CN110317880A - Molecular labeling relevant to pannage conversion ratio, identification and its application - Google Patents
Molecular labeling relevant to pannage conversion ratio, identification and its application Download PDFInfo
- Publication number
- CN110317880A CN110317880A CN201910630651.1A CN201910630651A CN110317880A CN 110317880 A CN110317880 A CN 110317880A CN 201910630651 A CN201910630651 A CN 201910630651A CN 110317880 A CN110317880 A CN 110317880A
- Authority
- CN
- China
- Prior art keywords
- pig
- snp
- genotype
- snp marker
- gene
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6809—Methods for determination or identification of nucleic acids involving differential detection
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/124—Animal traits, i.e. production traits, including athletic performance or the like
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/156—Polymorphic or mutational markers
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/172—Haplotypes
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Analytical Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a kind of SNP markers relevant to pannage conversion ratio, what the SNP marker was located at DHRS4 gene includes subregion, No. 7 the 75245571st bit bases of chromosome of specially pig genome version Ensembl Sscrofa 11.1, the SNP marker have A/T polymorphism；The corresponding three kinds of genotype of the SNP marker are respectively TT, TA and AA, and the pig that genotype is AA is TA compared to genotype and the pig of TT has lower feed conversion rate.The determination of DHRS4 gene of the present invention and SNP marker relevant to pannage conversion ratio can be used for identifying or assisting the low pig variety of identification feed conversion rate, shorten breeding cycle, improve breeding efficiency.
Description
Technical field
The present invention relates to pig breeding technical fields, and in particular to molecular labeling relevant to pannage conversion ratio, identification and
It is applied.
Background technique
Animal husbandry in modern agricultural development in occupation of very important status, the ratio that is occupied in agricultural production
It is the important indicator for measuring a countries and regions method development degree.Pork makes as the most important meat sources in China
Pig-breeding industry is obtained in China's animal husbandry in occupation of very important status.In pig breeding industry production, pig economic characters are
Refer to the character for influencing pig productivity, mainly affects the economical production total value of pig breeding industry, and feed cost accounts for pig raising totle drilling cost
65~75%, therefore it is most important to reduce the food consumption in pig-breeding, reduction feed cost.And feed conversion rate is as meat
One of the important indicator of pig economic characters, has become the important directions of research at present.
Feed conversion rate refers to forage volume consumed by every product for increasing 1KG weight, food conversion during performance measurement
Rate is lower, illustrates that the quantity of feed consumed in the product of every production unit weight is fewer, can significantly reduce production cost, increases
The productivity effect of big farm.Feed conversion rate (FCR) is by pork pig feeding process and activity, abilities of digestive and absorption, environment
The influence of the different factors such as temperature adjusting, animal metabolism and body composition, with average daily gain (ADG), the thickness of backfat (BF), residue
Feed intake (RFI) is that the growth performances such as waterpower and carcass lean meat percentage are related with m eatquality, the production effect of strong influence pig
Rate.Gene relevant to feed conversion rate takes part in the numerous metabolic activities of animal body, synthesis, potassium ion generation such as triglyceride
The distribution situation of fat cell, the metabolic activity of liver and animal alimentary canal development etc. are thanked, adjusted, therefore hog feed is converted
The relevant identified for genes of rate can provide important theoretical foundation for the genetic improvement of pork pig growth traits.
Genetic marker assisted selection refers to using the DNA molecular marker with objective trait gene close linkage to mesh
A kind of modern breeding method that character carries out genotype selection is marked, there is operational stability is strong, does not interfere vulnerable to environmental factor etc.
Advantage can greatly improve breeding efficiency.Wherein, how to filter out is with the significantly associated molecular labeling site of objective trait
The key link that assisted selection technology is established.In common molecular marking technique, single nucleotide polymorphism (SNP) application
The most extensively.SNP is third generation genetic marker, is primarily referred to as caused by a single nucleotide variation at the genomic level
DNA sequence polymorphism has many advantages, such as more quantity, widely distributed, high-frequency, low mutation rate, is suitable for quick, scale screening,
It has been applied to genome analysis, biological information automatic detection, simple and complex disease genetic research, herding breeding heredity
Marker research etc..
Therefore, a gene relevant to pannage conversion ratio is excavated, while screening relevant to pannage conversion ratio point
The problem of son label, the identification of the molecular labeling and its application are current urgent need to resolve.
Summary of the invention
In order to overcome the above problem, present inventor has performed sharp studies, first to DHRS4 gene in pig difference growth step
Expression in section skeletal muscle is detected, and is then detected DHRS4 gene polynorphisms and is carried out gene to polymorphic site
Parting, then polymorphic site and the important growth traits of pig are associated analysis, obtained in pig DHRS4 gene for the first time and
The relevant SNP marker of feed conversion rate, the pig variety for identifying or assisting identification feed conversion rate low, improves breeding effect
Rate, so as to complete the present invention.
In particular it is object of the present invention to provide following aspect:
In a first aspect, providing a kind of SNP marker relevant to pannage conversion ratio, wherein the SNP marker is located at
DHRS4 gene includes subregion, specially No. 7 chromosomes of pig genome version Ensembl Sscrofa 11.1
75245571 bit bases, the SNP marker have A/T polymorphism.
Second aspect provides a kind of acquisition side of SNP marker relevant to pannage conversion ratio described in first aspect
Method, wherein the described method comprises the following steps:
Step 1, pig genomic DNA is obtained；
Step 2, detection obtains SNP site；
Step 3, Genotyping is carried out to SNP site；
Step 4, SNP site and pig growth traits are associated analysis.
The third aspect provides a kind of method identified or auxiliary identifies pannage conversion ratio, wherein the method includes
The step of detecting No. 7 the 75245571st bit base types of chromosome of pig genome version Ensembl Sscrofa 11.1, with
The genotype for determining the gene loci is TT, TA or AA.
Fourth aspect provides the SNP mark that SNP marker described in a kind of first aspect or second aspect the method obtain
Remember the purposes in terms of identifying or assisting identification pannage conversion ratio.
5th aspect provides the SNP mark that SNP marker described in a kind of first aspect or second aspect the method obtain
Remember the application in terms of pig molecule mark assistant breeding.
Beneficial effect possessed by the present invention includes:
(1) SNP marker relevant to pannage conversion ratio provided by the present invention can be used for the early stage breeding of pig, reduce
Breeding cost shortens breeding mid-term, can significantly promote the breeding process of pig；
(2) SNP marker relevant to pannage conversion ratio provided by the present invention, identification accuracy are high, it can be achieved that automatic
Change detection, increases economic efficiency；
(3) SNP marker relevant to pannage conversion ratio provided by the present invention is that the molecular labeling of pig growth traits is auxiliary
Selection is helped to provide scientific basis.
