CN103416351A - Method for establishing Bian chicken quick-grow type AA strain and slow-grow type GG strain colony - Google Patents
Method for establishing Bian chicken quick-grow type AA strain and slow-grow type GG strain colony Download PDFInfo
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- CN103416351A CN103416351A CN2013104113822A CN201310411382A CN103416351A CN 103416351 A CN103416351 A CN 103416351A CN 2013104113822 A CN2013104113822 A CN 2013104113822A CN 201310411382 A CN201310411382 A CN 201310411382A CN 103416351 A CN103416351 A CN 103416351A
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Abstract
The invention relates to poultry breeding, in particular to a method for establishing a Bian chicken quick-grow type AA strain and slow-grow type GG strain colony. In a seventh Bian chicken colony of hemp-feather single-comb strain bred in an atresia colony family, the function locus where the myostatin (MSTN) gene affects muscle growing of Bian chickens is determined, the PCR-RFLP technology is used for detecting the genotype of the G2283A locus of the MSTN gene, a male chicken colony and a female chicken colony with the quick-grow genotype (AA) and the slow-grow genotype (GG) are selected for breeding, and the quick-grow type (AA) strain and slow-grow type (GG) strain colony is cultured. The average weights of male chickens and female chickens of 6 years, 8 years, 10 years, 12 years, 14 years and 16 years of the quick-grow type (AA) strain are obviously about 10% higher than those of the male chickens and female chickens of the same period of the slow-grow type (GG) strain.
Description
Technical field
The present invention relates to poultry breeding, be specially a kind of method of setting up the fast elongated AA of limit chicken strain and slow elongated GG strain colony.
Background technology
China's bird genetic resources is the natural resources of local fowl kind preciousness, is also to be worth high economic resources.The genetic diversity of local fowl kind is following poultry breed improvement and adapts to the hereditary basis that the productivity condition changes, and is protection livestock breeding sustainable development and the basic foundation of formulating the reasonable development industrial policy.But, the shortcomings such as local fowl kind is because the level of production is low, and reproduction rate is low, feed consumption is many, early growth is slow, fattening effect is poor and a large amount of external fowl kind, hash unrest is joined, cause the degeneration of local fowl kind quality and the minimizing of quantity, even some good kinds are endangered.Therefore, rationally utilize local fowl kind resource imperative.
At first the animal production modernization is must goodization of kind, therefore planned, on purpose cultivate and improve the breed, improving productivity is animal breeding worker's vital task.In view of animal breeding work has chronicity, the characteristics such as comprehensive, arduous.Animal breeding science is being controlled the animal developing direction, and it is one forces animal to change and the science of development towards the direction that is conducive to mankind's demand, together with it and animal production are closely connected.
Along with the fast development of immunology, electrophoretic techniques and molecular biotechnology, people have also entered a brand-new period to the research of the Biochemical Genetical markers in blood of chicken.Concept to blood type also not only rests on the erythrocyte surface antigen type, but has obtained a brand-new understanding from immunology, electrophoretic techniques.Aspect immunology research, people not only can be according to red cell grouping, and can carry out somatotype according to leucocyte, blood platelet, also can pass through serotype.Aspect Electrophoretic study, can, according to albumen (enzyme) the polymorphism somatotype of serum, also can pass through electropherotyping to cellular content.Aspect the blood type research of chicken, utilize at present immunologic method can separate 14 systems, more than 40 blood factor, aspect the blood Electrophoretic study, albumen (enzyme) polymorphism has been studied more than 30 gene locus.Molecular biological development is people to of genetic material itself, the carrier DNA that is hereditary information has had more understanding, people can carry out the research of RFLP, RAPD and DNA fingerprint aspect now, the research of these aspects has obtained fruitful achievement, having found out QTL site that some and we are concerned about has the genetic marker of genetic correlation, utilize these genetic markers, can indirectly or directly be selected chicken, accelerated the process of breeding work.
Summary of the invention
The object of the present invention is to provide the method for setting up fast elongated (AA) and slow elongated (GG) strain colony.Utilize and determine that muscle growth suppresses the functional site of MSTN effect gene limit chicken muscle growth, for the mark G2283A site of having identified, adopt the PCR-RFLP technology, Select gene type AA, cock, hen that GG is identical, collect seed, hatching, set up fast elongated (AA) and elongated (GG) strain colony slowly.
