CN107586854B - It is a kind of influence duroc waistline character molecular labeling and application - Google Patents
It is a kind of influence duroc waistline character molecular labeling and application Download PDFInfo
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Abstract
The invention belongs to molecular marker assisted selection and Animal Genetics technical fields, and in particular to it is a kind of influence duroc waistline character molecular labeling and application.The SNP site of the molecular labeling of the influence duroc waistline character corresponds to the 3400805th nucleotide site T > C mutation on international pig genome No. 4 chromosomes of 10.2 version reference sequences.The present invention also provides a kind of for identifying that above-mentioned influence Pigs Kidney encloses the primer pair of the molecular labeling of character, pass through above-mentioned molecular labeling and primer pair, the molecular mark technology of efficiently and accurately can be established, it is applied in the improvement of Duroc boar waistline character inheritance, to improve the waistline of offspring pig, and then meat yield is improved, enterprise profit is improved, core competitiveness is increased.
Description
Technical field
The invention belongs to molecular marker assisted selection and Animal Genetics technical fields, and in particular to a kind of influence Du Luo
Gram Pigs Kidney encloses molecular labeling and the application of character.
Background technique
Under market environment based on mainly being evaluated in Chinese pork pig with live hog, the selection of figure phenotype is particularly important.
External boar market can seldom judge its value according to figure, nevertheless, still can forelimb, hind leg, movement to pig
The appearance traits such as state, posture, back, linea ventralis carry out comprehensive assessment selection.Most of China live pig in the market, linea ventralis (abdominal circumference,
Waistline) value that determines pork pig, directly affect the profit on sales of end product.Since abdominal circumference is big, waistline is small, linea ventralis is uneven
Directly, it is commonly called as pregnant, potbelly, this boar causes pork pig finishing loss larger, affect boar and finally sell since fat meat is more
Profit out.In addition, being unwilling to introduce a fine variety so that such boar utilization rate drops so as to cause commodity pig farm since kind of pork tripe sirloin is larger
Low situation is commonplace, and for pig raising, enterprise is had brought tremendous economic losses.Because the selection of figure need to take into account it is numerous, very
Difficulty takes into account waistline while selecting other characters, brings difficulty for waistline improvement.The Phenotypic Selection of waistline and measurement are not
There are system and in detail research.By domestic market survey, requirement of the different zones to waistline is different, this causes to select in boar
When educating, the breeding objective of the appearance traits such as each terminal male strain waistline slightly has difference.For this purpose, carrying out for market demands to waist
It encloses phenotype and carries out more detailed measurement and selection, furthermore it is also possible to which the analysis by this phenotype test and genome is reflected
Not Kong Zhi boar figure gene and major gene loci, using the means of molecular breeding fundamentally solve waistline improve it is difficult
Topic.
Duroc is one of bacon hogs kind famous in the world.In the breeding of method of modern goods pork pig, Du Luo
It gram is used generally as terminal male, coverage is larger.Therefore, the waistline character of duroc is improved, Ke Yiyou
The figure of effect ground improvement market pig, improves its selling price.
Summary of the invention
In order to overcome deficiency and disadvantage existing for boar figure breeding in the prior art, the primary purpose of the present invention is that mentioning
For a kind of molecular labeling for influencing duroc waistline character.
Another object of the present invention is to provide the applications of the molecular labeling of above-mentioned influence duroc waistline character.
A further object of the present invention is to provide a kind of molecular labelings for identifying above-mentioned influence duroc waistline character
Primer pair.
Fourth object of the present invention is to provide the application of above-mentioned primer pair.
Of the invention the 5th is designed to provide a kind of genetic improvement method of duroc.