Detailed description of the invention
Fig. 1 shows the primer specificity testing result of the invention with a kind of resulting fluorescent quantitation of preferred embodiment,
In, A shows that the solubility curve of GAPDH reference gene, B show the solubility curve of DHRS4 gene；
Fig. 2 shows a kind of DHRS4 genes of preferred embodiment of the present invention in Landrace different stages of growth skeletal muscle
Expression, wherein A shows that expression of the DHRS4 gene in the chest muscle of Landrace, B show DHRS4 base
Because of the expression in the leg flesh of Landrace, C shows expression of the DHRS4 gene in the dorsal muscles of Landrace；
Fig. 3 shows a kind of location drawing for the SNP site that the screening of preferred embodiment obtains of the present invention, wherein A shows pig
(NCBI Ref. No. is in the site Chr7:75245571SNP of genome version Ensembl Sscrofa 11.1
Rs334250151), B shows the site the Chr7:75245591SNP (NCBI of pig genome version Ensembl Sscrofa 11.1
Ref. No. is rs342446613), C shows the Chr7 of pig genome version Ensembl Sscrofa 11.1:
The site 75253401SNP (NCBI Ref. No. is rs326982309).
Specific embodiment
Below by preferred embodiment and embodiment, the present invention is described in more detail.Illustrated by these, this hair
Bright feature and advantage will become more apparent from clear.
Dedicated word " exemplary " means " being used as example, embodiment or illustrative " herein.Here as " exemplary "
Illustrated any embodiment should not necessarily be construed as preferred or advantageous over other embodiments.
The present inventor is the study found that DHRS4 (Dehydrogenase/Reductase 4, dehydrogenase/reductase enzyme 4) is one
Kind is widely present in the dehydrogenase/reductase enzyme mutually converted between catalysis retinol and retinene in a variety of mammals, there is also
A variety of montage hypotypes, to realize different functional localization and expression.DHRS4 is in vitamin A acid synthesis, steroid metabolism, benzyl
Play an important role in metabolism, and may by influence vitamin A acid synthesis to the Proliferation, Differentiation of cell, cellular signal transduction,
Tumour generates.In addition the enzyme of DHRS4 gene coding and its derivative are also closely related with the reproductive performance of animal, entirely
Retinotic acid and the proliferation of reproduction cell in animal body testis and ovary and differentiation are closely related.
In terms of growth performance, vitamin A acid participates in the myogenic differentiation of Skeletal Muscle Cell, and inducing embryo body further breaks up
For Skeletal Muscle Cell, and improve MyoG (Myogenin, myogenin), (Myogenic factor5, myogenic determine Myf5
The factor), MyHC (Myosin heavy chain genes, myoglobulin heavy chain) etc. the expression of associated transcription factor is generated with flesh
Activity.It can be seen from the above, the growth and development of DHRS4 gene and muscle is closely related, but its molecular signal in muscle development
Conduction path and mechanism still need to further study.Moreover, DHRS4 gene studies focuses primarily upon mouse and human body tumour cell
In, it is less in the developmental research of pig muscle.
Therefore, in the present invention, it is preferred to select DHRS4 gene gene as a purpose, so screen on the gene with feed
The relevant SNP site of conversion ratio.
The first aspect of the present invention provides a kind of SNP marker relevant to pannage conversion ratio, the SNP marker position
In the subregion that includes of DHRS4 gene, the specially Chr7:75245571 of pig genome version Ensembl Sscrofa 11.1
(No. 7 75245571 bit bases of chromosome).
Wherein, the NCBI Ref. No. of the SNP marker is rs334250151, and with A/T polymorphism, the site is corresponding
Three kinds of genotype be TT, TA and AA, TT genotype is the homozygote that the base of the pig SNP site is T, and TA genotype should for pig
The base of SNP site is the heterozygote of T and A, and AA genotype is the homozygote that the base of the pig SNP site is A.
In the present invention, the pig that the SNP marker genotype is AA is TA compared to genotype and the pig of TT is with lower
Feed conversion rate.
A kind of preferred embodiment according to the present invention, using Sequenom MassARRAY technology to SNP marker site
Carry out Genotyping.
Wherein, the Sequenom MassARRAY technology, which refers to, utilizes substance assistant laser desorpted ionized flight time matter
(MALDI-TOF MS) technology of composing, the steps include: PCR amplification target sequence first, and the extension of SNP sequence specific is then added and draws
Object extends 1 base in SNP site.By mass spectrometric vacuum again after the sample analytes of preparation and chip matrix cocrystallization
Pipe is through instantaneous nanosecond (10-9S) light laser excites, and nucleic acid molecules desorption is simultaneously changed into metastable state ion, the flight of electric field intermediate ion
Time is inversely proportional with mass of ion, is obtained by flight time of the time-of-flight detector detection nucleic acid molecules in vacuum tube
The accurate molecular weight of sample analytes, to detect SNP site information.
It can be seen from the above, during carrying out Genotyping to SNP site, extraction, PCR including pig genomic DNA
The step of amplified reaction and single base extension.
In further preferred embodiment, the nucleotide sequence of amplimer P1 and P2 in the pcr amplification reaction
Respectively as shown in SEQ ID NO.1 and SEQ ID NO.2；
The nucleotide sequence of extension primer P3 is as shown in SEQ ID NO.3 in the single base extension.
The present invention also provides a kind of methods of genotyping of SNP site relevant to pannage conversion ratio, the methods
The following steps are included:
(1) genomic DNA of pig is extracted；
(2) genomic DNA for the pig extracted using step (1) is prolonged as template using PCR amplification primer P1, P2 and single base
The extension primer P3 for stretching reaction carries out Sequenom MassARRAY detection, determines pig genome version Ensembl Sscrofa
Genotype at 11.1 No. 7 75245571 bit bases of chromosome.
Wherein, the extracting genome DNA of pig uses method commonly used in the prior art, the nucleotide of primer P1, P2 and P3
Sequence is respectively as shown in SEQ ID NO.1, SEQ ID NO.2 and SEQ ID NO.3.
Another preferred embodiment according to the present invention, the gene of the SNP site relevant to pannage conversion ratio
Parting is measured using direct sequencing or by genotyping kit.
In further preferred embodiment, the genotyping kit includes PCR amplification primer P1, P2 and single alkali
The extension primer P3 of base extension.