The present invention adopts following technical scheme to realize:
A kind of method of setting up the fast elongated AA of limit chicken strain and slow elongated GG strain colony, comprise the steps: in the 7th generation numb plumage leaf-comb strain limit chicken group of closed population family breeding, use the PCR-RFLP technology to be detected the genotype in MSTN gene G2283A site, the PCR product is cut with the BbvI enzyme, and the sample called after that will contain 162bp purpose fragment is grown frequency of genotypes AA soon; The sample called after that will contain 127bp purpose fragment is grown genotype GG slowly; Then, choose respectively fast long frequency of genotypes AA and the male and female chicken group breeding of long genotype GG slowly, by the conventional breeding cultural method, turn out the fast elongated AA of limit chicken strain colony and the slow elongated GG of limit chicken strain colony respectively.
The limit chicken belongs to meat egg dual-purpose type kind, and build is medium, is ingot-shaped.Shin is long and sturdy, and crown type be take leaf-comb as main, and a small amount of strawberry hat, pea hat and other crest are arranged.Cock is crown upright, and hen is crown less, and obvious S sigmoid is arranged, and the hat look scarlet.
Line breeding is on the basis that keeps the original productivity of a certain Breeds and body profile basic characteristics, by predeterminated target, carries out directive breeding, creates the breeding mode with special performance strain.Line breeding is indispensable important step in the breeding of new variety process, its important effect that further develops to the formation of new varieties and new varieties.
In the 7th generation numb plumage leaf-comb strain limit chicken group of closed population family breeding, use the PCR-RFLP technology to be detected the genotype in MSTN gene G2283A site, 1 pair of primer of BbvI site design according to chicken MSTN gene order in GenBank (the GenBank accession number is AF346599) for exons 1, the amplified production size is 162 bp.Primer sequence is as follows:
P1：F:?5’-GCATTAGCAGGGACGTTAT-3’;（SEQ?ID?NO：1）
R:5’-ACTCCGTAGGCATTGTGAT-3’?（SEQ?ID?NO：2）
The PCR product is cut with the BbvI enzyme, to detect polymorphism.Restriction enzyme mapping (as shown in Figure 1) shows 3 kinds of genotype, the sample of 162bp purpose fragment is only arranged not containing the BbvI restriction enzyme site, called after AA type; What 127bp was only arranged is to contain the homozygous of BbvI restriction enzyme site, called after GG type; The heterozygous sample for containing the BbvI restriction enzyme site that simultaneously contains 162bp and 127bp, called after GA type.
Choose fast long genotype (AA) and the male and female chicken group breeding of long genotype (GG) slowly, cultivate fast elongated (AA) and elongated (GG) strain colony slowly.The average weight of fast elongated strain (AA) male and female chicken in 6,8,10,12,14,16 week age, the utmost point is significantly higher than slow elongated strain (GG) the body weight same period approximately 10%.
The present invention is reasonable in design, utilizes the numb plumage leaf-comb strain of limit chicken closed population family breeding, by molecular mark, has set up fast elongated AA and slow elongated GG strain colony.
The accompanying drawing explanation
Fig. 1 is the restriction enzyme mapping schematic diagram.
Fig. 2 is that schematic diagram is bred by limit chicken AA, GG colony.
Embodiment
Below the specific embodiment of the present invention is elaborated.
A kind of method of setting up the fast elongated AA of limit chicken strain and slow elongated GG strain colony, comprise the steps:
(1), the 7th from generation to generation in numb plumage leaf-comb strain limit chicken group of closed population family breeding, gather the 7th 299 parts of numb plumage leaf-comb strain hen blood samples, 81 parts of cock blood samples in the chicken core group of limit from generation to generation.According to the detection method (PCR-RFLP) in the chicken MSTN gene G2283A site, limit set up in early stage, the MSTN genotype of institute's blood-sample withdrawal is checked, the PCR product is cut with the BbvI enzyme, and the sample called after that will contain 162bp purpose fragment is grown frequency of genotypes AA soon; The sample called after that will contain 127bp purpose fragment is grown genotype GG slowly, determines AA type and GG type individuality in the chicken fiber crops plumage leaf-comb strain of limit.Determine 75, AA type, 80, GG type, 144, AG type in 299 parts of hen blood samples, in 81 parts of cock blood samples, determine 17, AA type, 13, GG type, 51, AG type.
