CN110468212A - SNP site relevant to fatty acid C14:0 content and application on No. 19 chromosomes of meat Simmental - Google Patents
SNP site relevant to fatty acid C14:0 content and application on No. 19 chromosomes of meat Simmental Download PDFInfo
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- CN110468212A CN110468212A CN201910564709.7A CN201910564709A CN110468212A CN 110468212 A CN110468212 A CN 110468212A CN 201910564709 A CN201910564709 A CN 201910564709A CN 110468212 A CN110468212 A CN 110468212A
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Abstract
The present invention provides SNP site relevant to fatty acid C14:0 content and applications on No. 19 chromosomes of meat Simmental, the site of the SNP marker is international ox with reference to the 51333374th nucleotide site on No. 19 chromosomes of genome UMD3.1 version, and the base in the site is T or A.The present invention passes through the advantage allele of the preferably SNP, can be increased generation after generation advantage gene frequency, improves the fatty acid C14:0 content of meat Simmental, accelerates the remains of cattle and passes Improvement advance to effectively improve the economic benefit of Genetic Improvement of Beef Cattle.
Description
Technical field
The present invention relates to SNP site relevant to fatty acid character and its use on No. 19 chromosomes of meat Simmental
On the way.
Background technique
China has nearly 20 years time to the cultivation of meat Simmental.Currently, meat Simmental accounts for China
50% or more of the 60%-70% of all kinds of crossbreeding and improvement cows bodies, Zhan Zhongguo beef cattle livestock on hand ratio.And China's beef cattle progeny testing
System is just begun setting up, it is still necessary to which therefore gradual perfection to accelerate China's Molecular Breeding in Beef Cattle process, is closed using full-length genome
Connection analyzes and identifies the significant site for influencing the fatty acid character of the meat Simmental in China.The content of fatty acid of muscle is assessment
The important indicator of meat, ingredient are of great significance to the promotion of meat flavor, and meat flavor be often consumer most
For one of the principal element for the evaluation meat valued.Have result of study show fatty acid composition tend to belong in low genetic force
Character, and the genetic force of C14:0 content is relatively high, about 0.54.Therefore, using genetic approach to the meat west of sources group
It is feasible that door Simmental Cattle Chromosome fatty acid C14:0 content, which carries out improvement,.Fatty acid C14:0 content is the quantity controlled by multiple genes
There are a large amount of quantitative trait locus (quantitative trait for influencing fatty acid C14:0 contents in cow genome group in character
Loci, QTLs).Currently, utilizing the side of whole-genome association (genome-wide association study, GWAS)
Method differentiates to be more than many QTLss relevant to fatty acid C14:0 content and a large amount of mononucleotide polymorphic positions in cow genome group
SNP wherein with active effects is added to molecule mark by point (single nucleotide polymorphisms, SNPs)
In note assisted Selection (marker assisted selection, MAS), gene group selection (Genomic selection, GS)
The genetic improvement progress of fatty acid C14:0 content can be then significantly improved, and then improves offspring beef cattle fatty acid C14:0 content, is mentioned
High beef meat flavor enhances the market competitiveness of breeding enterprise commodity Beef production.
Whole-genome association is to identify influence table by statistical analysis strategy based on the linkage disequilibrium between SNP
The analysis method of relationship between type and genotype has played important in terms of identifying the molecular labeling for influencing ox important economical trait
Effect.The present invention has differentiated the significant of the meat Simmental fatty acid C14:0 content of influence by GWAS analysis strategy
SNP is used in molecular marker assisted selection and gene group selection, and selection is to the raising advantageous base of fatty acid C14:0 content
Because type is reserved seed for planting, to improve the gene frequency of advantage allele by generation, then it can accelerate kind of a process for ox breeding improvement, be
Beef cattle breeding brings great economic benefit.
Summary of the invention
In order to achieve the above objectives, the primary purpose of the present invention is that provide No. 19 chromosomes of ox on meat Simmental
The relevant SNP site of beef fat acid C14:0 content, the nucleotide sequence of the molecular labeling as shown in SEQ ID NO:1,
M in middle sequence is T or A, leads to the difference of beef fat acid C14:0 content.
The molecular labeling is located on the nucleotide sequence on No. 19 chromosomes of meat Simmental, the molecule mark
The SNP site of note is the coding mutation for the T119-A119 that SEQ IDNO:1 sequence labelling position is 119；The molecule
The SNP site of label corresponds to the 51333374th T > A on international cow genome group UMD3.1 version No. 19 chromosomes of reference sequences
Mutation.