In embodiment still more preferably, the genotyping kit further include PCR amplification buffer,
DNTP, PfuDNA polymerase, SNaPshot polymer fluid and PCR product purified reagent,
Wherein, the PCR product purified reagent includes exonuclease, shrimp-alkaline phosphatase and purifying buffer.
A kind of preferred embodiment according to the present invention, the application method of the genotyping kit are as follows: from pig ear or
Genome DNA sample is extracted in its hetero-organization of person；Carry out PCR amplification, purified pcr product；Carry out single base extension；To prolonging
It stretches product and carries out capillary electrophoresis analysis；SNP site analysis, and then obtain genotyping information.
Wherein, SNP site analysis can be used but be not limited to ABI3730XL automatic sequencer.
The second aspect of the present invention provides SNP marker relevant to pannage conversion ratio described in a kind of first aspect
Acquisition methods, the described method comprises the following steps:
Step 1, pig genomic DNA is obtained；
Step 2, detection obtains SNP site；
Step 3, Genotyping is carried out to SNP site；
Step 4, SNP site and pig growth traits are associated analysis.
Optionally, before step 1, the expression to DHRS4 gene in pig different stages of growth skeletal muscle carries out
Detection.
The acquisition methods of the SNP site described further below:
Step 1, pig genomic DNA is obtained.
The present inventor is the study found that the research of DHRS4 gene focuses primarily upon in mouse and human body tumour cell, in pig
The developmental function of Skeletal Muscle Growth rarely has research.Therefore, optionally, before step 1, raw in pig difference to DHRS4 gene
Expression in long stage skeletal muscle is detected.
A kind of preferred embodiment according to the present invention, using the method for quantitative fluorescent PCR to DHRS4 gene in pig difference
Expression in growth phase skeletal muscle is detected, and the detection follows the steps below:
(1) RNA of pig difference skeletal muscle (leg flesh, dorsal muscles, chest muscle) tissue is extracted, and reverse transcription is cDNA；
(2) design carries out the primer of RT-qPCR, selects suitable reference gene, carries out RT-qPCR reaction；
(3) amplification is counted, analyzes expression feelings of the DHRS4 gene in pig different stages of growth skeletal muscle
Condition.
In the present invention, it analyzes after testing, display DHRS4 gene is in the different developments of pig chest muscle, leg flesh and dorsal muscles
Phase expresses, and illustrates the growth and development of skeletal muscle after DHRS4 gene participates in pig birth, is to grow in the subsequent detection gene with pig
The relevant SNP site of character is laid a good foundation, and data supporting is provided.
Wherein, the pig genomic DNA is extracted using method commonly used in the prior art or kit, preferably to certain
The pig genomic DNA of one group extracts.
In the present invention, it is preferred to which the pig genomic DNA to extraction carries out quality testing.
Step 2, detection obtains SNP site.
Wherein, step 2 includes following sub-step:
Step 2-1, design primer are expanded using the pig genomic DNA of acquisition as template, and the DHRS4 gene of pig is obtained
Sequence.
Wherein, referring to the pig DHRS4 genome sequence (NM_214019.2) announced in GeneBank, comprehensively consider primer
Every principle of design utilizes 5.0 software Design primers of Primer permier according to the SNP site information being reported.
In the present invention, it is preferred to design 6 pairs of primers, it is denoted as PA~PF, wherein PAForward and reverse primer sequence respectively such as SEQ
Shown in ID NO.4 and SEQ ID NO.5, PBForward and reverse primer sequence respectively such as SEQ ID NO.6 and SEQ ID NO.7 institute
Show, PCForward and reverse primer sequence respectively as shown in SEQ ID NO.8 and SEQ ID NO.9, PDForward and reverse primer sequence difference
As shown in SEQ ID NO.10 and SEQ ID NO.11, PEForward and reverse primer sequence respectively such as SEQ ID NO.12 and SEQ ID
Shown in NO.13, PFForward and reverse primer sequence respectively as shown in SEQ ID NO.14 and SEQ ID NO.15.
Using pig DNA as template, PCR amplification is carried out using above-mentioned primer, wherein it is mixed to randomly select 100 equivalent sample DNAs
Mixed pond is constructed together, uses 6 sections of primer cDNA clones DHRS4 genes in mixed pond, prepares 1% agarose gel electrophoresis inspection
PCR product clip size and primer specificity are surveyed, the PCR product (5 μ L) and primer (1 μ L) for collecting purpose band carry out two-way survey
Sequence, direct Sequencing peak figure clean background is readable and includes set peak, and obtains the SNP of heterozygosity >=0.10.
In the present invention, it is preferred to carry out Sanger sequencing analysis to pcr amplification product.
Step 2-2 carries out sequence alignment, obtains SNP site.
In the present invention, it is preferred to the pig that will be delivered on sequencing result and GeneBank by software Chromas Pro software
DHRS4 gene order is compared, and obtains SNP site.
Step 3, Genotyping is carried out to SNP site.
Wherein, it is preferred to use Sequenom MassARRAY technology carries out Genotyping to SNP site.
Step 4, the genotype data of SNP site and pig growth traits are associated analysis.
Wherein, step 4 includes following sub-step:
Step 4-1 obtains the phenotypic data of pig growth traits.
In the present invention, the growth traits of the pig includes average daily gain, feed conversion rate and the thickness of backfat, wherein
Age in days is surveyed by the beginning, knot surveys age in days, beginning survey weight (g), knot surveys weight (g), knot survey feed consumption (g) is calculated and averagely increased day by day
Weight, feed conversion rate, formula are as follows:
Average daily gain (g)=[knot surveys weight (g)-and begins to survey weight (g)]/[knot surveys age in days-beginning and surveys age in days]；
Feed conversion rate (%)=knot surveys feed consumption (g)/[knot surveys weight (g)-and begins to survey weight (g)]；
The measurement of the thickness of backfat: the back-fat thickness of test swinery individual is measured using praetersonic back fat analyzer.Starting
When measurement, allows naturally quiet stand of pig to remain stationary, in the cropping at the upward 5cm of dorsal line at last root rib cage of pig, coat coupling
Back fat instrument probe is placed on this position by mixture, is recorded after instrument readings are stablized；Continuous measurement 3 times, is averaged, and record number
According to.
The genotype data of SNP site and pig growth traits are associated analysis, acquisition and pannage by step 4-2
The relevant SNP marker of conversion ratio.
In the present invention, it is preferred to calculate the genotype frequency and gene frequency of SNP site using PopGene3.2, so
Afterwards using the relevance of GLM analysis SNP site and pig growth traits in SAS9.2 software, obtain and the significant phase of pannage conversion ratio
The SNP marker of pass.Simultaneously, additionally it is possible to obtain the low genotype of pannage conversion ratio.
