thereof | |
| Thakur et al. | Expression and molecular cloning of interferon stimulated genes in buffalo (Bubalus bubalis) | |
| CN110184361B (en) | Molecular marker for identifying genetic sex of hucho taimen and identification method | |
| CN101423833A (en) | Method for preparing inhibin genetic engineering regulation antigen | |
| RU2170100C2 (en) | Method of increase of poultry productivity, fused heterological protein and method of its producing | |
| CN105685503A (en) | Piglet feed additive and preparation method thereof | |
| Reineri et al. | Gene expression of growth factor BMP15, GDF9, FGF2 and their receptors in bovine follicular cells | |
| Kumar et al. | Hypophyseal gene expression profiles of FSH-β, LH-β, and glycoprotein hormone-α subunits in Ictalurus punctatus throughout a reproductive cycle | |
| Huang et al. | Cloning expression and immunogenicity analysis of inhibin gene in Ye Mule Aries sheep | |
| Bhardwaj et al. | Heterologous expression and characterization of indian sahiwal cattle (Bos indicus) alpha inhibin | |
| CN115746146A (en) | GnRH recombinant protein antigen, mammal castration recombinant protein vaccine and preparation method | |
| Said et al. | Hd86 mRNA expression profile in Hyalomma scupense life stages, could it contribute to explain anti-tick vaccine effect discrepancy between adult and immature instars? | |
| Arango G et al. | Association of the bovine growth hormone gene with Holstein cattle reproductive parameters | |
| Rushton | Ovine fascioliasis following reinfection | |
| Roberts et al. | Production of a recombinantly derived growth hormone antibody and the characterization of growth hormone levels in yellow perch | |
| CN110101854B (en) | Carp herpesvirus III type vaccine and preparation method thereof | |
| Naz et al. | Effect of immunization with six sperm peptide vaccines on fertility of female mice. | |
| Olsson et al. | Singular epidural follicle-stimulating hormone administration on follicular growth in camels | |
| CN116375844B (en) | Inhibin antigen peptide, vaccine and preparation method thereof | |
| Schneider et al. | Insulin-like growth factor and growth hormone receptor in postpartum lactating beef cows | |
| Ryskeldina et al. | Obtaining and use of the recombinant bovine pregnancy-associated glycoprotein 1 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20130109 Termination date: 20151231 |
|
| EXPY | Termination of patent right or utility model |