(2), as shown in Figure 2, in fixed limit chicken fiber crops plumage leaf-comb strain AA type male and female chicken, according to 16 weeks age body weight sorted from big to small, select front 50 hens and front 5 cocks, cohort, gather kind of an egg, (fast length) strain of setting up the AA type; In fixed limit chicken fiber crops plumage leaf-comb strain GG type male and female chicken, according to 16 weeks age body weight sorted from small to large, select front 50 hens and front 5 cocks, cohort, gather kind of an egg, (the growing slowly) strain of setting up the GG type.AA type and GG type homologous genes type cock, hen be cohort respectively, by strain semen deposition, hatching, every chick is dressed to wing number, carry out the subfield raising by strain, raising, management, vaccine program etc. are identical, turn out the fast elongated AA of limit chicken strain colony and the slow elongated GG of limit chicken strain colony.
Daily management will be carried out the raising daily record, comprises day amount of livestock on hand, injures and deaths reason, the feed consumption rate of each strain etc.Enter that incubation is heavy, chick will weigh by only (individual), since 6 weeks age, within every two weeks, by only weighing once, 20 week age, last pursuing only weighed.All Data Collections are subsequently undertaken by limit chicken conservation group requirement.
Need collect seed 500 pieces, egg of each genotype strain, approximately 400 of hatchings, 16 week age, each strain need be butchered 80 of hens, and the residue hen can carry out line breeding by genotype from now on.
Butcher each 50 of the limit chicken hens of AA and GG type 16 week age, measure that chest muscle is heavy, leg flesh is heavy and chest muscle rate, leg flesh rate, relatively the difference between 2 kinds of genotype limit chicken colony testing indexs.Get respectively chest muscle and the leg flesh of the limit chicken hen of 30 AA and GG type and make paraffin section, by omnipotent video imaging system device, examine under a microscope and measure myofibrillar major diameter and minor axis, and calculate the muscle fibre cross-sectional area according to formula: area=major diameter * minor axis * 0.7, the relatively difference of AA and GG type limit chicken muscle fibre cross-sectional area, diameter and density.
Butcher 16 week age AA and GG type limit chicken hen each 30, extract respectively total DNA of chest muscle and leg flesh with kit, transcribed with the reverse transcription kit, across 5 gene primers of intron design, with β-action, as reference gene, each sample arranges 3 repetitions.Adopt real-time fluorescence quantitative PCR (FQ-PCR) to analyze MSTN, MyoD, Myf5, MyoG and the differential expression of Smad3 gene in AA and two colonies of GG, set forth the expression rule between gene.
The experimental record data are as follows:
The average weight in 6,8,10,12,14,16 week age of fast elongated AA strain cock is: 526.48 ± 56.99 grams, 706.42 ± 74.91 grams, 945.07 ± 101.27 grams, 1117.25 ± 126.95 grams, 1414.61 ± 136.81 grams, 1600.37 ± 147.85 grams;
The average weight in 6,8,10,12,14,16 week age of slow elongated GG strain cock is: 488.78 ± 44.50 grams, 657.87 ± 60.34 grams, 874.45 ± 69.91 grams, 1044.07 ± 101.19 grams, 1203.39 ± 125.67 grams, 1429.30 ± 123.71 grams.
Can find out, fast each stage body weight of elongated AA strain cock exceeds than slow elongated GG strain cock respectively: 7.71%, 7.38%, 8.08%, 7.01%, 17.55%, 11.91%, and difference is extremely remarkable.
The average weight in 6,8,10,12,14,16 week age of fast elongated AA strain hen is: 467.28 ± 35.22 grams, 615.01 ± 50.90 grams, 789.01 ± 69.26 grams, 983.67 ± 80.35 grams, 1133.71 ± 101.50 grams, 1205.18 ± 110.66 grams;
The average weight in 6,8,10,12,14,16 week age of slow elongated GG strain hen is: 409.12 ± 32.27 grams, 543.29 ± 47.26 grams, 704.42 ± 53.77 grams, 856.95 ± 67.64 grams, 978.07 ± 73.91 grams, 1056.03 ± 81.43 grams.