The third aspect of the present invention provides a kind of method identified or auxiliary identifies pannage conversion ratio, the method
Include the steps that detecting the base type at the Chr7:75245571 of pig genome version Ensembl Sscrofa11.1, with true
The genotype of the fixed gene loci is TT, TA or AA.
Wherein, the height of pannage conversion ratio is determined according to the genotype for being detected as gene loci, genotype is the pig of AA
There is lower feed conversion rate compared to the pig that genotype is TA and TT.
The fourth aspect of the present invention, provides SNP marker described in a kind of first aspect or second aspect the method obtains
SNP marker identify or assist identification pannage conversion ratio in terms of purposes.
The fifth aspect of the present invention provides SNP site described in a kind of first aspect in pig molecule mark assistant breeding side
The application in face, it is described application the following steps are included:
Step I extracts the genomic DNA of pig to be measured；
Step II detects the genotype of pig to be measured by Sequenom MassARRAY technology；
Step III carries out the breeding of the low pig Dominant variety of feed conversion rate according to genotype.
Wherein, in step II, the amplimer that is used in Sequenom MassARRAY technology detection process to for P1 and
P2, the extension primer used is P3, and the primer sequence is respectively such as SEQ ID NO.1, SEQ ID NO.2 and SEQ ID NO.3
It is shown.
Embodiment
The present invention is further described below by way of specific example, but these examples are only exemplary, not to this
The protection scope of invention constitutes any restrictions.
The experimental animal applied in embodiment:
Test acquires the dorsal muscles, chest muscle, leg flesh of 30 ages in days (30d), 180 ages in days (180d), 300 ages in days (300d) Landrace
Sample is organized, each period acquires 3 and repeats as biology, sample is put into cryopreservation tube, puts into liquid nitrogen rapidly, is transported to experiment
Room is put into -80 DEG C of refrigerators and saves backup.
The pig ear sample for acquiring 235 healthy adult Lan Sibai pigs, is put into 75% alcohol, -20 DEG C of preservations.It arranges simultaneously
Its Product archives, including begin survey age in days (day), knot survey age in days (day), begin survey weight (g), knot survey weight (g), knot survey feed consumption (g),
The performance indicators such as the thickness of backfat (mm).
Test sample is acquired from Shandong Lan Sizhong industry limited liability company, and swine rearing process and experiment in this research
Operating method all follows the requirement of the raising and operation of experimental animal.Wherein, Lan Sibai pig is that an indigo plant think of kind industry share in Shandong has
Limit company and Institute of Animal Sciences, Chinese Academy of Agricultural Sciences are black with English system Large White and domestic excellent local varieties Yimeng
Pig is parent, the stronger resistance of the black pig in Yimeng and the excellent growth performance characteristic of English system Large White is made full use of, by 6 generation
The Systematic Breeding recorded, last 17 years, what directive breeding came out has the fast speed of growth, adaptable, reproductive performance and succulence
The stable specialized new population of energy, contains black 12.5% blood relationship of pig in Yimeng.
The reagent source and instrument and equipment of embodiment application:
Reagent: (1) DNA extraction kit (animal tissue DNA extraction): TIANGEN, Tiangeng biochemical technology (Beijing) are limited
Company；(2) chloroform (Beijing chemical reagents corporation), dehydrated alcohol (Beijing chemical reagents corporation), isopropanol (Beijing chemical reagent
Company)；(3) RNAiso Plus:TaKaRa, Japan；(4) SYBR Primer ScriptTM RT-PCR Kit:TaKaRa, day
This；(5) SYBR Premix EX Taq:Tli RNaseH Plus, TaKaRa, Japan.
Instrument and equipment: (1) full automatic gel imaging system: Bio-rad company；(2) electrophoresis tank: Bio-rad company；(3)
Nucleic acid-protein concentration mensuration instrument (Nano-100): Hangzhou Ao Sheng company；(4) PCR instrument (C1000Touch TM): Bio-Rad is public
Department；(5) fluorescence quantitative PCR instrument (Applied Biosystems 7500, ABI 7500)；(6) centrifuge (Centrifuge
5810R): Eppendorf company.
The acquisition of 1 SNP marker of embodiment
1, expression of the DHRS4 gene in pig different stages of growth skeletal muscle is detected.
1.1, the extraction and reverse transcription of RNA
Take Landrace 30d, 180d and 300d leg flesh, dorsal muscles and breast muscle (every group of 3 biology repeats, totally 27
A sample), total serum IgE is extracted according to total RNA from animal tissues extracts kit specification, it is dense with nucleic acid-protein concentration meter measurement RNA
Degree and OD (optical density, optical density) value, qualified RNA sample is referring to RevertAid First Strand cDNA
Synthesis Kit reverse transcription reagent box specification synthesizes the first chain cDNA and takes 2000ng's according to the total rna concentration of measurement
Random primer Oligo (dT) 1 μ L is added in RNA solution, and is mended volume to 12 μ L, after oscillation mixes, 70 DEG C of heating 5 with distilled water
Minute, it takes out and places 3-5 minutes on ice, 5 × RT Buffer, 4 μ L, 2 dNTP μ L, RiboLock are added after brief centrifugationTM1μ
L、RevertAidTM1 μ L of M-MLV, 20 μ L of total system, is put into PCR instrument after mixing, 42 DEG C of program of setting heating 1 hour, and 70
DEG C, 10min；4 DEG C, ∞.- 20 DEG C of refrigerators are put into after the reaction was completed to save backup.
1.2, real-time fluorescence quantitative PCR (RT-qPCR)
It the use of GAPDH is reference gene, with Permier primer5.0 software design RT-qPCR primer and internal control primer,
The RT-qPCR primer pair is P4 and P5, and nucleotide sequence is respectively as shown in SEQ ID NO.16 and SEQ ID NO.17, internal reference
Primer pair is P6 and P7, nucleotide sequence respectively as shown in SEQ ID NO.18 and SEQ ID NO.19, RT-qPCR react according to
SYBR Premix Ex TaqTM kit (TaKaRa) specification carries out, and carries out in ABI prism 7500PCR system, instead
Answering total system is 15 μ L:2 × SYBR Premix EX Taq, 7.2 μ L, II 0.3 μ L of ROXReference Dye, Primer
0.3 μ L of sense, 0.8 μ L of cDNA template, 6.1 ddH2O μ L, reaction condition are 95 DEG C of 5min, 95 DEG C of 5s, 60 DEG C of 1min, circulation
Solubility curve analysis is carried out after 40 times, DHRS4 gene and GAPDH reference gene carry out under same reaction condition, each sample
It is repeated using 3 technologies, is finally averaged.
