CN108546765A - A kind of and the relevant SNP marker of Gaoyou duck muscle growth development character, its acquisition methods and application - Google Patents
A kind of and the relevant SNP marker of Gaoyou duck muscle growth development character, its acquisition methods and application Download PDFInfo
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- CN108546765A CN108546765A CN201810449436.7A CN201810449436A CN108546765A CN 108546765 A CN108546765 A CN 108546765A CN 201810449436 A CN201810449436 A CN 201810449436A CN 108546765 A CN108546765 A CN 108546765A
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Abstract
The invention discloses a kind of and the relevant SNP marker of Gaoyou duck muscle growth development character, its acquisition methods and application, the nucleotide sequence such as SEQ ID NO of the molecular labeling:Shown in 1, TNNI1 genes are come from, the 130th of the sequence is SNP mutation site, and the base in the site is A or G.Weight, the chest leg flesh weight correlated traits for the Gaoyou duck that SNP marker through the invention is selected are improved, and can be used for the Seedling selection in family selective breeding, molecule foundation can be also provided in apolegamy.The SNP marker of the present invention be screen Gaoyou duck weight, chest leg flesh provides new method again.
Description
Technical field
The invention belongs to the technical fields of poultry genetic engineering, are sent out with Gaoyou duck muscle growth more particularly, to a kind of
The relevant SNP marker of fertility shape, its acquisition methods and application.
Background technology
China is the first big country of meat duck production and consumption in the world.Duck fat content is low, unsaturated fatty acid content
More, rigidity growth is presented in the demand of duck product in China's meat product total demand.But annual market still has 7-8 hundred million to lack
Mouthful, it differs greatly with development speed, the utilization of resources and the development scale of China's duck industry.Currently, by the state of Beijing duck success selection and breeding
(70 age in days average weights are about 3.3kg, and chest muscle rate is 17%, and leg flesh rate is 9%) meat at home for outer introduced variety cherry valley duck
Duck market has overwhelming superiority, and the special duck in south French in recent years is also entered China market, and the excellent Germplasm Resources of Local in China is not
Rational digging utilization can be obtained, the waste of domestic excellent Germplasm Resources of Local is caused.In meat duck breeding, selectivity improves life
The growth and development of long early stage meat duck skeletal muscle is the barrier of breeding success and industrial upgrading.With modern biotechnology and information
The development of technology, molecular breeding are more and more applied to improvement livestock and poultry species, accelerate Advances in Breeding.
TNNI1 genes belong to the subunit of Troponin I (Troponin I, TnI).TnI is positioned at striated muscle, main work(
Can be by with Ca2+In conjunction with adjusting the activity of myosin ATPase, and then adjust the contraction of muscle.TnI genes include three kinds
Hypotype is divided into skeletal muscle type troponin (TNNI1), quick skeletal muscle type flesh at a slow speed according to Tissue distribution and expression specificity
Calcium protein I (TNNI2) and myocardium type Troponin I (TNNI3).TNNI1 genes are mainly in the slow shrinkage bone of the manhood of people
It is specific expressed in bone flesh.It is on pig studies have shown that TNNI1 has expression in Pig embryos phase different type muscle fibre, fast white
In muscle based on muscle fibre, peak expression appears in 120 days, and in the muscle based on slow switch fibers, peak expression appears in
60 days；It is on sheep studies have shown that expression quantity of the TNNI1 genes in muscle with the increase at age in the trend that is gradually increasing,
Expression quantity is minimum at half year old, tends to balance after 2 years old.It is on sheep studies have shown that TNNI1 gene expression amounts and sheep muscle fibre
Diameter, cross-sectional area etc. have correlation.These reports have all convincingly demonstrated muscle development and life of the TNNI1 genes in animal
Key player is play in long development.Seminar's early-stage study shows that TNNI1 mrna expression amounts and duck chest muscle and leg flesh flesh are fine
The diameter and cross-sectional area of dimension are all without significantly correlated.TNNI1 genes birds there may be different from mammal function with
The mode of action.
Embryonic period, embryonic phase is the important period that animal muscle is formed, and the quantity of myofibroblasts is substantially stationary to get off, number after birth
Amount is not further added by.The increase of meat yield is mainly the increase of diameter of muscle fiber and length after birth.Chest muscle and leg flesh are that duck is main
Production meat position.Currently, there is no the relevant report of TNNI1 gene genetics variation and duck muscle growth development character.Gaoyou duck is
One of national three big ducks, growth and development is fast, with good meat quality, belongs to meat egg dual-purpose type local varieties.In breeding work, with change
Individual inheritance characteristic (induced mutations) compares, and job change genetic structure is still relatively easy to.And in existing gene
On the basis of, improve the genetic structure of kind by changing gene frequency, potentiality or prodigious.