A kind of molecular labeling influencing duroc waistline character, SNP site correspond to international pig and refer to genome
T > C mutation on 10.2 edition 4 chromosomes at 3400805bp；
The SNP site of the molecular labeling of the influence duroc waistline character is that (being located at should shown in SEQ ID NO.1
The mutation of the 306th nucleic acid single base of sequence fragment, name are as follows: g.306T > C), wherein the M in sequence is T or C, leads to waistline character
Difference；
The molecular labeling of the influence duroc waistline character is in identification duroc waistline character and genetic breeding
Middle application；
A method of using the Duroc boar strain of the high lean meat percentage of above-mentioned molecular selection, comprise the following steps:
Detect the molecular labeling of above-mentioned influence duroc waistline character on No. 4 chromosomes of pig；The molecular labeling
SNP site is T or C, eliminates T and retains C；
The boar is preferably purebred American Duroc pig and its synthesis system (Guangdong Wen Shi boar Science and Technology Ltd.)；
It is a kind of for identifying the primer pair of the molecular labeling of above-mentioned influence duroc waistline character, comprising primer C-F and
C-R, nucleic acid sequence are as follows:
Primer C-F:5 '-GGGTGAGGGTGGGATTAG-3 '；
Primer C-R:5 '-TTGCGTTAGTGATTGTCCTG-3 '；
The primer pair influences to apply in duroc waistline character in identification；
The primer pair is applied in duroc molecular marker assisted selection breeding；
The primer pair is applied in the Duroc boar strain for cultivating high lean meat；
The method of the genetic improvement of one boar, comprises the following steps:
Determine the molecular labeling of the above-mentioned influence duroc waistline character of boar in nucleus herds of breeding pigs, and according to the upper of pig
The genotype of rheme point makes corresponding selection during boar is chosen seeds: the Systematic Breeding world pig of boar refers to genome
CC type individual on 10.2 edition 4 chromosomes at 3400805bp eliminates the TT type and CT type individual of the point；To be improved by generation
The frequency of the allele C in the site, thus the waistline of improved progeny pig；
The boar includes Duroc and its synthesis system；
The present invention has the following advantages and effects with respect to the prior art:
(1) present invention studies and determines the molecular labeling for influencing duroc waistline character, verifies it to waistline character
Influential effect, it is final to establish the molecular marker assisted selection breeding technique that quick improvement is carried out to waistline character, it significantly improves
Duroc and its synthetic figure breeding process adapt to the demand in live hog market, improve the price of pork pig, increase for enterprise
Add profit on sales, improves core competitiveness.
(2) present invention provides a kind of for identifying the primer pair for influencing the molecular labeling of duroc waistline character, passes through
The molecular labeling and primer pair can establish the molecular mark technology of efficiently and accurately, fast and accurately carry out to figure
Breeding process is accelerated in breeding.
(3) present invention provides a kind of method of figure breeding, energy by the advantage allele of the preferably molecular labeling
The genetic progress for enough accelerating duroc figure (waistline character), shortens the time of Duroc figure improvement, to effectively mention
The economic benefit of high kind of pig breeding, wherein by the molecular labeling, the present invention is individual at CC type by the whole breedings of TT type individual,
Then the whole waistline of Duroc group will increase 3cm, improve figure, improve meat yield.
Detailed description of the invention
Fig. 1 is that duroc is associated with (GWAS) analysis chart about the full-length genome of waistline character on No. 4 chromosomes；Its
In: the chromosome numbers of abscissa expression pig；Ordinate expression-logP value；
Fig. 2 is the waistline correlation analysis figure of different genotype pig.
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.
Embodiment 1
(1) experimental animal
It is Du Luo that experiment swinery group used in the present embodiment, which is the purebred U.S.A of Guangdong Wen Shi boar Science and Technology Ltd.,
Grams 627, be boar branch company core group, and group's pedigree record is detailed.
American Duroc pig boar in the sources group is chosen in this experiment altogether, and swinery is freely eaten, drinks water, entire feeding side
Formula, rearing conditions etc. are always consistent, are conventional method.
Duroc litter size is less, and big group mean is only 9~10, but grows fast, feed conversion rate height, trunk lean meat
Rate is high, and intramuscular fat content is higher, and resistance is strong.Domestic goods pig produces the more mature corss combination system of built standing.
Wherein the cross combination of Ternary Pig persistently keeps absolute superiority in the domestic and export markets in China.Since figure is
Unique evaluation criteria of living body market price, and its abdominal circumference and waistline etc. are huge for pig type score influence, so Duroc
The research that Pigs Kidney encloses character is particularly important.
(2) sample collection and phenotypic record
It collects the docking of above-mentioned boar and ear tissue is soaked in the ethyl alcohol that volume fraction is 75% that be placed in -20 DEG C of refrigerators standby
With.When pig is surveyed eventually, the waistline of pig is measured with tape, and has recorded 627 Duroc boar waistline data.
(3) pig full-length genome 50K SNP sentences type
The ear tissue or disconnected of each of 627 Duroc herd boars chosen from above-mentioned sources group individual acquisition
Tail tissue extracts complete genome DNA with standard phenol-chloroform method, quasi- through Nanodrop2000/2000C nucleic acid-protein detector
Really measure the concentration and OD ratio (OD260/280, OD260/230) of the DNA of every a sample.Through NanoDrop2000/2000C
DNA is diluted to 50ng/ μ L or so according to the concentration of detection by the DNA sample of nucleic acid-protein detector test qualification.Again by 6 μ L
The DNA sample to be measured extracted is mixed with 2 μ L Loading Buffer, is loaded to the Ago-Gel that mass fraction is 1%
In, electrophoresis 25min under 150V voltage observes and takes pictures under UV detector and gel imaging equipment, observes the complete of DNA
Whole property.
DNA sample Song Niuqin biotechnology (Shanghai) Co., Ltd., the basis on Illumina Beadstration platform
Company standard process carries out pig full-length genome 50K SNP chip (Illumina, the U.S.) genotype and determines.Utilize R language
Checkmarker carries out quality control to all sample 50K chip scanning typing datas in GenABEL packet, rejects detection individual
Rate is lower than 90%, family Mendel's error rate is higher than 0.1, minimum gene frequency less than 0.05 and Hardy-Weinberg equilibrium
Significance is higher than 10-6SNP, finally obtain the effective gene type data of 42642 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 waistline trait associations degree, and genomic level remarkable threshold removes for 0.05
With effective SNP site quantity, i.e. genome level of signifiance threshold value is 1.17 × 10-6, i.e., 0.05/42642 (effective SNP quantity)；
Chromosome level remarkable threshold is 1 divided by effective SNP site quantity, i.e. chromosome level of signifiance threshold value is 2.35 × 10-5, i.e., 1/
42642 (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. 4 chromosomes and significantly affecting in Duroc
The site that Pigs Kidney is enclosed, most strongly connected SNP are g.306T > C (P=7.70E-8)。
(5) association analysis of different genotype and waistline phenotype
According to table 1, the SNP site of molecular labeling g.306T > C is extremely significant related to waistline character, can be by right
The assisted Selection of this SNP site of duroc to improve group's waistline, and then accelerates breeding process.
It is also known according further to table 1, the average waistline of TT type ratio CC and CT type is low, illustrates that homozygote TT is to average waistline
It is worst.It is further learnt by Fig. 2, homozygote TT and CC and CT genotype significant difference, and CC and TT genotype is even more
Difference is extremely significant, this further illustrates that T allele is disadvantage allele.Waistline is the important indicator of pig appearance traits, waist
Girth illustrates that the figure of pig is sturdy.Therefore, more economic benefits can be brought by eliminating TT type and the pig of CT type, we are educating
The boar for needing to eliminate TT type and CT type during kind, retains the boar of CC, with the allele C by the generation raising site
Frequency.
The SNP site of 1 molecular labeling of table g.306T > correlation of C and waistline
2 target DNA sequence amplification of embodiment and sequencing
(1) design of primers
No. 4 chromosomes of pig 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.Design is drawn
The DNA sequence dna of object is as follows:
Primer C-F:5 '-GGGTGAGGGTGGGATTAG-3 '；
Primer C-R:5 '-TTGCGTTAGTGATTGTCCTG-3 '；
(2) PCR amplification
1 μ L of DNA profiling, 3.4 μ L, 2 × Tag PCR StanMixwith of distilled water are added in the reaction system of 10 μ L
Each 0.3 μ L of Loading Dye 5 μ L, primer C-F and C-R.PCR reaction condition are as follows: after 94 DEG C of initial denaturation 5min, 94 DEG C of denaturation
30s, 54.3 DEG C of annealing 30s, 72 DEG C of extension 45s, 35 circulations, last 72 DEG C of extensions 5min.
(3) determined dna sequence
Finally the product after PCR amplification is sequenced, sequencing is completed by Hua Da scientific & technical corporation, genetic fragment sequencing
It is required that logical for two-way survey.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.
The present invention is preliminary to carry out by detecting to the 306th bit base mutational site in SEQ ID NO.1 sequence
The application of association analysis between its genotype and the waistline character of pig provides one newly for the molecular marker assisted selection of pig
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.
SEQUENCE LISTING
<110>Guangdong Wen'S Foodstuffs Group Co., Ltd.；Agricultural University Of South China
<120>a kind of molecular labeling for influencing duroc waistline character and application
<130> 1
<160> 3
<170> PatentIn version 3.3
<210> 1
<211> 458
<212> DNA
<213> Artificial
<220>
<223>molecular labeling of duroc waistline character is influenced
<400> 1
gggtgagggt gggattagaa gtggtgccat gttggcgttc tctttgtaga aggcctagag 60
aggggatggc aggaagttat ggcttggaaa ttggatgagt gcggtggaaa caggtactaa 120
acggaaagca gaaagatctt ttatttactc ctttcttagt tgggcaacat ttctgggcca 180
cttgtcatgt tttcgtcagt gtactaagca tggagatagc cggatgaaca atgaagagtg 240
tatgcccttc aggagtttgc agatgaagga gcacacagac tcagacacca acgcctagag 300
cgtcgmgtga taagtgcttt ttgtggagat gcagagaaga gggctcctac cagagcttcc 360
tttgaacact gaattgaatc caggaggcgg tgcatacatt tgacatcttg tctcagaaat 420
gcaaaggtgg ctgaatacca ggacaatcac taacgcaa 458
<210> 2
<211> 18
<212> DNA
<213> Artificial
<220>
<223>primer C-F
<400> 2
gggtgagggt gggattag 18
<210> 3
<211> 20
<212> DNA
<213> Artificial
<220>
<223>primer C-R
<400> 3
ttgcgttagt gattgtcctg 20
Claims (9)
1. a kind of molecular labeling for influencing duroc waistline character encloses the application in character and pig genetics and breeding in identification Pigs Kidney,
It is characterized by: its SNP site corresponds to international pig with reference to the T at 3400805bp on 10.2 edition 4 chromosome of genome
> C mutation.
2. a kind of method of the boar strain using the high lean meat percentage of molecular selection for influencing duroc waistline character, special
Sign is to comprise the following steps:
Detect the molecular labeling for influencing duroc waistline character on No. 4 chromosomes of pig described in claim 1；Point
The SNP site of son label is T or C, eliminates T and retains C.
3. the boar according to claim 2 using the high lean meat percentage of molecular selection for influencing duroc waistline character
The method of strain, it is characterised in that:
The boar is purebred American Duroc pig and its synthesis system.
4. a kind of for identifying the primer pair for influencing the molecular labeling of duroc waistline character, it is characterised in that include primer C-
F and C-R, nucleic acid sequence are as follows:
Primer C-F:5 '-GGGTGAGGGTGGGATTAG-3 '；
Primer C-R:5 '-TTGCGTTAGTGATTGTCCTG-3 '；
The molecular labeling of the influence duroc waistline character is as shown in claim 1.
5. primer pair as claimed in claim 4 influences the application in duroc waistline character in identification.
6. application of the primer pair as claimed in claim 4 in duroc molecular marker assisted selection breeding.
7. application of the primer pair as claimed in claim 4 in the Duroc boar strain for cultivating high lean meat.
8. the method for the genetic improvement of a boar, characterized by comprising the steps of:
Determine the molecular labeling for influencing duroc waistline character in nucleus herds of breeding pigs described in the claim 1 of boar, and
Corresponding selection: the Systematic Breeding world pig ginseng of boar is made during boar is chosen seeds according to the genotype in the above-mentioned site of pig
The CC type individual on 10.2 edition 4 chromosome of genome at 3400805bp is examined, the TT type and CT type individual of the point are eliminated；With
The frequency that the allele C in the site is improved by generation, thus the waistline of improved progeny pig.
9. the method for the genetic improvement of pig according to claim 8, it is characterised in that:
The boar includes Duroc and its synthesis system.
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| CN110257529B (en) * | 2019-05-09 | 2022-07-05 | 华南农业大学 | SNP molecular marker related to lean meat percentage, eye muscle area and eye muscle thickness on pig No. 6 chromosome and application |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106906303A (en) * | 2017-04-27 | 2017-06-30 | 江西农业大学 | One SNP marker for influenceing quality character of pork and its application |
| CN107177681A (en) * | 2017-06-08 | 2017-09-19 | 华南农业大学 | Application of the pig SNP marker in daily gain in pigs behavior study and/or pig breeding |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106906303A (en) * | 2017-04-27 | 2017-06-30 | 江西农业大学 | One SNP marker for influenceing quality character of pork and its application |
| CN107177681A (en) * | 2017-06-08 | 2017-09-19 | 华南农业大学 | Application of the pig SNP marker in daily gain in pigs behavior study and/or pig breeding |
Non-Patent Citations (2)
| Title |
|---|
| Comparative analysis of QTLs influencing obesity and metabolic phenotypes in pigs and humans;PANT S D 等;《PLOS ONE》;20150908;1-22 |
| Complex relationship between obesity and the fat mass and obesity locus;YANG Q Y等;《INTERNATIONAL JOURNAL OF BIOLOGICAL SCIENCES》;20170515;第13卷;615-629 |
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