Wherein, the primer specificity testing result of the fluorescent quantitation is as shown in Figure 1, DHRS4 gene is different in Landrace
Expression in growth phase skeletal muscle is as shown in Figure 2.
A in Fig. 1 shows that the solubility curve of GAPDH reference gene, the B in Fig. 1 show the molten of DHRS4 gene
Solution curve, wherein carry out specific amplification using 27 muscle tissue samples of the primer pair Landrace, obtain corresponding 27
Amplification curve.
As shown in Figure 1, the solubility curve (27 curves of 27 samples) of GAPDH reference gene is single peak type,
Without other miscellaneous peaks；The solubility curve (27 curves of 27 samples) of DHRS4 gene is also single peak type, without other miscellaneous peaks.
Illustrate that the amplified production of above-mentioned two pairs of primers is single, be specific product, is generated without non-specific amplifications such as primer dimers,
Show the height of RT-qPCR primer specificity used by the present embodiment, can be used for the subsequent expression inspection of target gene DHRS4 gene
It surveys.
A, B and C in Fig. 2 respectively illustrate expression feelings of the DHRS4 gene in the chest muscle, leg flesh and dorsal muscles of Landrace
Condition, as shown in Figure 2, DHRS4 gene have expression in the different development stage of Landrace chest muscle, leg flesh and dorsal muscles, and in 30d
When expression quantity highest.The expression pattern of DHRS4 gene is close in chest muscle and leg flesh, and expression is remarkably decreased when 180d and 300d；
The expression quantity of DHRS4 gene does not change significantly with the increase of age in days in dorsal muscles.
The meat yield and meat quality of pig are closely related with skeletal development process, and skeletal development includes that embryonic period, embryonic phase flesh is fine
The development of muscle fibre and the regeneration of manhood skeletal muscle after the formation of dimension, birth, the quantity of muscle fibre is no longer after birth
Variation is understood some slow switch fibers and is gradated as fast muscle fiber, the order of conversion is slow second is that over time
Flesh, fast muscle, osculant fast muscle and fast white muscle show the variation of muscle fibre hypertrophy in form.Show that DHRS4 gene exists in Fig. 2
Gene expression is higher in Landrace 30d musculature, as the growth expression of muscle sharply declines, thus it is speculated that DHRS4 gene
The growth course for participating in Animal muscles, in the process of muscle fiber types conversion, i.e., during slow muscle is converted to fast muscle,
The expression quantity of DHRS4 gene is on a declining curve.
It is therefore contemplated that DHRS4 gene takes part in the growth and development of skeletal muscle after pig birth.
2, the extraction and quality testing of pig genomic DNA
(1) the pig ear tissue of clip 30mg size, shreds and is placed into the centrifuge tube of 1.5ml；
(2) take 50ml centrifuge tube, the Proteinase K of final concentration of 0.4mg/ml and lysis buffer are carried out it is uniformly mixed,
0.5ml lysate is added into the centrifuge tube of the 1.5ml equipped with pig ear tissue to be cracked；
(3) centrifuge tube is uniformly placed on constant-temperature table (tight tube sealing lid, in order to avoid intravenous extravasation), and 55 DEG C stand overnight,
Digestion to visually observe less than tissue sample；
(4) after sample sufficiently cracks, centrifuge tube is taken out, 0.3ml saturated sodium chloride solution is added in every pipe, runs
It is sufficiently mixed for several times, is then put on ice, ice bath 15min；
(5) after ice bath, 12000rpm room temperature is centrifuged 15min, and supernatant is slowly carefully transferred to new 1.5ml and is centrifuged
Guan Zhong is carefully avoided to precipitate and be poured out together；
(6) be added it is reverse with the isometric isopropanol of supernatant until have in solution flocculent deposit occur (if without flocculent deposit,
- 20 DEG C of refrigerators of solution can be placed 2h or 4 DEG C of refrigerator to stand overnight)；
(7) 12000rpm room temperature is centrifuged 15min, removes supernatant fluid and (notices the white of observation centrifugation bottom of the tube during this
Color DNA precipitating, is sure not to outwell it together with supernatant)；
(8) 0.5ml 70% ethyl alcohol is added in each centrifuge tube, gently overturns sufficiently to rinse above-mentioned be precipitated out
DNA；
(9) 10000rmp is centrifuged 30s, sops up the ethyl alcohol in centrifuge tube with liquid-transfering gun, the DNA of precipitating is stayed in pipe
(taking care not siphon away the DNA precipitating in pipe, replacement can not be had to when pipette tips used in this step operate between each centrifuge tube)；
(10) the DNA natural air drying of extraction is dried into 10min；
(11) liquid-transfering gun takes the TE buffer of 0.1ml in each pipe, and above-mentioned DNA precipitating is re-dissolved, sets it in 55 DEG C
It places 2 hours, timing shaken several times, so that DNA sufficiently dissolves；
(12) after completely dissolution to DNA, 1 μ LDNA is drawn, detects DNA concentration and OD260/ with ultramicrospectrophotometer
OD280 value shows there is RNA pollution if OD260/OD280 value is greater than 1.9；If OD260/OD280 value, less than 1.6, showing can
Can there are phenol or protein contamination；OD260/OD280 value ≈ 1.8, as pure dna；
Meanwhile mentioned DNA mass, single band, then it represents that DNA satisfactory quality are detected with agarose gel electrophoresis.
3, SNP site is detected
Referring to the pig DHRS4 genome sequence (NM_214019.2) announced in GeneBank, design of primers is comprehensively considered
Every principle is denoted as P with 5.0 software design 6 of Primer permier to primer according to the SNP site information being reportedA
~PF, wherein PAForward and reverse primer sequence respectively as shown in SEQ ID NO.4 and SEQ ID NO.5, PBForward and reverse primer
Sequence is respectively as shown in SEQ ID NO.6 and SEQ ID NO.7, PCForward and reverse primer sequence respectively such as SEQ ID NO.8 and
Shown in SEQ ID NO.9, PDForward and reverse primer sequence respectively as shown in SEQ ID NO.10 and SEQ ID NO.11, PEJust
Reverse primer sequences are respectively as shown in SEQ ID NO.12 and SEQ ID NO.13, PFForward and reverse primer sequence respectively such as SEQ
Shown in ID NO.14 and SEQ ID NO.15.