It is individual in the ox of screening high fatty acid C14:0 content that another object of the present invention is to provide above-mentioned molecular labelings
Method, specifically, molecular labeling described in claim 1 on detection No. 19 chromosomes of ox, 5 ' ends the of the molecular labeling
119 mononucleotides are T or A, eliminate A and retain T.It is embodied in the ox and is selected from In Xilingol League In Inner Mongolia crow drawing lid pipe
The meat Simmental sources group in the pasture Li Qu.
Another object of the present invention is to provide a kind of for identifying the molecule mark of above-mentioned influence beef fat acid C14:0 content
The nucleic acid sequence of the primer pair of note, the primer pair is as follows:
Forward primer sequence is as shown in SEQ ID NO:2；
Reverse primer sequences are as shown in SEQ ID NO:3.
Above-mentioned primer pair influences the application in beef fat acid C14:0 content in identification.
Application of the above-mentioned primer pair in cow genome group selection.
Above-mentioned primer pair is improving the application in beef fat acid C14:0 content.
The purpose of the present invention is to provide a kind of methods of the genetic improvement of ox, which comprises determines kind of ox resource
The site of the molecular labeling of the above-mentioned influence beef fat acid C14:0 content of kind ox in group, and according to that molecular marker
It makes corresponding selection: selecting international ox with reference on No. 19 chromosomes of genome UMD3.1 version in described kind of ox sources group
51333374th site is TT and the kind ox individual of TA, AA genotype, and eliminating in the 51333374th site is AA genotype
Ox individual, to improve the frequency of the allele T in the site by generation, to improve the fatty acid C14:0 content of offspring ox.
The present invention has the following advantages and effects with respect to the prior art:
Present invention research simultaneously determines molecular labeling relevant to beef fat acid C14:0 content, verifies it to fatty acid C14:0
The influential effect of content finally establishes the genome selection and use technology of efficiently and accurately, is applied to kind of ox and improves fatty acid
In the genetic improvement of C14:0 content, to improve the fatty acid C14:0 of offspring ox, meat is improved, and then increase breeding enterprise
The market competitiveness.
Detailed description of the invention
Fig. 1 is that full-length genome association of the meat Simmental on No. 19 chromosomes about fatty acid C14:0 content divides
Analyse the Manhattan (GWAS) figure；Wherein: the chromosome numbers of abscissa expression ox；Ordinate expression-logP value.
Fig. 2 is the fatty acid C14:0 content of the meat Simmental of different genotype.
Specific embodiment
Present invention will now be described in further detail with reference to the embodiments and the accompanying drawings, but embodiments of the present invention are unlimited
In this.
Foregoing invention purpose of the invention is specifically achieved in that
Embodiment 1
1, experimental animal
Experiment cows group used in the present invention is all from In Xilingol League In Inner Mongolia crow and draws lid directorial area pasture China
Meat Simmental 663 is set up for Institute of Animal Sciences, Chinese Academy of Agricultural Sciences's cattle genetics and breeding innovation team
Meat Simmental sources group.
663 meat Simmentals in the sources group are chosen in this experiment altogether.Meat Simmental sources group is every
Carry out group expanding year, newly-increased individual is general to be undergone birth, fattens and butcher 3 stages.The calf being born the 3-5 month every year is by one
The section time put management in a suitable place to breed after, unified birth weight and body measurement are carried out by cattle genetics and breeding innovation team in July in the same year, and
Basic cow is measured simultaneously.The young ox in 5~September age fatten in Unified Set by October in the same year, and collects life
The phenotypic data of long development character, while left and right obtains genotype data for Illumina BovineHD chip gene parting.
When all individual fattening periods reach 10-12 months, i.e. in or so the November in the coming year, all meat Simmentals are carried out in batches
It butchers.Slaughtering process in strict accordance with " meat purchase specifications " execute, butcher data, trunk data and Meat quality data in strict accordance with
The requirement of GB/T 27643-2011 " butchering rear carcass trait and Meat Quality measurement guide " is measured.
2, sample collection
All individual venous blood 50ml of above-mentioned cows body are collected with heparin tube, -80 DEG C of refrigerators is placed in and saves backup.3, ox
Full-length genome 770K superchip SNP sentences type
Each of 663 meat Simmentals chosen from above-mentioned sources group individual acquisition venous blood 50ml,
Complete genome DNA is extracted with standard phenol-chloroform method, it is every through Nanodrop2000/2000C nucleic acid-protein detector Accurate Determining
The concentration and OD ratio (OD260/280, OD260/230) of the DNA of a sample.Through NanoDrop2000/2000C nucleic acid egg
DNA is diluted to 50ng/ μ L or so according to the concentration of detection by the DNA sample of white detector test qualification.6 μ l have been extracted again
Good DNA sample to be measured is mixed with 2 μ l Loading Buffer, is loaded in 1% Ago-Gel, electric under 150V voltage
Swim 25min, observes and takes pictures under UV detector and gel imaging equipment, observes the integrality of DNA.