Single nucleotide polymorphism (single nucleotide polymorphism, SNP), is primarily referred to as in genome water
Flat DNA sequence polymorphism caused by a single nucleotide variation.SNP as third generation molecular labeling, have quantity it is more,
The features such as distribution is wide, representativeness is strong and genetic stability is good.Make at present in the analysis of genetic diversity of animals and plants, gene
It is widely used in the research of figure, molecular mark and functional gene.
Therefore, research and the relevant SNP marker of Gaoyou duck muscle growth development character, are used for the choosing of Gaoyou duck early stage
It educates, has great importance.
Invention content
The purpose of the present invention is to provide a kind of with the relevant SNP marker of Gaoyou duck muscle growth development character, it
Acquisition methods and application are used as molecular labeling by finding with the SNP sites of Gaoyou duck muscle growth trait associations, and by its with
Weight and muscle increase again is associated analysis, accelerates breeding process.
Technical scheme is as follows：
A kind of and relevant SNP marker of Gaoyou duck muscle growth development character, the nucleotides sequence of the molecular labeling
Row such as SEQ ID NO:Shown in 1, TNNI1 genes are come from, the 130th of the sequence is SNP mutation site, the base in the site
For A or G.
The present invention also provides described in a kind of acquisition with the relevant SNP marker of Gaoyou duck muscle growth development character
Method, this approach includes the following steps：
(1) PCR amplification Gaoyou duck genomic DNA obtains pcr amplification product；
(2) pcr amplification product is sequenced, genotype is judged by sequence analysis；
(3) different genotype and Gaoyou duck muscle growth development character are associated analysis, to obtain the SNP points
Son label.
It needs to further illustrate, in step (1), such as SEQ ID NO of forward primer sequence used in PCR amplification:Shown in 2,
Reverse primer sequences such as SEQ ID NO:Shown in 3.
In step (2), the genotype is AA, AG or GG.
In step (3), Gaoyou duck muscle growth development character includes weight, chest muscle weight and leg flesh weight.
It is high in screening with the relevant SNP marker of Gaoyou duck muscle growth development character that invention further provides described
Application in postal duck weight, chest muscle weight and leg flesh principal characteristic shape.
Beneficial effects of the present invention：Divide with the relevant SNP of Gaoyou duck muscle growth development character the present invention provides a kind of
Son label, for the SNP polymorphisms of TNNI1 genes, designs the genetic fragment that specific primer amplification includes SNP site, then
Sequencing analysis is carried out, it being capable of simple, quick, low cost, the polymorphism for accurately detecting its mononucleotide.Through the invention
Weight, the chest leg flesh weight correlated traits for the Gaoyou duck that SNP marker is selected are improved, and can be used for family choosing
Seedling selection in educating can also provide molecule foundation in apolegamy.The SNP marker of the present invention be screening Gaoyou duck weight,
Chest leg flesh provides new method again.
Description of the drawings
Fig. 1 is the genotypic results figure in the embodiment of the present invention.
Specific implementation mode
The present invention is carried out with reference to embodiment explanation is explained in detail.
The present invention's is located at the relevant SNP marker of Gaoyou duck muscle growth development character from TNNI1 genes
The 1397382nd of TNNI1 genes, nucleotide sequence such as SEQ ID NO:Shown in 1, the 130th of the sequence is SNP mutation
The base in site, the site is A or G.
For above-mentioned SNP marker, the invention also discloses its acquisition methods, include the following steps：
(1) PCR amplification Gaoyou duck genomic DNA obtains pcr amplification product；
(2) pcr amplification product is sequenced, genotype is judged by sequence analysis；
(3) different genotype and Gaoyou duck muscle growth development character are associated analysis, to obtain the SNP points
Son label.
Wherein, in step (1), such as SEQ ID NO of forward primer sequence used in PCR amplification:Shown in 2, reverse primer sequences
Such as SEQ ID NO:Shown in 3.In step (2), the genotype is AA, AG or GG.In step (3), Gaoyou duck muscle growth hair
Fertility shape includes weight, chest muscle weight and leg flesh weight.
The method of the acquisition and the relevant SNP marker of Gaoyou duck muscle growth development character of the present invention, specifically includes
Following steps：
1, the extraction of the acquisition of duck muscle samples and DNA：
The duck of the present invention comes from Gaoyou duck group of Jiangsu Province, the same age in days Gaoyou duck kind fed under identical daily ration level
Group.After Gaoyou duck is weighed, about 1mL blood is extracted from vein under duck wing using disposable syringe, is injected through high pressure sterilization simultaneously
1.5mL equipped with 2% sterile EDTA (Ethylene diamine tetraacetic acid, EDTA) anti-coagulants of about 200 μ l
It in centrifuge tube, gently shakes up, records wing number, -80 DEG C save backup.Simultaneously duck is butchered after, remove respectively unilateral pectoralis major and
Caput laterale musculi gastrocnemii is simultaneously weighed.Duck is carried out with reference to the specification of blood/cell/tissue DNA extraction kit (Tiangeng, DP304)
The acquisition of blood DNA sample.
