CN102166348B - 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
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- CN102166348B CN102166348B CN 201010618516 CN201010618516A CN102166348B CN 102166348 B CN102166348 B CN 102166348B CN 201010618516 CN201010618516 CN 201010618516 CN 201010618516 A CN201010618516 A CN 201010618516A CN 102166348 B CN102166348 B CN 102166348B
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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 for the preparation of 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 intensive and industrialized development process, because the application of Biotechnology in Genetic Breeding, feed formulation technology, 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.Major embodiment in process of production since a variety of causes particularly summer fever stress and often cause the animal low or weary feelings of performance of oestrusing, conception rate is low or return the 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 livestocks.Wherein, for sow, the prior art great majority are to adopt the promoting sexual gland hormone (such 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 from the negative feedback Auto-regulator (such as inhibin and follistatin) of the body of dominant follicle and secondary follicle itself, and inhibin can suppress by negative feedback mechanism the secretion of hypophysis FSH, 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 that Anestrus sows is promoted the sexual maturity, return the high problem of feelings rate although often have the final performance that is difficult to be impregnated of quite a few pig rutting.
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 not relating to it is promoting the sexual maturity to Anestrus sows and is promoting application aspect the breeding.
Summary of the invention
One object of the present invention is to overcome the deficiency that prior art is promoted the sexual maturity to Anestrus sows and promoted the existence of breeding aspect, for a variety of causes particularly summer fever stress cause oestrus performance low or weary feelings, conception rate of weary feelings sow at advanced age and weary feelings replacement gilt low or return the problem such as feelings rate rising, 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 the 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 commonly use prepares also can be applied to the present invention.Prepare respectively oil phase, water 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, the 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;
The volume ratio of (2) 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 in preparation, described inhibin recombination fusion protein is applied to prepare inhibin genetic engineering regulation antigen or other drug.
The invention provides preferred inhibin genetic engineering regulation antigen application scheme:
For the first time immunity contains 0.5mg～1mg inhibin recombination fusion protein oil emulsion vaccine for 1ml, or can proceed for the second time immunity in rear 15～21 days in for the first time immunity, for the second time immunity contains 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 within 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 for the third time booster immunization at the 42nd day.General sow normal rutting after twice immunity.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 well litter size of pig.
(3) the inventive method is simple, easily operation, and time advance two all vaccinate that general as required sow performance is oestrused 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 again to oestrus and normally breeding, and improve well number born of sow and reproductive performance, 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 consist of under the heat stress state, can effectively promote oestrus of sow, successfully breeding and normally be impregnated and improve litter size, 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 impact 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 represents the immunosuppressant group, and triangle represents matched group;
The impact 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 represents the immunosuppressant group, and triangle represents 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 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 delivering among the GenBank (Genbank number: NM_214189), design 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 gene coding region sequence.
B, get blue pool pig ovary tissue, place 1ml guanidinium isothiocyanate phenol solution (Trizol liquid, be purchased from Invitrogen company) middle homogenate, then remove protein with chloroform, take out supernatant after centrifugal 15 minutes, add the again total geneome RNA of centrifugation of isopropyl alcohol 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, the pig inhibin code cDNA fragment that amplification is obtained are connected with cloned plasmids PMD18-T carrier (be purchased from Shanghai and give birth to worker company), and the recombinant plasmid transformed that then structure is obtained enters that escherichia coli DH 5 strains (being purchased from Invitrogen company) copy, persistence 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 (being purchased from Invitrogen company):
A, according to the nucleotide sequence of the resulting pig inhibin cDNA of order-checking, by the cDNA sequence of its mature peptide, the synthetic forward primer Mf of design and downstream primer Mr, wherein,
M
f（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 the 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, under the catalysis of Pfu archaeal dna polymerase, again amplify length as 423bp, two ends cDNA cloned sequence with the pig inhibin mature peptide of Bgl II and EcoR I restriction enzyme site through the PCR reaction as template and dNTP take pig inhibin coding region cDNA fragment, be used for inserting expression plasmid pRSET A(and be purchased from Invitrogen company).
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 combination can occur 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, the solution that then will contain this albumen flush away carbamide wherein of dialysing one by one.
