CN101864486B - Primer for detecting egg laying performance of goose and method and application thereof - Google Patents
Primer for detecting egg laying performance of goose and method and application thereof Download PDFInfo
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- CN101864486B CN101864486B CN201010196932XA CN201010196932A CN101864486B CN 101864486 B CN101864486 B CN 101864486B CN 201010196932X A CN201010196932X A CN 201010196932XA CN 201010196932 A CN201010196932 A CN 201010196932A CN 101864486 B CN101864486 B CN 101864486B
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- goose
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- prl268tt
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- 241000272814 Anser sp. Species 0.000 title claims abstract description 87
- 238000000034 method Methods 0.000 title claims abstract description 21
- 230000036750 egg laying performance Effects 0.000 title claims abstract description 16
- 108090000790 Enzymes Proteins 0.000 claims abstract description 12
- 102000004190 Enzymes Human genes 0.000 claims abstract description 12
- 238000012408 PCR amplification Methods 0.000 claims abstract description 11
- 108091008146 restriction endonucleases Proteins 0.000 claims abstract description 11
- 239000002773 nucleotide Substances 0.000 claims abstract description 4
- 125000003729 nucleotide group Chemical group 0.000 claims abstract description 4
- 238000001514 detection method Methods 0.000 claims description 9
- 230000029087 digestion Effects 0.000 claims description 8
- 238000006062 fragmentation reaction Methods 0.000 claims description 6
- 238000000246 agarose gel electrophoresis Methods 0.000 claims description 3
- 238000013467 fragmentation Methods 0.000 claims description 2
- 238000009400 out breeding Methods 0.000 claims description 2
- 108090000623 proteins and genes Proteins 0.000 abstract description 9
- 238000004519 manufacturing process Methods 0.000 abstract description 4
- 235000013601 eggs Nutrition 0.000 description 11
- 238000009395 breeding Methods 0.000 description 5
- 102000003946 Prolactin Human genes 0.000 description 4
- 108010057464 Prolactin Proteins 0.000 description 4
- 230000001488 breeding effect Effects 0.000 description 4
- 229940097325 prolactin Drugs 0.000 description 4
- 229920002684 Sepharose Polymers 0.000 description 3
- 230000009182 swimming Effects 0.000 description 3
- 239000005557 antagonist Substances 0.000 description 2
- 238000013016 damping Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 238000003384 imaging method Methods 0.000 description 2
- 244000144972 livestock Species 0.000 description 2
- 244000144977 poultry Species 0.000 description 2
- 229910021642 ultra pure water Inorganic materials 0.000 description 2
- 239000012498 ultrapure water Substances 0.000 description 2
- 241000272808 Anser Species 0.000 description 1
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 1
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 1
- 108010042407 Endonucleases Proteins 0.000 description 1
- 102000004533 Endonucleases Human genes 0.000 description 1
- 241000287828 Gallus gallus Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 102000035824 beta Subunit Follicle Stimulating Hormone Human genes 0.000 description 1
- 108010081485 beta Subunit Follicle Stimulating Hormone Proteins 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000004087 circulation Effects 0.000 description 1
- 210000003785 decidua Anatomy 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 210000004907 gland Anatomy 0.000 description 1
- 230000029082 maternal behavior Effects 0.000 description 1
- 210000002394 ovarian follicle Anatomy 0.000 description 1
- 238000005498 polishing Methods 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 210000004291 uterus Anatomy 0.000 description 1
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- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a primer for detecting egg laying performance of goose and method and application thereof. The nucleotide sequence of the primer for detecting egg laying performance of goose is shown by sequence 1 and sequence 2 in the sequence list. The method for detecting egg production of goose includes that gene group DNA of goose to be detected is taken as a template, the DNA molecules shown as sequence 1 and sequence 2 in the sequence list are taken as primers, PCR amplification is carried out, BstYI restriction enzyme restricts the PCR amplification product, and the size of restriction product of BstYI restriction enzyme is detected; the egg production of the PRL268TT gene type goose is higher than that of the PRL268GG gene type goose or the PRL268TG gene type goose. The method for detecting egg laying performance of goose in the invention can determine the gene type of goose to be detected by simple PCR amplification and enzyme restriction, the PRL268TT gene type goose is bred, and candidate goose breed with high egg production can be obtained.
Description
Technical field
The present invention relates to a kind of primer and method and application that detects egg laying performance of goose.