2, segment where PCR amplification SNP site
According to TNNI1 genome sequences, design includes the primer of site sequence to be measured, the forward primer and reverse primer
Nucleotide sequence such as SEQNO:Using above-mentioned Gaoyou duck genome as template shown in 2-3, including the positive and negative of site sequence to be measured
It in the presence of primer, Taq archaeal dna polymerases, buffer environment, dNTPs, is expanded under PCR reaction conditions, PCR productions
Object size is 389bp.The primer pair sequence is as follows：
Forward primer sequence is：5’-CTCAGGGTCAGCAGTGTCC-3’(SEQ ID NO：2)；
Reverse primer sequences are：5’-CTCAGCAGCTACACCGAGG-3’(SEQ ID NO：3)
Pcr amplification reaction the specific steps are：
1) pcr amplification reaction system prepares：1 μ l of 50ng/ μ l DNA profilings, 2 μ l of 2mM dNTP, 3mM Mg2+0.6μl、10
2 μ l of × PCR reaction buffers, each 0.4 μ l of 10 μM of forward and reverse primers, 0.2 μ l of 1U/ μ l Taq polymerases, add ddH2O is to total volume
20μl。
2) pcr amplification reaction program is arranged：Expanded on Perkin-Elmer GeneAmp PCR Systems 9600
Increase reaction, pcr amplification reaction program is：95 DEG C of denaturation 2min, 40 cycles (94 DEG C of 90s, 50 DEG C of 1min 30s, 65 DEG C of 30s)
65 DEG C of extension 10min, obtain pcr amplification product.
3) 2 μ l reaction products electrophoresis in 3.0% agarose gel, 0.5 × TBE after reaction, is taken, whether is detection reaction
Success, remaining reaction product preserve under conditions of -20 DEG C.
3, genotype judgement and association analysis
Pcr amplification product is served marine growth Engineering Co., Ltd to be sequenced.According to the result judgement of the sequencing site
Genotype in detecting group.Genotypic results as shown in Figure 1, if the sample is homozygote, just only there are one
Product peak：The peaks A or the peaks G.If it is heterozygosis, then just will appear 2 peaks：The peaks A and the peaks G.
Association analysis has been carried out again to TNNI1 gene polymorphics site different genotype and duck weight, chest muscle weight and leg flesh, has been closed
It is as shown in table 1 to join analysis result, SEQ ID NO：The 130th on 1 is heavy, leg flesh with the weight of 70 age in days of Gaoyou duck, chest muscle
Caput laterale musculi gastrocnemii is in significantly correlated (P again<0.05)；Wherein frequency of genotypes AA is in the weight of 70 ages in days, chest muscle weight, leg flesh gastrocnemius
Lateral head is significantly higher than genotype AG and GG (P again<0.05), so AA genotype is preponderant genotype.In breeding screening, only
AA types are selected, the genotype can be fixed in group.
1 different genotype of table and duck weight, the association analysis of chest muscle weight and leg flesh weight
Note：Lowercase letter indication difference significantly (P in the same row<0.05), same letter indicates difference not significantly (P>
0.05)。
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Any modification, equivalent replacement and simple modifications etc., should all be included in the protection scope of the present invention made by content.
Sequence table
<110>Jiangsu Inst. of Fowls Science
<120>A kind of and the relevant SNP marker of Gaoyou duck muscle growth development character, its acquisition methods and application
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 389
<212> DNA
<213>Duck (Anas platyrhynchos)
<400> 1
ctcagggtca gcagtgtcct gctacacaca ggggcatctt ctgcaaggca ctgactcccc 60
ccctgccctc ctcccgcaga ttaaggacct gaaaatcaaa gtgcttgacc tgcgggggaa 120
gtttaagcga ccgcccctgc ggcgggtccg cgtctccgcc gatgccatgc tgagggctct 180
gctgggctcc aagcacaagg tctccatgga cctcagagcc aacctgaaat ccgtcaagaa 240
ggaggacacg gagaaggtgg gtcgctcccc cagaggcaga gagggctccc gggattgtcc 300
tcgcccttct ccaaggtccc ctgggaagcc tcgagggttt tccaggtacc cagcaggggt 360
actgcagccc cctcggtgta gctgctgag 389
<210> 2
<211> 19
<212> DNA
<213>Artificial sequence (unknown)
<400> 2
ctcagggtca gcagtgtcc 19
<210> 3
<211> 19
<212> DNA
<213>Artificial sequence (unknown)
<400> 3
ctcagcagct acaccgagg 19
Claims (6)
1. a kind of and relevant SNP marker of Gaoyou duck muscle growth development character, which is characterized in that the molecular labeling
Nucleotide sequence such as SEQ ID NO:Shown in 1, TNNI1 genes are come from, the 130th of the sequence is SNP mutation site, the position
The base of point is A or G.