(4) prepare respectively oil phase, water 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 its last childbirth occurs in summer, because summer high temperature weather causes long-time weary feelings (average 27 days nonpregnant phases), be divided at random 2 groups after the wean, every group 9, be called I group and C group.Wherein I organizes sow immunosuppressant recombination fusion protein oil emulsion vaccine, and C organizes at one time injecting normal saline adjuvant.
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 Recombinant Swine inhibin fusion rotein, for the second time immunity uses 1mL to contain the oil emulsion vaccine of 0.25mg Recombinant Swine 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, only have 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 immune IHN group average litter size is 12.25 ± 1.03, compares significant difference with 9.3 ± 0.67 of matched group.Show the rate of oestrusing and the litter size that can effectively improve sow to the sow immunosuppressant that is no lack of feelings.
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
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 Recombinant Swine inhibin fusion rotein to A group replacement gilt for the first time, for the second time immunity 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 blood after the immunity:
The method that adopts conventional ELISA method to detect antibody titer detects the effect of the immunity of active immunity inhibin.Utilize the inhibin recombiant protein to be coated with 96 hole ELISA Plate (every hole 100 mL contain 6 mg albumen).The backward every hole of sealase target adds plasma sample (blood plasma is from experiment sow blood) 100 mL of 1600 times of dilutions, then hatches 1h so that anti-inhibin antibody and the inhibin protein binding that is coated with in 37 ℃.The anti-pig IgG of rabbit that then uses horseradish peroxidase-labeled, uses to add volume by volume concentration and be 0.03%H in conjunction with the anti-IHN antibody that is derived from the blood as second antibody (Santa Cruz Biotechnology, SantaCruz, CA, USA) at last
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 (after the wean above 1 month) non-estrus, most of sow (near 80%) is oestrusing about two weeks after the immunity, and remaining shows after for the second time immunity oestruses and breed.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 widely 33.3% of matched group.
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 in addition, about 60% sow performance is oestrused and is bred.Matched group then only has 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 (3)
1. the application of an inhibin recombination fusion protein is characterized in that being applied to preparing and promotes sow and replacement gilt is oestrused, successfully breeding and vaccine or inhibin genetic engineering regulation antigen or the medicine aspect of normally being impregnated at advanced age;
Described application is to adopt the method for active immunity described inhibin genetic engineering regulation antigen to be applied to the breeding endocrine of pig;
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, the 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 it is characterized in that adopting the method for active immunity described inhibin genetic engineering regulation antigen to be applied to the breeding endocrine of pig; The method of described immunity is to contain 0.5mg～1mg inhibin recombination fusion protein oil emulsion vaccine at 1ml for for the first time immunity.
3. application according to claim 2, 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 for the first time immunity, carried out the immunity second time in 15～21 days after for the first time immunity, for the second time immunity contains 0.25mg～0.5 mg inhibin recombination fusion protein oil emulsion vaccine for 1ml.
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| 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 |
| 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 |
| AU2022416227A1 (en) * | 2021-12-15 | 2024-01-04 | Ningbo Sansheng Biological Technology Co., Ltd. | Antibody against inhibin and use thereof |
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| CN101423833A (en) * | 2008-10-21 | 2009-05-06 | 华南农业大学 | Method for preparing inhibin genetic engineering regulation antigen |
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Non-Patent Citations (5)
| Title |
|---|
| B. F. King et al.Ovulatory and endocrine responses after active immunization of gilts against a synthetic fragment of bovine inhibin.《Journal of Animal Science》.1993,第71卷975-982. * |
| Fertility》.1990,第90卷199-205. * |
| R. W. Brown et al.Immunization against recombinant bovine inhibin α subunit causes increased ovulation rates in gilts.《Journal of Reproduction & Fertility》.1990,第90卷199-205. |
| R. W. Brown et al.Immunization against recombinant bovine inhibin α subunit causes increased ovulation rates in gilts.《Journal of Reproduction & * |
| 彭志兰等.抑制素免疫在动物繁殖中应用的研究进展.《中国畜牧兽医》.2007,第34卷(第2期),69-73. * |
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