DNA sample Song Niuqin biotechnology (Shanghai) Co., Ltd. carries out ox full-length genome according to company standard process
Illumina BovineHD chip 770K SNP chip (Illumina, the U.S.) genotype determines.It is soft using PLINK v1.90
Part carries out quality control to all sample 770K chip scanning typing datas, rejects and detects individual rate lower than 90%, family Meng De
Your error rate is higher than 0.1, minimum gene frequency less than 0.05 and Hardy-Weinberg equilibrium significance is higher than 10-6's
SNP finally obtains the effective gene type data of 671,204 SNP.
4, full-length genome association (GWAS) analysis
In order to eliminate group's stratification effect, the present invention is using the regression analysis of linear mixed model single-point and combines R language
GenABEL software package carries out GWAS analysis, and the similarity correction stratification effect of genome between individual is utilized in analysis model.Using
Bonferrini method determines the conspicuousness threshold value of SNP Yu fatty acid C14:0 content correlation degree, and genomic level remarkable threshold is
0.05 divided by effective SNP site quantity, i.e., genome level of signifiance threshold value is 7.45e-8, i.e., 0.05/671,204 (effective SNP
Quantity)；Chromosome level remarkable threshold is 1 divided by effective SNP site quantity, i.e. chromosome level of signifiance threshold value is 1.49e-6,
I.e. 1/671,204 (effective SNP quantity).
It is as shown in Figure 1 that GWAS analyzes result.From fig. 1, it can be seen that existing in No. 19 chromosomes of meat Simmental significant
The site of fatty acid C14:0 content is influenced, most strongly connected SNP is g.119T > A (P=1.80E-10).
5, the association analysis of different genotype and fatty acid C14:0 content phenotype
According to table 1, the SNP site of molecular labeling g.119T > A and extremely significant related (the P < of fatty acid C14:0 content
0.001), illustrate that this molecular labeling significantly affects the fatty acid C14:0 content of ox, the auxiliary of this SNP site to ox can be passed through
Selection is helped, to improve the fatty acid C14:0 content of the group, and then accelerates the breeding process of objective trait.
According further to table 1 it is found that the fatty acid C14:0 content of TT type, TA type ratio AA type is high, illustrate that AA type ox individual is right
Screening fatty acid C14:0 content height is unfavorable, therefore preferentially retains the kind ox of TT, TA type.Fatty acid C14:0 content is to measure beef matter
Important indicator, improve ox fatty acid C14:0 content be conducive to improve beef meat flavor, and then increase the market competitiveness.
Therefore, it needs to be phased out the kind ox of AA type in breeding process, preferentially retains the kind ox of TT, TA type, to improve the position by generation
The frequency of the allele T of point.
The correlation of the SNP site of 1 molecular labeling of table g.119T > A and fatty acid C14:0 content
6, target DNA sequence amplification and sequencing
(1) design of primers
No. 19 chromosomes of ox are downloaded by the website Ensembl (http://asia.ensembl.org/index.html)
The DNA sequence dna of upper SEQ ID NO:1.And utilize 6.0 design primer of primer-design software primer premier.
The DNA sequence dna of the primer of design is as follows:
P001 is positive: 5 '-CCTGCACAATGAGGGACCAG-3 ',
P002 is reversed: 5 '-TGTCCGACTCTTTGCAACCC-3 '；
(2) PCR amplification
DNA profiling 1uL, distilled water 3.4uL, 2 × Tag PCR StanMix with are added in the reaction system of 10uL
Each 0.3ul of Loading Dye 5uL, primer P001 and P002.PCR reaction condition are as follows: after 94 DEG C of initial denaturation 5min, 94 DEG C of changes
Property 30s, 56 DEG C of annealing 30s, 72 DEG C of extension 45s, 35 circulation, last 72 DEG C of extensions 5min.
(3) determined dna sequence
DNA sequence dna sequencing identification: carrying out in Beijing Sheng Gong Science and Technology Ltd., and genetic fragment surveys positive and negative two reactions.It will
Measured sequence and NCBI genomic sequence comparison, obtains the mutation of corresponding SNP site.Sequencing result is as follows:
Note: the M marked in sequence table is mutational site, (is mutating alkali yl in bracket, is equipotential with display is underlined
Gene mutation), design primer sequence location is shown as in the head and the tail overstriking of the sequence.
7, the SNP site of molecular labeling g.119T > A effect analysis
By being eliminated to the Niu Jinhang that genotype in group is AA, being remarkably improved group to molecular marker assisted selection
Fatty acid C14:0 content bring more economic benefits for enterprise to improve meat.
The present invention tentatively carries out it by detecting to the 119th base mutation site in SEQ ID NO:1 sequence
The application of association analysis between genotype and the fatty acid C14:0 content of ox is the molecular marker assisted selection and gene of ox
Group selection provides a new molecular labeling.
The above embodiment is a preferred embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, other any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present invention,
It should be equivalent substitute mode, be included within the scope of the present invention.
Claims (9)
1. SNP site relevant to fatty acid C14:0 content and application, feature exist on No. 19 chromosomes of meat Simmental
In the site of the SNP marker is international ox with reference to the 51333374th nucleosides on No. 19 chromosomes of genome UMD3.1 version
Sour site, the base in the site are T or A.
2. SNP marker according to claim 1, which is characterized in that the sequence of the SNP marker such as SEQ ID NO:1 institute
Show, the 119th bit base from 5 ' ends of sequence shown in the SEQ ID NO:1 is T or A.
3. a kind of for detecting the primer pair of SNP marker as claimed in claim 1 or 2, which is characterized in that the primer pair
Nucleic acid sequence is as shown in SEQ ID NO:2 and SEQ ID NO:3.
4. a kind of for detecting the kit of SNP marker as claimed in claim 1 or 2, which is characterized in that wanted including such as right
Primer pair described in asking 3.
5. a kind of method for improving meat Simmental fatty acid C14:0 content, which is characterized in that the method includes following
Step:
Detect the gene of the 51333374th nucleotide site on International Reference genome UMD3.1 No. 19 chromosomes of version of ox
Type selects TT, TA type individual of the 51333374th nucleotide site as kind of an ox.
6. according to the method described in claim 5, it is characterized in that, the international ox of the detection ox refers to genome UMD3.1 editions
On this No. 19 chromosome the genotype of the 51333374th nucleotide site method the following steps are included:
(1) genomic DNA of ox to be measured is extracted；
(2) primer pair as claimed in claim 3 is used, the genomic DNA of the ox to be measured is subjected to PCR amplification, to obtain
Pcr amplification product；
(3) pcr amplification product is sequenced, to obtain sequencing result；
(4) it is based on the sequencing result, determines the genotype of the SNP marker as claimed in claim 1 or 2 of the ox to be measured.
7. according to the method described in claim 5, it is characterized in that, the cows body includes meat Simmental and its synthesis
System.
8. SNP marker as claimed in claim 1 or 2 is improving the purposes in beef fat acid C14:0 content.
9. primer pair as claimed in claim 3 or kit as claimed in claim 4 are improving beef fat acid C14:0 content
In purposes.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111705137A (en) * | 2020-03-19 | 2020-09-25 | 中国农业科学院北京畜牧兽医研究所 | SNP (Single nucleotide polymorphism) locus related to live weight of Chinese cattle and cattle before slaughter and application |
| CN112195253A (en) * | 2020-10-28 | 2021-01-08 | 中国农业科学院北京畜牧兽医研究所 | SNP (Single nucleotide polymorphism) locus for increasing content of fatty acid C14:0 in chicken and method for breeding high-quality chicken strain by using SNP locus |
| CN117089636A (en) * | 2023-10-20 | 2023-11-21 | 中国农业大学 | Molecular marker combination for analyzing goat meat performance and application |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111705137A (en) * | 2020-03-19 | 2020-09-25 | 中国农业科学院北京畜牧兽医研究所 | SNP (Single nucleotide polymorphism) locus related to live weight of Chinese cattle and cattle before slaughter and application |
| CN111705137B (en) * | 2020-03-19 | 2021-10-26 | 中国农业科学院北京畜牧兽医研究所 | SNP (Single nucleotide polymorphism) locus related to live weight of Chinese cattle and cattle before slaughter and application |
| CN112195253A (en) * | 2020-10-28 | 2021-01-08 | 中国农业科学院北京畜牧兽医研究所 | SNP (Single nucleotide polymorphism) locus for increasing content of fatty acid C14:0 in chicken and method for breeding high-quality chicken strain by using SNP locus |
| CN117089636A (en) * | 2023-10-20 | 2023-11-21 | 中国农业大学 | Molecular marker combination for analyzing goat meat performance and application |
| CN117089636B (en) * | 2023-10-20 | 2023-12-22 | 中国农业大学 | Molecular marker combination for analyzing goat meat performance and application |
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Effective date of registration: 20211224 Address after: 026321 in hesguula pasture, Ulagai management area, Xilin Gol League, Inner Mongolia Autonomous Region Patentee after: Inner Mongolia okos animal husbandry Co.,Ltd. Address before: 100193 No. 2 Old Summer Palace West Road, Beijing, Haidian District Patentee before: INSTITUTE OF ANIMAL SCIENCES, CHINESE ACADEMY OF AGRICULTURAL SCIENCES |