Using pig DNA as template, PCR amplification is carried out using above-mentioned primer, wherein it is mixed to randomly select 100 equivalent sample DNAs
Mixed pond is constructed together, uses 6 sections of primer cDNA clones DHRS4 genes in mixed pond, prepares 1% agarose gel electrophoresis inspection
PCR product clip size and primer specificity are surveyed, the PCR product (5 μ L) and primer (1 μ L) for collecting purpose band carry out two-way survey
Sequence, direct Sequencing peak figure clean background is readable and includes set peak, and obtains the SNP of heterozygosity >=0.10.
Sanger sequencing is carried out to above-mentioned PCR product, the sequencing result of acquisition passes through DNAMAN and Chromas Pro software
It is compared with the pig DHRS4 gene order delivered on GeneBank, filters out 3 sites SNPs altogether, as a result as shown in Figure 3.
A in Fig. 3 shows the site Chr7:75245571SNP of pig genome version Ensembl Sscrofa 11.1
(NCBI Ref. No. is rs334250151)；
B in Fig. 3 shows the site Chr7:75245591SNP of pig genome version Ensembl Sscrofa 11.1
(NCBI Ref. No. is rs342446613)；
C in Fig. 3 shows the site Chr7:75253401SNP of pig genome version Ensembl Sscrofa 11.1
(NCBI Ref. No. is rs326982309).
From the figure 3, it may be seen that 3 SNP sites filtered out are located at pig genome version Ensembl Sscrofa 11.1
No. 7 chromosomes 75245571,75245591 and 75253401 bit bases go out, Genetic elements annotation shows that 3 SNP sites are located
Subregion is included in DHRS4 gene.
4, SNP site Genotyping and Quality Control
According to the SNP site context information design PCR amplification primer and base extension primer detected, wherein
The PCR amplification primer nucleotide sequences P1 and P2 of rs334250151 as shown in SEQ ID NO.1 and SEQ ID NO.2, prolong by base
Object P3 extend as shown in SEQ ID NO.3；PCR amplification primer nucleotide sequences P8 and the P9 such as SEQ ID of rs342446613
Shown in NO.20 and SEQ ID NO.21, base extension primer P10 is as shown in SEQ ID NO.22；The PCR of rs326982309 expands
Increase primer nucleotide sequences P11 and P12 as shown in SEQ ID NO.23 and SEQ ID NO.24, base extension primer P13 such as SEQ
Shown in ID NO.25.
Pcr amplification reaction is carried out by template of the DNA sample of 235 Lan Sibai pigs, and PCR product is purified, is protected
The accuracy that card base extends is existed then with highly sensitive mass spectrum chip cocrystallization using Sequenom MassARRAY technology
Group carries out Genotyping to the SNP site on DHRS4 gene.
Wherein, the genotyping result is as shown in table 1, wherein the genotype frequency of SNP site is calculated using PopGene3.2
And gene frequency:
Table 1
As shown in Table 1, in rs334250151 site primer to three genotype: TT, TA and AA, and TT type is advantage
Genotype, genotype frequency (0.668) are higher than the frequency of TA (0.302) and AA genotype (0.030)；In rs342446613
Site primer is to three kinds of genotype: TT, TC and CC, wherein the frequency (0.494) of TC genotype be higher than TT genotype (0.327) and
The frequency of CC genotype (0.179)；In rs326982309 site primer to three genotype: AA, TA and TT, AA genotype frequency
Rate (0.698) is higher than the frequency of TA genotype (0.285) and TT genotype (0.017).
5, the correlation of above-mentioned 3 SNP sites and pig growth traits is analyzed
5.1 obtain the growth traits phenotypic data of pig
Age in days is surveyed by the beginning, knot surveys age in days, beginning survey weight (g), knot surveys weight (g), knot survey feed consumption (g) is calculated and averagely increased day by day
Weight, feed conversion rate, formula are as follows:
Average daily gain (g)=[knot surveys weight (g)-and begins to survey weight (g)]/[knot surveys age in days-beginning and surveys age in days]
Feed conversion rate (%)=knot surveys feed consumption (g)/[knot surveys weight (g)-and begins to survey weight (g)]
The measurement of the thickness of backfat: the back-fat thickness of test swinery individual is measured using praetersonic back fat analyzer.Starting
When measurement, allows naturally quiet stand of pig to remain stationary, in the cropping at the upward 5cm of dorsal line at last root rib cage of pig, coat coupling
Back fat instrument probe is placed on this position by mixture, is recorded after instrument readings are stablized.Continuous measurement 3 times, is averaged, and record number
According to.Then for statistical analysis to above-mentioned phenotypic data with Excel (Microsoft Office 2017).
5.2 association analysis analysis
Using the genotype of the GLM process analysis procedure analysis SNP site in SAS9.2 software and the relevance of pig growth traits, adopted
Fixed-effect model is as follows:
Y=μ+gi+mk+e；
Wherein, y is the phenotypic record of growth traits；μ is the overall average of character；Gi is genotype effects；Mk is production
Month effect；E is random error.
Data indicate that P value < 0.05 indicates there is significant difference with probability value and mean+SD.
The results are shown in Table 2 for association analysis:
Table 2
Wherein, data are indicated with mean ± standard deviation, and shoulder mark lowercase difference indicates significant difference (p < 0.05).
As shown in Table 2, the existing site rs334250151 is significant related (P < 0.05) to feed conversion rate, AA genotype individuals
Feed conversion rate (2.24) be substantially less than TA type (2.40) and TT type (2.45), it is not significant with average daily gain and the thickness of backfat
It is related.And the site rs342446613, rs326982309 and average daily gain, feed conversion rate, average backfat thickness etc. are without aobvious
The association of work.
In summary, the polymorphic site rs334250151 on DHRS4 gene be expected as with Lan Sibai pannage conversion ratio
The relevant candidate genetic marker site of character, for instructing the breeding of Lan Sibai pig meat-producing traits to work.Wherein it is possible to by true
The base type in the site pig DHRS4 gene rs334250151 is determined to determine the genotype of pig individual, to assist identification pannage
The individual of conversion ratio, AA genotype has lower feed conversion rate.
It is described the invention in detail above in conjunction with detailed description and exemplary example, but these explanations are simultaneously
It is not considered as limiting the invention.It will be appreciated by those skilled in the art that without departing from the spirit and scope of the invention,
Can be with various equivalent substitutions, modifications or improvements are made to the technical scheme of the invention and its embodiments, these each fall within the present invention
In the range of.
SEQUENCE LISTING
<110>source Tianjin Nuo Hezhi biological information Science and Technology Ltd.