Background technology
China's goose variety source is abundant, its egg productivity from 20 pieces to not waiting more than 100 pieces, cause the production zone of goose to divide obviously, some low yield geese are because the problem of reproductivity and directly influence its kind with being worth and commercial promise.Reproductivity heritability is low, has proved to rely on its progress of conventional selection very slowly.(Prolactin is a kind of proteohormonees of excretory such as prepituitary gland oxyphie, immunocyte and gravid uterus decidua PRL) to prolactin antagonist, belongs to tethelin/prolactin antagonist family.The effect of bird PRL mainly is to suppress growing of ovarian follicle, regulate female bird with regard to nest with maternal behavior such as hatch.Existing research shows, what PRL caused goose influences its egg productivity with regard to nest, and goose is significantly higher than in the PRL level with regard to the nest phase and stops term and laying period, has dependency between plasma prolactin level and the female bird egg productivity.Through molecular breeding method possibly be the effective way that solves an above-mentioned difficult problem.Build that utilizes FSH β assisted Selection goose litter size, dwarf gene assisted Selection chicken at present etc. has made substantial progress; Obtain the functional gene and the mark thereof of livestock and poultry economic characters, and to utilize its breeding of quickening livestock and poultry important economical trait progress be the important innovations in Animal Genetics field.Because the heritability of goose reproductivity is lower, the effect of Phenotypic Selection is very poor always, utilizes molecular biology method to crack the reason stagnation of goose egg productivity variation, does not have substantive breakthroughs.
Summary of the invention
The purpose of this invention is to provide a kind of primer and method and application that can be used to detect egg laying performance of goose.
The primer of detection egg laying performance of goose provided by the present invention, its nucleotide sequence is shown in sequence in the sequence table 1 and sequence 2.
The method of detection egg laying performance of goose provided by the present invention; Be to be template with goose genomic dna to be measured; Is primer with sequence in the sequence table 1 with the dna molecular shown in the sequence 2; Carry out pcr amplification, BstY I digestion with restriction enzyme pcr amplification product, the size of detection BstY I digestion with restriction enzyme product;
Cut product when enzyme and contain a dna fragmentation, size is at 300～400bp, and goose to be measured is the PRL268TT genotype;
Cut product when enzyme and contain three dna fragmentations, size is respectively at 300～400bp, 200～300bp and 100～200bp, and goose to be measured is the PRL268TG genotype;
Cut product when enzyme and contain two dna fragmentations, size is respectively at 200～300bp and 100～200bp, and goose to be measured is the PRL268GG genotype;
The genotypic egg laying performance of goose of said PRL268TT is greater than genotypic goose of said PRL268GG or the genotypic goose of said PRL268TG.
The method of detection egg laying performance of goose of the present invention, wherein: the method for the size of said detection BstY I digestion with restriction enzyme product is an agarose gel electrophoresis.
The method of cultivation goose provided by the present invention is the method with said detection egg laying performance of goose, obtains the genotypic goose of PRL268TT, is that the parent carries out breeding acquisition goose with the genotypic goose of said PRL268TT.
The present invention detects the method for egg laying performance of goose; Can cut the genotype of confirming goose to be measured through simple PCR amplification and enzyme; PRL268TT type egg productivity is significantly higher than PRL268GG type or PRL268TG type (P＜0.01), carries out the high goose strain of egg productivity that breeding can obtain the candidate with the genotypic goose of PRL268TT.
Description of drawings
Fig. 1 is the collection of illustrative plates behind the BstY I digestion with restriction enzyme PCR product.
Embodiment
Embodiment,
One, identifies the PRL268T/G genotype
Extract west, Anhui white goose genomic dna.Genomic dna to extract is a template, and PRLS and PRLA are that primer carries out pcr amplification, and the nucleotide sequence of primer PRLS and PRLA is following:
PRLS：5′-TCTCTTCATCAAACCATACTCAG-3′
PRLA：5′-GTTTCTAGAGTTGTTACCTGTCAGT-3′
Pcr amplification system: total system 25 μ L, 10 μ M/L PRLS, 1.0 μ L, 10 μ M/L PRLA, 1.0 μ L; 10 * PCR damping fluid, 2.5 μ L, 2mM/L dNTPs 2.0 μ L, 2.5U/ μ L Taq archaeal dna polymerase 0.5 μ L; 50ng/ μ L template 1.0 μ L, all the other use the ultrapure water polishing.
94 ℃ of preparatory sex change 5min of pcr amplification condition; 94 ℃ of sex change 30s, 56 ℃ of annealing 30s, 72 ℃ are extended 50s, 35 circulations, last 72 ℃ are extended 10min, 4 ℃ of preservations.
With BstY I restriction enzyme the PCR product being carried out enzyme cuts.
20 μ L endonuclease reaction systems: 100～300ng PCR product, the BstY I restriction enzyme of 5U, 10 * damping fluid of 2.0 μ L adds ultrapure water to 20 μ L.
37 ℃ of digestion 12 hours, 3% agarose gel electrophoresis detects, gel imaging appearance analysis and take pictures the statistics genotype.
The gel imaging result is as shown in Figure 1, and the PCR product after enzyme is cut presents a band on sepharose, and size is 300～400bp (accurately being 384bp), and this goose is PRL268TT genotype (swimming lane 1,3,9,11 and 12 of Fig. 1); The PCR product is consistent with expected results through order-checking.