Can find out, fast each stage body weight of elongated AA strain hen exceeds than slow elongated GG strain hen respectively: 14.22%, 13.20%, 12.01%, 14.79%, 15.91%, 14.12%, and difference is extremely remarkable.
Claims (1)
1. a method of setting up the fast elongated AA of limit chicken strain and slow elongated GG strain colony, is characterized in that: comprise the steps:
In the 7th generation numb plumage leaf-comb strain limit chicken group of closed population family breeding, use the PCR-RFLP technology to be detected the genotype in MSTN gene G2283A site, the PCR product is cut with the BbvI enzyme, and the sample called after that will contain 162bp purpose fragment is grown frequency of genotypes AA soon; The sample called after that will contain 127bp purpose fragment is grown genotype GG slowly;
Then, choose respectively fast long frequency of genotypes AA and the male and female chicken group breeding of long genotype GG slowly, turn out respectively the fast elongated AA of limit chicken strain colony and the slow elongated GG of limit chicken strain colony.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104450916A (en) * | 2014-12-11 | 2015-03-25 | 武汉市畜牧兽医科学研究所 | Application of haplotype molecular marker in duck carcass traits detection |
| CN106367519A (en) * | 2016-09-30 | 2017-02-01 | 扬州大学 | Method for improving slaughter performance of Jinghai yellow chicken by MSTN genes |
| CN106386683A (en) * | 2016-09-21 | 2017-02-15 | 佛山科学技术学院 | A cultivation method for a disease-resistant line of Qingyuan partridge chickens without endogenous avian leukosis ev 21 viruses |
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| CN102251051A (en) * | 2011-08-15 | 2011-11-23 | 扬州大学 | Kit for detecting Bian chicken MSTN genotype |
| CN102321750A (en) * | 2011-08-15 | 2012-01-18 | 扬州大学 | Method for rapidly screening bian chicken weight gain degree by molecular marking |
| CN102352410A (en) * | 2011-10-07 | 2012-02-15 | 扬州大学 | Method for rapidly improving number of eggs produced by Bian chicken through DNA (deoxyribonucleic acid) labeling |
| CN103122384A (en) * | 2013-01-23 | 2013-05-29 | 河南农业大学 | Breeding method of laying hen with low egg yolk cholesterol content |
| CN103120143A (en) * | 2013-01-23 | 2013-05-29 | 河南农业大学 | Cultivating method of laying hen with low egg yolk cholesterol content |
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102251051A (en) * | 2011-08-15 | 2011-11-23 | 扬州大学 | Kit for detecting Bian chicken MSTN genotype |
| CN102321750A (en) * | 2011-08-15 | 2012-01-18 | 扬州大学 | Method for rapidly screening bian chicken weight gain degree by molecular marking |
| CN102352410A (en) * | 2011-10-07 | 2012-02-15 | 扬州大学 | Method for rapidly improving number of eggs produced by Bian chicken through DNA (deoxyribonucleic acid) labeling |
| CN103122384A (en) * | 2013-01-23 | 2013-05-29 | 河南农业大学 | Breeding method of laying hen with low egg yolk cholesterol content |
| CN103120143A (en) * | 2013-01-23 | 2013-05-29 | 河南农业大学 | Cultivating method of laying hen with low egg yolk cholesterol content |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104450916A (en) * | 2014-12-11 | 2015-03-25 | 武汉市畜牧兽医科学研究所 | Application of haplotype molecular marker in duck carcass traits detection |
| CN106386683A (en) * | 2016-09-21 | 2017-02-15 | 佛山科学技术学院 | A cultivation method for a disease-resistant line of Qingyuan partridge chickens without endogenous avian leukosis ev 21 viruses |
| CN106367519A (en) * | 2016-09-30 | 2017-02-01 | 扬州大学 | Method for improving slaughter performance of Jinghai yellow chicken by MSTN genes |
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Application publication date: 20131204 |