<120>molecular labeling relevant to pannage conversion ratio, identification and its application
<130> 2019
<160> 25
<170> PatentIn version 3.5
<210> 1
<211> 30
<212> DNA
<213>amplimer P1(artificial sequence)
<400> 1
acgttggatg gtgaaagtag cagagaaggg 30
<210> 2
<211> 30
<212> DNA
<213>amplimer P2(artificial sequence)
<400> 2
acgttggatg tgtacttttt ccagaggcgg 30
<210> 3
<211> 24
<212> DNA
<213>extension primer P3(artificial sequence)
<400> 3
ccttttcctc cgttggagcc tacc 24
<210> 4
<211> 20
<212> DNA
<213>forward primer (artificial sequence) of amplimer PA
<400> 4
tccctccttg gctatctgct 20
<210> 5
<211> 20
<212> DNA
<213>reverse primer (artificial sequence) of amplimer PA
<400> 5
agccaggact agtctccctg 20
<210> 6
<211> 20
<212> DNA
<213>forward primer (artificial sequence) of amplimer PB
<400> 6
gagtgtgtgc tgtcttcgga 20
<210> 7
<211> 20
<212> DNA
<213>reverse primer (artificial sequence) of amplimer PB
<400> 7
gccttgaatc ttccctgcct 20
<210> 8
<211> 20
<212> DNA
<213>forward primer (artificial sequence) of amplimer PC
<400> 8
agagagtcag ggagccagag 20
<210> 9
<211> 20
<212> DNA
<213>reverse primer (artificial sequence) of amplimer PC
<400> 9
gaccctaagg ggcatttgct 20
<210> 10
<211> 20
<212> DNA
<213>forward primer (artificial sequence) of amplimer PD
<400> 10
ggtcaagagt tgctggctct 20
<210> 11
<211> 20
<212> DNA
<213>reverse primer (artificial sequence) of amplimer PD
<400> 11
agcagatagc caaggaggga 20
<210> 12
<211> 20
<212> DNA
<213>forward primer (artificial sequence) of amplimer PE
<400> 12
ctgatgggga aatgcctggt 20
<210> 13
<211> 20
<212> DNA
<213>reverse primer (artificial sequence) of amplimer PE
<400> 13
cctgtgacca tgggaacctc 20
<210> 14
<211> 20
<212> DNA
<213>forward primer (artificial sequence) of amplimer PF
<400> 14
gccaggcagt tcaccctaat 20
<210> 15
<211> 20
<212> DNA
<213>reverse primer (artificial sequence) of amplimer PF
<400> 15
aagtgctgga acaaccccaa 20
<210> 16
<211> 20
<212> DNA
<213>RT-qPCR amplimer P4(artificial sequence)
<400> 16
gccgtcaacc cattctttgg 20
<210> 17
<211> 20
<212> DNA
<213>RT-qPCR amplimer P5(artificial sequence)
<400> 17
gcaccactgc ctttgtcatc 20
<210> 18
<211> 20
<212> DNA
<213>reference gene amplimer P6(artificial sequence)
<400> 18
agggcatcct gggctacact 20
<210> 19
<211> 20
<212> DNA
<213>reference gene amplimer P7(artificial sequence)
<400> 19
tccaccaccc tgttgctgta 20
<210> 20
<211> 30
<212> DNA
<213>amplimer P8(artificial sequence)
<400> 20
acgttggatg gtgaaagtag cagagaaggg 30
<210> 21
<211> 30
<212> DNA
<213>amplimer P9(artificial sequence)
<400> 21
acgttggatg tgtacttttt ccagaggcgg 30
<210> 22
<211> 16
<212> DNA
<213>extension primer P10(artificial sequence)
<400> 22
gctcagtgtt gatcgt 16
<210> 23
<211> 30
<212> DNA
<213>amplimer P11(artificial sequence)
<400> 23
acgttggatg aggtggttga aacatggtgg 30
<210> 24
<211> 30
<212> DNA
<213>amplimer P12(artificial sequence)
<400> 24
acgttggatg tgggtattcc cactatgagc 30
<210> 25
<211> 24
<212> DNA
<213>extension primer P13(artificial sequence)
<400> 25
aactgtttat tcatctttta aagt 24
Claims (9)
1. a kind of SNP marker relevant to pannage conversion ratio, which is characterized in that the SNP marker is located at the interior of DHRS4 gene
Containing subregion, No. 7 the 75245571st bit bases of chromosome of specially pig genome version Ensembl Sscrofa11.1, institute
SNP marker is stated with A/T polymorphism.
2. the SNP marker according to requiring 1, which is characterized in that the corresponding three kinds of genotype of the SNP marker be respectively TT,
TA and AA,
The pig that genotype is AA is TA compared to genotype and the pig of TT has lower feed conversion rate.
3. the SNP marker according to requiring 1, which is characterized in that SNP marker site carry out Genotyping method include
The step of pcr amplification reaction and single base extension,
Preferably, in the pcr amplification reaction nucleotide sequence of amplimer P1 and P2 respectively such as SEQ ID NO.1 and SEQ
Shown in ID NO.2；
The nucleotide sequence of extension primer P3 is as shown in SEQ ID NO.3 in the single base extension.
4. the acquisition methods of SNP marker relevant to pannage conversion ratio, feature described in a kind of one of claims 1 to 3
It is, the described method comprises the following steps:
Step 1, pig genomic DNA is obtained；
Step 2, detection obtains SNP site；
Step 3, Genotyping is carried out to SNP site；
Step 4, SNP site and pig growth traits are associated analysis.
5. according to the method described in claim 4, it is characterized in that, optionally, before step 1, to DHRS4 gene pig not
It is detected with the expression in growth phase skeletal muscle.
6. according to the method described in claim 4, it is characterized in that, step 4 includes following sub-step:
Step 4-1 obtains the phenotypic data of pig growth traits；
The genotype data of SNP site and pig growth traits are associated analysis, obtain and convert with pannage by step 4-2
The relevant SNP marker of rate.
7. a kind of method of identification or auxiliary identification pannage conversion ratio, which is characterized in that the method includes detecting pig gene
The step of No. 7 the 75245571st bit base types of chromosome of group version Ensembl Sscrofa11.1, to determine the gene position
The genotype of point is TT, TA or AA.
8. the SNP mark that one of SNP marker or claim 4 to 6 the method described in a kind of one of claims 1 to 3 obtain
Remember the purposes in terms of identifying or assisting identification pannage conversion ratio.