2. a kind of obtaining the method described in claim 1 with the relevant SNP marker of Gaoyou duck muscle growth development character,
It is characterized in that, this approach includes the following steps：
(1) PCR amplification Gaoyou duck genomic DNA obtains pcr amplification product；
(2) pcr amplification product is sequenced, genotype is judged by sequence analysis；
(3) different genotype and Gaoyou duck muscle growth development character are associated analysis, to obtain the SNP molecules mark
Note.
3. the according to claim 2 and relevant SNP marker of Gaoyou duck muscle growth development character acquisition methods,
It is characterized in that, in step (1), such as SEQ ID NO of forward primer sequence used in PCR amplification:Shown in 2, reverse primer sequences are such as
SEQ ID NO:Shown in 3.
4. the according to claim 2 and relevant SNP marker of Gaoyou duck muscle growth development character acquisition methods,
It is characterized in that, in step (2), the genotype is AA, AG or GG.
5. the according to claim 2 and relevant SNP marker of Gaoyou duck muscle growth development character acquisition methods,
It is characterized in that, in step (3), Gaoyou duck muscle growth development character includes weight, chest muscle weight and leg flesh weight.
6. described in claim 1 screening Gaoyou duck body with the relevant SNP marker of Gaoyou duck muscle growth development character
Application in weight, chest muscle weight and leg flesh principal characteristic shape.
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| CN201810449436.7A CN108546765B (en) | 2018-05-11 | 2018-05-11 | SNP molecular marker related to Gaoyou duck muscle growth and development traits as well as acquisition method and application thereof |
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| CN108546765A true CN108546765A (en) | 2018-09-18 |
| CN108546765B CN108546765B (en) | 2021-07-23 |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112852979A (en) * | 2021-03-25 | 2021-05-28 | 中国农业科学院北京畜牧兽医研究所 | Molecular marker related to duck breast muscle fiber diameter character and application thereof |
| CN113373240A (en) * | 2021-06-15 | 2021-09-10 | 扬州大学 | Method for identifying gene locus related to duck muscle development |
| CN113388686A (en) * | 2021-07-21 | 2021-09-14 | 江苏省家禽科学研究所 | SNP locus combination for identifying Gaoyou duck varieties and application thereof |
| CN113481305A (en) * | 2021-08-10 | 2021-10-08 | 安徽农业大学 | Molecular marker for identifying duck leg muscle character based on MyoD gene and method and application thereof |
| CN116064841A (en) * | 2022-11-15 | 2023-05-05 | 昆山市畜牧兽医站 | SNP locus related to growth and development of Loranthus duck and application thereof |
-
2018
- 2018-05-11 CN CN201810449436.7A patent/CN108546765B/en active Active
Non-Patent Citations (2)
| Title |
|---|
| JINGTING SHU: "Molecular Cloning, Characterization, and Temporal Expression Profile of Troponin I Type 1 (TNNI1) Gene in Skeletal Muscle During Early Development of Gaoyou Duck (Anas Platyrhynchos Domestica)", 《ANIMAL BIOTECHNOLOGY》 * |
| Z. Y. XU: "Identification of three novel SNPs and association with carcass traits in porcine TNNI1 and TNNI2", 《MOL BIOL REP 》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112852979A (en) * | 2021-03-25 | 2021-05-28 | 中国农业科学院北京畜牧兽医研究所 | Molecular marker related to duck breast muscle fiber diameter character and application thereof |
| CN113373240A (en) * | 2021-06-15 | 2021-09-10 | 扬州大学 | Method for identifying gene locus related to duck muscle development |
| CN113373240B (en) * | 2021-06-15 | 2024-01-23 | 扬州大学 | Method for identifying gene locus related to duck muscle development |
| CN113388686A (en) * | 2021-07-21 | 2021-09-14 | 江苏省家禽科学研究所 | SNP locus combination for identifying Gaoyou duck varieties and application thereof |
| CN113481305A (en) * | 2021-08-10 | 2021-10-08 | 安徽农业大学 | Molecular marker for identifying duck leg muscle character based on MyoD gene and method and application thereof |
| CN116064841A (en) * | 2022-11-15 | 2023-05-05 | 昆山市畜牧兽医站 | SNP locus related to growth and development of Loranthus duck and application thereof |
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