PCR product after enzyme is cut; On sepharose, present three bands, at 300～400bp (accurately being 384bp), 200～300bp (accurately being 235bp) and 100～200bp (accurately being 149bp), this goose is the PRL268TG genotype (swimming lane 4 of Fig. 1 to the size of three bands respectively; 8,10 and 13); The PCR product is consistent with expected results through order-checking.
PCR product after enzyme is cut presents two bands on sepharose, at 200～300bp (accurately being 235bp) and 100～200bp (accurately being 149bp), this goose is PRL268GG genotype (swimming lane 2,5,6 and 7 of Fig. 1) to the size of two bands respectively; The PCR product is consistent with expected results through order-checking.
Two, the dependency of PRL268T/G genotype and goose egg productivity
The low yield goose: totally 156 of Anhui west white gooses, Lay mattress goose, bright moral goose, Bai Luoman, the high yield goose: totally 182 of Yangzhou white goose, Sichuan white gooses, with the method identified gene type of step (), add up goose egg output simultaneously, the result sees table 1.
In the goose breeding of reality, the ratio of low yield goose PRL268GG type accounts for 83.87%, and the PRL268TT type accounts for 68.13% in the high yield goose.
PRL268TT type egg productivity is significantly higher than PRL268GG type or PRL268TG type (P＜0.01), selects the genotypic goose of PRL268TT to carry out the goose strain that breeding can obtain candidate's high egg productivity.
The lay eggs cognation of level of table 1PRL268T/G different genotype and goose
Above embodiment describes preferred implementation of the present invention; Be not that scope of the present invention is limited; Design under the prerequisite of spirit not breaking away from the present invention; Various distortion and improvement that the common engineering technical personnel in this area make technical scheme of the present invention all should fall in the definite protection domain of claims of the present invention.
Sequence table
< 110>Agricultural University Of Anhui
< 120>primer and the method and the application of detection egg laying performance of goose
<160>2
<210>1
<211>23
<212>DNA
< 213>artificial sequence
<400>1
tctcttcatc?aaaccatact?cag 23
<210>2
<211>25
<212>DNA
< 213>artificial sequence
<400>2
gtttctagag?ttgttacctg?tcagt 25
Claims (4)
1. primer that detects egg laying performance of goose, its nucleotide sequence like sequence in the sequence table 1 with shown in the sequence 2, wherein: said goose is west, Anhui white goose, Lay mattress goose, bright moral goose, Bai Luoman, Yangzhou white goose or Sichuan white goose.
2. method that detects egg laying performance of goose; Being to be template with goose genomic dna to be measured, is primer with sequence in the sequence table 1 with the dna molecular shown in the sequence 2, carries out pcr amplification; BstYI digestion with restriction enzyme pcr amplification product, the size of detection BstYI digestion with restriction enzyme product;
Cut product when enzyme and contain a dna fragmentation, size is at 300～400bp, and goose to be measured is the PRL268TT genotype;
Cut product when enzyme and contain three dna fragmentations, size is respectively at 300～400bp, 200～300bp and 100～200bp, and goose to be measured is the PRL268TG genotype;
Cut product when enzyme and contain two dna fragmentations, size is respectively at 200～300bp and 100～200bp, and goose to be measured is the PRL268GG genotype;
The genotypic goose egg productivity of said PRL268TT is greater than genotypic goose of said PRL268GG or the genotypic goose of said PRL268TG;
Said goose is west, Anhui white goose, Lay mattress goose, bright moral goose, Bai Luoman, Yangzhou white goose or Sichuan white goose.
3. method according to claim 2 is characterized in that: the method for the size of said detection BstYI digestion with restriction enzyme product is an agarose gel electrophoresis.
4. cultivating the method for goose, is to detect goose with claim 2 or 3 described methods, obtains the genotypic goose of PRL268TT, is that the parent carries out breeding acquisition goose with the genotypic goose of said PRL268TT.
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| CN201010196932XA CN101864486B (en) | 2010-06-10 | 2010-06-10 | Primer for detecting egg laying performance of goose and method and application thereof |
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| CN101864486B true CN101864486B (en) | 2012-08-29 |
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| CN107047468B (en) * | 2017-05-05 | 2021-07-13 | 广东海洋大学 | Method for breeding recessive white curly eucheuma chickens |
| CN110283915B (en) * | 2019-07-02 | 2021-08-24 | 华南农业大学 | Method for screening high-egg-laying lion-head geese and application thereof |
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Non-Patent Citations (3)
| Title |
|---|
| 杨涛.鹅GnRH基因5"端调控区和外显子1SNPs检测及与产蛋量的关系.《中国优秀硕士学位论文全文数据库农业科技辑》.2008,(第9期),全文. * |
| 梁忠 等.鹅3_羟基_3_甲基戊二酸单酰辅酶A还原酶_省略_含子5的SNP及其与鹅重要经济的关联性.《农业生物技术学报》.2007,第15卷(第6期),936-941. * |
| 马缨.四川白鹅prl外显子2和5SNP与产蛋性能关系的初探.《中国优秀硕士学位论文全文数据库农业科技辑》.2009,(第10期),全文. * |
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