9. the SNP mark that one of SNP marker or claim 4 to 6 the method described in a kind of one of claims 1 to 3 obtain
Remember the application in terms of pig molecule mark assistant breeding.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910630651.1A CN110317880B (en) | 2019-07-12 | 2019-07-12 | Molecular marker related to pig feed conversion rate, identification and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910630651.1A CN110317880B (en) | 2019-07-12 | 2019-07-12 | Molecular marker related to pig feed conversion rate, identification and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110317880A true CN110317880A (en) | 2019-10-11 |
| CN110317880B CN110317880B (en) | 2022-11-15 |
Family
ID=68122066
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910630651.1A Active CN110317880B (en) | 2019-07-12 | 2019-07-12 | Molecular marker related to pig feed conversion rate, identification and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110317880B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111500746A (en) * | 2020-05-22 | 2020-08-07 | 华中农业大学 | SNP molecular marker related to feed conversion efficiency of pigs |
| CN112094924A (en) * | 2020-10-21 | 2020-12-18 | 海南罗牛山种猪育种有限公司 | Genetic marker taking MYST2 gene as lean meat percentage and average daily gain of large and white pig, detection method and application thereof |
| CN113699246A (en) * | 2021-07-26 | 2021-11-26 | 华南农业大学 | SNP molecular marker influencing pig feed conversion efficiency traits and application thereof |
| CN114182025A (en) * | 2021-12-14 | 2022-03-15 | 华中农业大学 | SNP molecular marker related to pig feed conversion rate and application thereof |
| CN114250307A (en) * | 2020-09-25 | 2022-03-29 | 中国农业科学院农业基因组研究所 | Molecular marker for evaluating day age of pigs with weight of 100kg and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107937556A (en) * | 2017-11-14 | 2018-04-20 | 中国农业大学 | One and the relevant SNP site of pannage conversion ratio and its application |
| CN109402270A (en) * | 2018-12-07 | 2019-03-01 | 佛山科学技术学院 | One kind SNP marker relevant to Large White growth traits and its application |
-
2019
- 2019-07-12 CN CN201910630651.1A patent/CN110317880B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107937556A (en) * | 2017-11-14 | 2018-04-20 | 中国农业大学 | One and the relevant SNP site of pannage conversion ratio and its application |
| CN109402270A (en) * | 2018-12-07 | 2019-03-01 | 佛山科学技术学院 | One kind SNP marker relevant to Large White growth traits and its application |
Non-Patent Citations (1)
| Title |
|---|
| 成宏: "猪4个候选基因SNP检测及与饲料转化效率性状的关联性分析", 《中国优秀博硕士学位论文全文数据库(硕士)》 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111500746A (en) * | 2020-05-22 | 2020-08-07 | 华中农业大学 | SNP molecular marker related to feed conversion efficiency of pigs |
| CN114250307A (en) * | 2020-09-25 | 2022-03-29 | 中国农业科学院农业基因组研究所 | Molecular marker for evaluating day age of pigs with weight of 100kg and application thereof |
| CN114250307B (en) * | 2020-09-25 | 2024-05-14 | 中国农业科学院农业基因组研究所 | Molecular marker for evaluating age of pigs reaching 100kg body weight and application thereof |
| CN112094924A (en) * | 2020-10-21 | 2020-12-18 | 海南罗牛山种猪育种有限公司 | Genetic marker taking MYST2 gene as lean meat percentage and average daily gain of large and white pig, detection method and application thereof |
| CN113699246A (en) * | 2021-07-26 | 2021-11-26 | 华南农业大学 | SNP molecular marker influencing pig feed conversion efficiency traits and application thereof |
| CN113699246B (en) * | 2021-07-26 | 2023-07-11 | 华南农业大学 | SNP molecular marker affecting pig feed conversion efficiency character and application thereof |
| CN114182025A (en) * | 2021-12-14 | 2022-03-15 | 华中农业大学 | SNP molecular marker related to pig feed conversion rate and application thereof |
| CN114182025B (en) * | 2021-12-14 | 2023-03-14 | 华中农业大学 | SNP molecular marker related to pig feed conversion rate and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110317880B (en) | 2022-11-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110317880A (en) | Molecular labeling relevant to pannage conversion ratio, identification and its application | |
| CN101629209B (en) | Method for detecting cattle Six6 gene single nucleotide polymorphism | |
| CN109929935B (en) | Method for identifying pig backfat thickness based on rs80995809 locus genotyping and application thereof | |
| CN104561367B (en) | A kind of SNP for influencing label of pig fat deposition description character and its application | |
| CN108728552B (en) | It is a kind of influence duroc eye muscle area character molecular labeling and application | |
| CN105969845B (en) | The molecular labeling of eye muscle area character related gene SVEP1 and its application | |
| CN105063021B (en) | The SNP marker related to label of pig fat deposition description and its application | |
| CN103290123B (en) | Detecting method and kit of cattle IGF2 (Insulin-like Growth Factor 2) gene mononucleotide polymorphism | |
| CN112195253B (en) | 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 | |
| CN108949907A (en) | One kind SNP marker primer pair relevant to Suhuai pig intramuscular fat content and its application | |
| CN109811063A (en) | One kind SNP marker relevant to growth speed of pigs and its application | |
| CN109402270A (en) | One kind SNP marker relevant to Large White growth traits and its application | |
| CN109234404A (en) | A kind of method and primer special for identifying sheep meat-producing traits | |
| CN111926086A (en) | Molecular marker influencing oblique growth of chicken body and application thereof | |
| CN114921568A (en) | SNP molecular marker related to Qinchuan cattle body ruler and meat quality traits and application thereof | |
| CN101921848B (en) | Method for detecting single nucleotide polymorphism (SNP) of cattle MGAT2 gene | |
| CN101921852B (en) | Method for detecting single nucleotide polymorphism of cattle AdPLA gene | |
| CN101671726B (en) | Method for detecting single nucleotide polymorphism (SNP) of ox PRDM16 gene | |
| CN104498610B (en) | RFLP method and kit for detecting SNP locus of MEF2C gene of cattle | |
| CN102776184B (en) | Genetic marker using pig CKM (creatine kinase muscle) 5' flanking promoter region SNP (single nucleotide polymorphism) as pig carcass traits and application | |
| CN109055578A (en) | A kind of PLAG1 gene SNP label auxiliary quickly detects the method and its application of ox growth traits | |
| CN113699247B (en) | SNP molecular marker related to pig residual feed intake on pig chromosome 1 and application thereof | |
| CN104762373A (en) | Chicken muscle fiber-type property-related gene molecule marker and use thereof | |
| CN109022594A (en) | A kind of ox AHSG gene SNP marker related with cattle feed transformation efficiency | |
| CN107130037B (en) | Method and special kit for auxiliary detection of cattle growth and carcass traits through TNNI1 gene |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |