CN104164430A - Pork quality and meat yield related molecular markers and composite applications thereof - Google Patents
Pork quality and meat yield related molecular markers and composite applications thereof Download PDFInfo
- Publication number
- CN104164430A CN104164430A CN201410415756.2A CN201410415756A CN104164430A CN 104164430 A CN104164430 A CN 104164430A CN 201410415756 A CN201410415756 A CN 201410415756A CN 104164430 A CN104164430 A CN 104164430A
- Authority
- CN
- China
- Prior art keywords
- sequence
- igfbp6
- sdhd
- trim55
- individuality
- 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.)
- Pending
Links
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a group of pork quality and meat yield related molecular markers and composite applications thereof. The group of molecular markers disclosed by the invention comprises the following molecular markers shown as (1)-(3): (1) a molecular marker containing an IGFBP6-C44T polymorphic site, wherein the IGFBP6-C44T polymorphic site is the 830th site from 5' tail end in the IGFBP6 sequence and has the variation of C/T base. The group of molecular markers disclosed by the invention has the advantages that IGFBP6-C44T, TRIM55-C213T and SDHD-G131T molecular marker combinations are simultaneously identified, so that the synchronous improvement of pork quality and meat yield is realized and the breeding efficiency of molecular marker-assisted selection is greatly improved.
Description
Technical field
The present invention relates to one group of molecule marker and applied in any combination thereof; Be particularly related to a kind of molecule marker relevant to Meat and meat yield and applied in any combination thereof.
Background technology
Pork is the main source of animal proteinum, and China is traditional pork consumption big country, and pork is as Chinese traditional meat product, closely bound up with the raising of people's lives quality.China is one of country of raising pigs in the world the earliest, and the history of raising pigs, for several thousand, through hard raising and the seed selection of ancestors, has formed the local variety that differ from one another.Place of china kind has advantages of many outstanding, and such as meat is good, litter size is high, good stress resistance and strong adaptability etc., but has two common shortcomings: lean ratio is low and weightening finish is slow.External lean meat species kind and strain of cultivating, although feed conversion rate is high, growth fast, lean meat output is many, meat is bad, the probability that white muscles (PSE meat) occurs is high.Along with Chinese society expanding economy, the raising of living standards of the people and the level of consumption, the variation that has brought people's diet formula and diet idea, consumes outside quantitative requirement meeting pork, and human consumer is also more and more higher to the requirement of meat quality.Therefore, in the time improving the lean ratio of pig, how to take into account meat quality and become the emphasis of current pig breeding work.
Utilize quantitative genetics and traditional breeding method means to carry out genetic improvement to pig and obtained considerable progress, but traditional breeding method relies on the mensuration of boar performance index, often several months consuming time, management and feeding cost are increased, a number of simultaneously measuring is subject to the restriction of human and material resources and financial resources, seriously restricts the genetic improvement progress of pig.Therefore, find one easy and simple to handle, and can implement the early stage standard of selecting and evaluating, extremely important for the genetic breeding work of pig.Molecular biological develop rapidly makes the breeding work of pig occur new dawn.Utilize genetics or molecular biology method to carry out strong supplementing to the weak point of traditional breeding method work, also make the breeding of pig can break through the bottleneck problem that traditional breeding method work faces, promoted widely the development of pig breeding work.At present, marker assisted selection, the genome area location that affects pig important quantity proterties, the major gene that utilizes Genome Scanning Approach and the research of candidate gene method to affect pig important economical trait have successfully been applied in the middle of pig breeding practice both domestic and external, have obtained huge economic benefit and social benefit.By the enforcement of molecular breeding technology, the early stage selection of pig and directed fine breeding are become a reality gradually.
But for the quantitative character of meat and this quasi-representative of meat yield, the gene dosage that participates in regulation and control is huge, of a great variety, acting in conjunction and the impact mechanism of intergenic interaction on meat and meat yield is complicated, therefore can not only instruct the character improvement breeding of pig according to the variation of individual gene.Although existing business-like pig SNP chip, can realize by whole-genome association screening and the qualification of full genome molecule mark, but the cost of chip detection is relatively high at present, be unfavorable for the large-scale detection application in actual breeding.
Summary of the invention
The object of this invention is to provide the molecule marker relevant to Meat and meat yield and applied in any combination thereof.
The invention provides one group of molecule marker, formed by the molecule marker shown in following (1)-(3):
(1) molecule marker that contains IGFBP6-C44T pleomorphism site in IGFBP6 sequence, IGFBP6-C44T pleomorphism site is IGFBP6 sequence from 5 ' end the 830th, this site has the variation of C/T base;
The GenBank accession number of described IGFBP6 sequence is NM_001100190.1, and upgrading day is on January 10th, 2014;
(2) molecule marker that contains TRIM55-C213T pleomorphism site in TRIM55 sequence, TRIM55-C213T pleomorphism site is TRIM55 sequence from 5 ' end the 1999th, this site has the variation of C/T base;
The GenBank accession number of described TRIM55 sequence is XM_005663049.1, and upgrading day is on September 26th, 2013;
(3) molecule marker that contains SDHD-G131T pleomorphism site in SDHD sequence, SDHD-G131T pleomorphism site is SDHD sequence from 5 ' end the 444th, this site has the variation of G/T base;
The GenBank accession number of described SDHD sequence is NM_001097516.1, and upgrading day is on March 2nd, 2014.
In above-mentioned molecule marker, the sequence of the molecule marker that contains IGFBP6-C44T pleomorphism site in described IGFBP6 sequence is as shown in SEQ ID No.7;
The sequence of the molecule marker that contains TRIM55-C213T pleomorphism site in described TRIM55 sequence is as shown in SEQ IDNo.8;
The sequence of the molecule marker that contains SDHD-G131T pleomorphism site in described SDHD sequence is as shown in SEQ ID No.9.
One group of primer also belongs to protection scope of the present invention, is made up of the primer pair shown in following (1)-(3):
(1) DNA molecular shown in SEQ ID No.1 and SEQ ID No.2;
(2) DNA molecular shown in SEQ ID No.3 and SEQ ID No.4;
(3) DNA molecular shown in SEQ ID No.5 and SEQ ID No.6;
DNA molecular shown in described SEQ ID No.1 and SEQ ID No.2 is the primer of the DNA molecular shown in amplification SEQ ID No.7;
DNA molecular shown in described SEQ ID No.3 and SEQ ID No.4 is the primer of the DNA molecular shown in amplification SEQ ID No.8;
DNA molecular shown in described SEQ ID No.5 and SEQ ID No.6 is the primer of the DNA molecular shown in amplification SEQ ID No.9.
IGFBP6-C44T and/or the genotypic test kit of TRIM55-C213T and/or SDHD-G131T place of qualification or assistant identification pig also belong to a protection scope of the present invention, comprise the product shown in following (1)-(3):
(1) detect the product of the DNA molecular that contains IGFBP6-C44T pleomorphism site in IGFBP6 sequence, IGFBP6-C44T pleomorphism site is IGFBP6 sequence from 5 ' end the 830th, and this site has the variation of C/T base;
The GenBank accession number of described IGFBP6 sequence is NM_001100190.1, and upgrading day is on January 10th, 2014;
(2) detect the product of the DNA molecular that contains TRIM55-C213T pleomorphism site in TRIM55 sequence, TRIM55-C213T pleomorphism site is TRIM55 sequence from 5 ' end the 1999th, and this site has the variation of C/T base;
The GenBank accession number of described TRIM55 sequence is XM_005663049.1, and upgrading day is on September 26th, 2013;
(3) detect the product of the DNA molecular that contains SDHD-G131T pleomorphism site in SDHD sequence, SDHD-G131T pleomorphism site is SDHD sequence from 5 ' end the 444th, and this site has the variation of G/T base;
The GenBank accession number of described SDHD sequence is NM_001097516.1, and upgrading day is on March 2nd, 2014.
In mentioned reagent box, described product is above-mentioned primer.
IGFBP6-C44T and/or the genotypic method of TRIM55-C213T and/or SDHD-G131T place of qualification or assistant identification pig also belong to a protection scope of the present invention, comprise the step shown in following (1) and/or (2) and/or (3):
(1) genotype at IGFBP6-C44T place of qualification pig: taking the genomic dna of pig as template, taking the DNA molecular shown in SEQ ID No.1 and SEQ ID No.2 as primer, carry out pcr amplification, obtain pcr amplification product; Cut pcr amplification product with restriction enzyme TaaI enzyme, obtain enzyme and cut product; It is that the individuality that length is respectively two bands of 212bp and 44bp is that genotype is the individuality of CC that enzyme is cut product, it is that length is that the individuality of the band of 256bp is that genotype is the individuality of TT that enzyme is cut product, and it is that the individuality that length is respectively three bands of 256bp, 212bp and 44bp is that genotype is the individuality of TC that enzyme is cut product;
(2) genotype at TRIM55-C213T place of qualification pig: taking the genomic dna of pig as template, taking the DNA molecular shown in SEQ IDNo.3 and SEQ ID No.4 as primer, carry out pcr amplification, obtain pcr amplification product; Cut pcr amplification product with restriction enzyme Bsh1236I enzyme, obtain enzyme and cut product; It is that the individuality that length is respectively two bands of 212bp and 537bp is that genotype is the individuality of CC that enzyme is cut product, it is that length is that the individuality of the band of 749bp is that genotype is the individuality of TT that enzyme is cut product, and it is that the individuality that length is respectively three bands of 749bp, 537bp and 212bp is that genotype is the individuality of TC that enzyme is cut product;
(3) genotype at SDHD-G131T place of qualification pig: taking the genomic dna of pig as template, taking the DNA molecular shown in SEQ ID No.5 and SEQ ID No.6 as primer, carry out pcr amplification, obtain pcr amplification product; Cut pcr amplification product with restriction enzyme MboI enzyme, obtain enzyme and cut product; It is that the individuality that length is respectively two bands of 130bp and 90bp is that genotype is the individuality of GG that enzyme is cut product, it is that length is that the individuality of the band of 220bp is that genotype is the individuality of TT that enzyme is cut product, and it is that the individuality that length is respectively three bands of 220bp, 130bp and 90bp is that genotype is the individuality of TG that enzyme is cut product;
Described IGFBP6-C44T is IGFBP6 sequence from 5 ' end the 830th, and this site has the variation of C/T base; The GenBank accession number of described IGFBP6 sequence is NM_001100190.1, and upgrading day is on January 10th, 2014;
Described TRIM55-C213T is TRIM55 sequence from 5 ' end the 1999th, and this site has the variation of C/T base; The GenBank accession number of described TRIM55 sequence is XM_005663049.1, and upgrading day is on September 26th, 2013;
Described SDHD-G131T is SDHD sequence from 5 ' end the 444th, and this site has the variation of G/T base; The GenBank accession number of described SDHD sequence is NM_001097516.1, and upgrading day is on March 2nd, 2014.
The variation that a C/T base is contained at the 44th bit base place from 5 ' end of pcr amplification product in described (1), this site is described IGFBP6-C44T site, when this site base is C, being limited property restriction endonuclease TaaI identification enzyme are cut to two fragments that length is 212bp, 44bp;
The variation that a C/T base is contained at the 213rd bit base place from 5 ' end of pcr amplification product in described (2), this site is described TRIM55-C213T site, when this site base is C, being limited property restriction endonuclease Bsh1236I identification enzyme are cut to two fragments that length is 212bp, 537bp;
The variation that a G/T base is contained at the 131st bit base place from 5 ' end of pcr amplification product in described (3), this site is described SDHD-G131T site, when this site base is G, being limited property restriction endonuclease MboI identification enzyme are cut to two fragments that length is 130bp, 90bp.
In aforesaid method, the pcr amplification product in described (1) is as shown in SEQ ID No.7, and described IGFBP6-C44T is in the DNA molecular shown in SEQ ID No.7 from 5 ' end the 44th;
Pcr amplification product in described (2) is as shown in SEQ ID No.8, and described RIM55-C213T is in the DNA molecular shown in SEQ ID No.8 from 5 ' end the 213rd;
Pcr amplification product in described (3) is as shown in SEQ ID No.9, and described SDHD-G131T is in the DNA molecular shown in SEQ ID No.9 from 5 ' end the 131st;
The reaction system of described PCR is as follows: genomic dna 1 μ L, and 10 × pcr amplification damping fluid, 2.5 μ L, the each 0.75 μ L of described primer that concentration is 10mM, concentration is 10mM dNTPs 0.5 μ L, concentration is the Taq archaeal dna polymerase 0.25 μ L of 5U/ μ L, ddH
2o supplies system to 25 μ L;
The program of PCR in described (1) is as follows:
1. 94 DEG C of denaturation 5min;
2. 94 DEG C of sex change 40s;
3. 64 DEG C of annealing 30s;
4. 72 DEG C are extended 20s
5. repeat 2.～4. step 35 circulation;
6. last 72 DEG C are extended 5min;
The program of PCR in described (2) is as follows:
1. 94 DEG C of denaturation 5min;
2. 94 DEG C of sex change 40s;
3. 64 DEG C of annealing 30s;
4. 72 DEG C are extended 30s
5. repeat 2.～4. step 28 circulation;
6. last 72 DEG C are extended 5min;
The program of PCR in described (3) is as follows:
1. 94 DEG C of denaturation 3min;
2. 94 DEG C of sex change 15s; 66 DEG C of annealing 15s; 72 DEG C are extended 10s; 3 circulations;
3. 94 DEG C of sex change 15s; 64 DEG C of annealing 15s; 72 DEG C are extended 10s; 3 circulations;
4. 94 DEG C of sex change 15s; 60 DEG C of annealing 15s; 72 DEG C are extended 10s; 28 circulations;
5. last 72 DEG C are extended 3min.
The method that seed selection has the pig of good character also belongs to a protection scope of the present invention, is to select IGFBP6-C44T, TRIM55-C213T and SDHD-G131T site to be the genotypic individuality of TT to carry out seed selection;
Described IGFBP6-C44T is IGFBP6 sequence from 5 ' end the 830th; The GenBank accession number of described IGFBP6 sequence is NM_001100190.1, and upgrading day is on January 10th, 2014;
Described TRIM55-C213T is TRIM55 sequence from 5 ' end the 1999th; The GenBank accession number of described TRIM55 sequence is XM_005663049.1, and upgrading day is on September 26th, 2013;
Described SDHD-G131T is SDHD sequence from 5 ' end the 444th; The GenBank accession number of described SDHD sequence is NM_001097516.1, and upgrading day is on March 2nd, 2014;
Described good character is specially good meat yield performance and/or good Meat Quality;
Described good meat yield performance is specially chest lumbar vertebrae fat thickness, the triadic mean thickness of backfat, leg stern ratio, shoulder fat thickness, average fat thickness, the average thickness of backfat and/or eye muscle area;
Described good Meat Quality is specially marble grain scoring.
In aforesaid method, the described method of selecting IGFBP6-C44T, TRIM55-C213T and SDHD-G131T to be the genotypic individuality of TT is above-mentioned arbitrary described method.
Above-mentioned arbitrary described molecule marker, above-mentioned primer, the application of above-mentioned arbitrary described test kit in IGFBP6-C44T and/or the genotypic product of TRIM55-C213T and/or SDHD-G131T place of characterization or assistant identification pig also belong to protection scope of the present invention;
Described IGFBP6-C44T is IGFBP6 sequence from 5 ' end the 830th, and this site has the variation of C/T base; The GenBank accession number of described IGFBP6 sequence is NM_001100190.1, and upgrading day is on January 10th, 2014;
Described TRIM55-C213T is TRIM55 sequence from 5 ' end the 1999th, and this site has the variation of C/T base; The GenBank accession number of described TRIM55 sequence is XM_005663049.1, and upgrading day is on September 26th, 2013;
Described SDHD-G131T is SDHD sequence from 5 ' end the 444th, and this site has the variation of G/T base; The GenBank accession number of described SDHD sequence is NM_001097516.1, and upgrading day is on March 2nd, 2014;
Or,
The application that above-mentioned arbitrary described molecule marker, above-mentioned primer, above-mentioned arbitrary described test kit have in the pig of good character in seed selection also belongs to protection scope of the present invention;
Described good character is specially good meat yield performance and/or good Meat Quality;
Described good meat yield performance is specially chest lumbar vertebrae fat thickness, the triadic mean thickness of backfat, leg stern ratio, shoulder fat thickness, average fat thickness, the average thickness of backfat and/or eye muscle area;
Described good Meat Quality is specially marble grain scoring.
Beneficial effect of the present invention is: by IGFBP6-C44T, TRIM55-C213T and the combination of SDHD-G131T molecule marker are identified simultaneously, realize the synchronous improvement of Meat and meat yield proterties, greatly improved the breeding efficiency of molecular marker assisted selection.
Brief description of the drawings
Fig. 1 is the electrophorogram of three kinds of genotype (TT, TC, CC) of the TaaI-RFLPs of pig IGFBP6.
Fig. 2 is the electrophorogram of three kinds of genotype (TT, TC, CC) of the Bsh1236I-RFLPs of pig TRIM55.
Fig. 3 is the electrophorogram of three kinds of genotype (TT, TG, GG) of the MboI-RFLPs of pig SDHD.
Embodiment
The experimental technique using in following embodiment if no special instructions, is ordinary method.
Material, reagent etc. used in following embodiment, if no special instructions, all can obtain from commercial channels.
Piglet is purchased from Tongcheng County, Hubei Province stud farm.
The renewal day of the sequence that the GenBank accession number in following embodiment is NM_001100190.1 is on January 10th, 2014; GenBank accession number is to be on September 26th, 2013 the renewal day of the sequence of XM_005663049.1; GenBank accession number is to be on March 2nd, 2014 the renewal day of the sequence of NM_001097516.1.
The qualification of embodiment 1, Meat and the combination of meat yield molecule marker
One, the collection of sample
Be born the same day piglet, and taked the ear edge of pig or tail point to organize 0.2g left and right with pig overbit pincers or operating scissors, put into the centrifuge tube of 1.5mL, preserved for-20 DEG C long-term.
Two, tissue is shredded with scissors, with phenol chloroform extraction method extraction genomic dna.
Three, design of primers
Respectively with reference to its exon4 sequence of IGFBP6 gene (GenBank accession number NM_001100190.1), TRIM55 gene (GenBank accession number XM_005663049.1) its 3 ' UTR sequence and its exon4 sequence of sdhd gene (GenBank accession number NM_001097516.1), the primer of IGFBP6-C44T, the TRIM55-C213T that design amplification contains IGFBP6, TRIM55 and the corresponding pleomorphism site of SDHD and the molecule marker of SDHD-G131T, as shown in table 1.
Table 1 molecule marker fragment amplification primer
Four, pcr amplification reaction
(1) amplification of IGFBP6 molecule marker fragment
PCR reaction system (being totally 25 μ L): pig genomic dna 1 μ L, 10 × pcr amplification damping fluid is (containing Mg
2+) 2.5 μ L, 10mM primer I 6-s 0.75 μ L, 10mM primer I 6-a 0.75 μ L, 10mM dNTPs0.5 μ L, 5U/ μ L Taq archaeal dna polymerase 0.25 μ L, ddH
2o supplies system to 25 μ L.
PCR reaction conditions:
1. 94 DEG C of denaturation 5min;
2. 94 DEG C of sex change 40s;
3. 64 DEG C of annealing 30s;
4. 72 DEG C are extended 20s
5. repeat 2.～4. step 35 circulation;
6. last 72 DEG C are extended 5min.
Amplification obtains the sequence that length is 256bp, as shown in SEQ ID No.7.
(2) amplification of TRIM55 molecule marker fragment
PCR reaction system, as step (one), only replaces with primer T5-s and T5-a.
PCR reaction conditions:
1. 94 DEG C of denaturation 5min;
2. 94 DEG C of sex change 40s;
3. 64 DEG C of annealing 30s;
4. 72 DEG C are extended 30s
5. repeat 2.～4. step 28 circulation;
6. last 72 DEG C are extended 5min.
Amplification obtains the sequence that length is 749bp, as shown in SEQ ID No.8.
(3) amplification of SDHD molecule marker fragment
PCR reaction system, as step (one), only replaces with primer SD-s and SD-a.
PCR reaction conditions:
1. 94 DEG C of denaturation 3min;
2. 94 DEG C of sex change 15s; 66 DEG C of annealing 15s; 72 DEG C are extended 10s; 3 circulations;
3. 94 DEG C of sex change 15s; 64 DEG C of annealing 15s; 72 DEG C are extended 10s; 3 circulations;
4. 94 DEG C of sex change 15s; 60 DEG C of annealing 15s; 72 DEG C are extended 10s; 28 circulations;
5. last 72 DEG C are extended 3min.
Amplification obtains the sequence that length is 220bp, as shown in SEQ ID No.9.
Five, enzyme is cut Classification Identification
(1) qualification of IGFBP6 molecule marker fragment
Cut the DNA molecular shown in SEQ ID No.7 with restriction enzyme TaaI enzyme, enzyme is cut to product and carry out agarose gel electrophoresis qualification.In the DNA molecular variation that a C/T base is contained at the 44th bit base place from 5 ' end shown in SEQ ID No.7 (this site be positioned at IGFBP6 sequence (accession number NM_001100190.1) from 5 ' end the 830th), by this site called after IGFBP6-C44T, when this site base is C, can be identified by TaaI, it is two fragments of 212bp+44bp that the DNA molecular enzyme shown in SEQID No.7 is cut to length.
Electrophoresis result as shown in Figure 1.
Fig. 1 shows, CC type individuality contains two bands of 212bp+44bp, and TT type individuality only contains the band of a 256bp, and TC type individuality contains tri-bands of 256bp+212bp+44bp.
(2) qualification of TRIM55 molecule marker fragment
Cut the DNA molecular shown in SEQ ID No.8 with restriction enzyme Bsh1236I enzyme, enzyme is cut to product and carry out agarose gel electrophoresis qualification.In the DNA molecular variation that a C/T base is contained at 213bp place from 5 ' end shown in SEQ ID No.8 (this site be positioned at TRIM55 sequence (accession number XM_005663049.1) from 5 ' end the 1999th), by this site called after TRIM55-C213T, when this site base is C, can be identified by Bsh1236I, the DNA molecular enzyme shown in SEQ ID No.8 is cut to two fragments of 212bp+537bp.
Electrophoresis result as shown in Figure 2.
Fig. 2 shows, CC type individuality contains two bands of 212bp+537bp, and TT type individuality only contains the band of a 749bp, and TC type individuality contains tri-bands of 749bp+537bp+212bp.
(3) qualification of SDHD molecule marker fragment
Cut the DNA molecular shown in SEQ ID No.9 with restriction enzyme MboI enzyme, enzyme is cut to product and carry out agarose gel electrophoresis qualification.In the DNA molecular variation that a G/T base is contained at 131bp place from 5 ' end shown in SEQ ID No.9 (this site be positioned at SDHD sequence (accession number NM_001097516.1) from 5 ' end the 444th), called after SDHD-G131T, when this base sequence is G, can be identified by MboI, the DNA molecular enzyme shown in SEQ ID No.9 is cut to two fragments into 130bp+90bp.
Electrophoresis result as shown in Figure 3.
Fig. 3 shows, GG type individuality contains two bands of 130bp+90bp, and TT type individuality only contains the band of a 220bp, and TG type individuality contains tri-bands of 220bp+130bp+90bp.
The authentication information of above-mentioned three molecule markers is as shown in table 2.
The enzyme of table 2 molecule marker is cut qualification
Six, the selection of preponderant genotype individuality
According to the least square analytical method principle of variance analysis, generalized linear model (Generalized Linear Model, the GLM) program in application SPSS statistical package, carries out association analysis to polymorphic mark and proterties, as shown in table 3.The result of the pleomorphism site IGFBP6-C44T of pig IGFBP6exon4 and the analysis of part producing Properties Correlation shows: the chest lumbar vertebrae thickness of backfat of TT genotype individuality is significantly lower than CC genotype individuality; The triadic mean thickness of backfat of TT genotype individuality is extremely significantly lower than TC and CC genotype individuality; The leg stern ratio utmost point of TT genotype individuality is significantly higher than TC and CC genotype individuality.TT genotype is the preponderant genotype of the chest lumbar vertebrae thickness of backfat, the triadic mean thickness of backfat and leg stern ratio.
The result that the pleomorphism site TRIM55-C213T of pig TRIM553 ' UTR and part producing Properties Correlation are analyzed is as shown in table 4, and table 4 shows, the shoulder fat thickness of TT and TC genotype individuality is significantly lower than CC genotype individuality; The average fat thickness of TT and TC genotype individuality is significantly lower than CC genotype individuality; The average thickness of backfat of TC genotype individuality is significantly lower than CC genotype individuality; The leg stern ratio of TT and TC genotype individuality is significantly higher than CC genotype individuality.TT and TC genotype are the preponderant genotype of shoulder fat thickness, average fat thickness and leg stern ratio, the preponderant genotype that TC genotype is the average thickness of backfat.
The result that the pleomorphism site SDHD-G131T of pig SDHD exon4 and part producing Properties Correlation are analyzed is as shown in table 5, and table 5 shows, the eye muscle area of GG genotype individuality is extremely significantly greater than GT and TT genotype individuality; The marble grain scoring of TT genotype individuality is significantly higher than GT genotype individuality.GG genotype is the preponderant genotype of eye muscle area, and TT genotype is the preponderant genotype of marble grain scoring.
In sum, for IGFBP6-C44T pleomorphism site, in actual breeding process, choose TT genotype individuality, for the improvement of meat yield performance; For TRIM55-C213T pleomorphism site, can choose the improvement as meat yield performance of TT and TC genotype, but consider hereditary stability, therefore in actual breeding process, choose the TT genotype individuality isozygotying; For SDHD-G131T pleomorphism site, pay close attention to the improvement of Meat Quality, therefore in actual breeding process, choose TT genotype individuality, for the improvement of Meat Quality.
Chest lumbar vertebrae fat thickness in table 3, table 4 and table 5, the triadic mean thickness of backfat, leg stern ratio, shoulder fat thickness, average fat thickness, the average thickness of backfat, eye muscle area belong to meat yield performance, and marble grain scoring belongs to Meat Quality.
Table 3 IGFBP6-C44T and Part Traits association analysis table
* represent P<0.05, * * represents P<0.01
Table 4 TRIM55-C213T and Part Traits association analysis table
* represent P<0.05
Table 5 SDHD-G131T and Part Traits association analysis table
* represent P<0.05, * * represents P<0.01.
Claims (10)
1. one group of molecule marker, is made up of the molecule marker shown in following (1)-(3):
(1) molecule marker that contains IGFBP6-C44T pleomorphism site in IGFBP6 sequence, IGFBP6-C44T pleomorphism site is IGFBP6 sequence from 5 ' end the 830th, this site has the variation of C/T base;
The GenBank accession number of described IGFBP6 sequence is NM_001100190.1, and upgrading day is on January 10th, 2014;
(2) molecule marker that contains TRIM55-C213T pleomorphism site in TRIM55 sequence, TRIM55-C213T pleomorphism site is TRIM55 sequence from 5 ' end the 1999th, this site has the variation of C/T base;
The GenBank accession number of described TRIM55 sequence is XM_005663049.1, and upgrading day is on September 26th, 2013;
(3) molecule marker that contains SDHD-G131T pleomorphism site in SDHD sequence, SDHD-G131T pleomorphism site is SDHD sequence from 5 ' end the 444th, this site has the variation of G/T base;
The GenBank accession number of described SDHD sequence is NM_001097516.1, and upgrading day is on March 2nd, 2014.
2. molecule marker according to claim 1, is characterized in that: the sequence of the molecule marker that contains IGFBP6-C44T pleomorphism site in described IGFBP6 sequence is as shown in SEQ ID No.7;
The sequence of the molecule marker that contains TRIM55-C213T pleomorphism site in described TRIM55 sequence is as shown in SEQ IDNo.8;
The sequence of the molecule marker that contains SDHD-G131T pleomorphism site in described SDHD sequence is as shown in SEQ ID No.9.
3. one group of primer, is made up of the primer pair shown in following (1)-(3):
(1) DNA molecular shown in SEQ ID No.1 and SEQ ID No.2;
(2) DNA molecular shown in SEQ ID No.3 and SEQ ID No.4;
(3) DNA molecular shown in SEQ ID No.5 and SEQ ID No.6.
4. IGFBP6-C44T and/or the genotypic test kit of TRIM55-C213T and/or SDHD-G131T place of qualification or assistant identification pig, comprises the product shown in following (1)-(3):
(1) detect the product of the DNA molecular that contains IGFBP6-C44T pleomorphism site in IGFBP6 sequence, IGFBP6-C44T pleomorphism site is IGFBP6 sequence from 5 ' end the 830th, and this site has the variation of C/T base;
The GenBank accession number of described IGFBP6 sequence is NM_001100190.1, and upgrading day is on January 10th, 2014;
(2) detect the product of the DNA molecular that contains TRIM55-C213T pleomorphism site in TRIM55 sequence, TRIM55-C213T pleomorphism site is TRIM55 sequence from 5 ' end the 1999th, and this site has the variation of C/T base;
The GenBank accession number of described TRIM55 sequence is XM_005663049.1, and upgrading day is on September 26th, 2013;
(3) detect the product of the DNA molecular that contains SDHD-G131T pleomorphism site in SDHD sequence, SDHD-G131T pleomorphism site is SDHD sequence from 5 ' end the 444th, and this site has the variation of G/T base;
The GenBank accession number of described SDHD sequence is NM_001097516.1, and upgrading day is on March 2nd, 2014.
5. test kit according to claim 4, is characterized in that: described product is primer claimed in claim 3.
6. IGFBP6-C44T and/or the genotypic method of TRIM55-C213T and/or SDHD-G131T place of qualification or assistant identification pig, comprises the step shown in following (1) and/or (2) and/or (3):
(1) genotype at IGFBP6-C44T place of qualification pig: taking the genomic dna of pig as template, taking the DNA molecular shown in SEQ ID No.1 and SEQ ID No.2 as primer, carry out pcr amplification, obtain pcr amplification product; Cut pcr amplification product with restriction enzyme TaaI enzyme, obtain enzyme and cut product; It is that the individuality that length is respectively two bands of 212bp and 44bp is that genotype is the individuality of CC that enzyme is cut product, it is that length is that the individuality of the band of 256bp is that genotype is the individuality of TT that enzyme is cut product, and it is that the individuality that length is respectively three bands of 256bp, 212bp and 44bp is that genotype is the individuality of TC that enzyme is cut product;
(2) genotype at TRIM55-C213T place of qualification pig: taking the genomic dna of pig as template, taking the DNA molecular shown in SEQ IDNo.3 and SEQ ID No.4 as primer, carry out pcr amplification, obtain pcr amplification product; Cut pcr amplification product with restriction enzyme Bsh1236I enzyme, obtain enzyme and cut product; It is that the individuality that length is respectively two bands of 212bp and 537bp is that genotype is the individuality of CC that enzyme is cut product, it is that length is that the individuality of the band of 749bp is that genotype is the individuality of TT that enzyme is cut product, and it is that the individuality that length is respectively three bands of 749bp, 537bp and 212bp is that genotype is the individuality of TC that enzyme is cut product;
(3) genotype at SDHD-G131T place of qualification pig: taking the genomic dna of pig as template, taking the DNA molecular shown in SEQ ID No.5 and SEQ ID No.6 as primer, carry out pcr amplification, obtain pcr amplification product; Cut pcr amplification product with restriction enzyme MboI enzyme, obtain enzyme and cut product; It is that the individuality that length is respectively two bands of 130bp and 90bp is that genotype is the individuality of GG that enzyme is cut product, it is that length is that the individuality of the band of 220bp is that genotype is the individuality of TT that enzyme is cut product, and it is that the individuality that length is respectively three bands of 220bp, 130bp and 90bp is that genotype is the individuality of TG that enzyme is cut product;
Described IGFBP6-C44T is IGFBP6 sequence from 5 ' end the 830th, and this site has the variation of C/T base; The GenBank accession number of described IGFBP6 sequence is NM_001100190.1, and upgrading day is on January 10th, 2014;
Described TRIM55-C213T is TRIM55 sequence from 5 ' end the 1999th, and this site has the variation of C/T base; The GenBank accession number of described TRIM55 sequence is XM_005663049.1, and upgrading day is on September 26th, 2013;
Described SDHD-G131T is SDHD sequence from 5 ' end the 444th, and this site has the variation of G/T base; The GenBank accession number of described SDHD sequence is NM_001097516.1, and upgrading day is on March 2nd, 2014.
7. method according to claim 6, is characterized in that: the pcr amplification product in described (1) is as shown in SEQID No.7, and described IGFBP6-C44T is in the DNA molecular shown in SEQ ID No.7 from 5 ' end the 44th;
Pcr amplification product in described (2) is as shown in SEQ ID No.8, and described TRIM55-C213T is in the DNA molecular shown in SEQ IDNo.8 from 5 ' end the 213rd;
Pcr amplification product in described (3) is as shown in SEQ ID No.9, and described SDHD-G131T is in the DNA molecular shown in SEQ ID No.9 from 5 ' end the 131st.
8. seed selection has a method for the pig of good character, is to select IGFBP6-C44T, TRIM55-C213T and SDHD-G131T site to be the genotypic individuality of TT to carry out seed selection;
Described IGFBP6-C44T is IGFBP6 sequence from 5 ' end the 830th; The GenBank accession number of described IGFBP6 sequence is NM_001100190.1, and upgrading day is on January 10th, 2014;
Described TRIM55-C213T is TRIM55 sequence from 5 ' end the 1999th; The GenBank accession number of described TRIM55 sequence is XM_005663049.1, and upgrading day is on September 26th, 2013;
Described SDHD-G131T is SDHD sequence from 5 ' end the 444th; The GenBank accession number of described SDHD sequence is NM_001097516.1, and upgrading day is on March 2nd, 2014;
Described good character is specially good meat yield performance and/or good Meat Quality.
9. method according to claim 8, is characterized in that: the described method of selecting IGFBP6-C44T, TRIM55-C213T and SDHD-G131T to be the genotypic individuality of TT is the method described in claim 6 or 7.
10. the application of the test kit described in the molecule marker described in claim 1 or 2, primer claimed in claim 3, claim 4 or 5 in IGFBP6-C44T and/or the genotypic product of TRIM55-C213T and/or SDHD-G131T place of characterization or assistant identification pig;
Described IGFBP6-C44T is IGFBP6 sequence from 5 ' end the 830th, and this site has the variation of C/T base; The GenBank accession number of described IGFBP6 sequence is NM_001100190.1, and upgrading day is on January 10th, 2014;
Described TRIM55-C213T is TRIM55 sequence from 5 ' end the 1999th, and this site has the variation of C/T base; The GenBank accession number of described TRIM55 sequence is XM_005663049.1, and upgrading day is on September 26th, 2013;
Described SDHD-G131T is SDHD sequence from 5 ' end the 444th, and this site has the variation of G/T base; The GenBank accession number of described SDHD sequence is NM_001097516.1, and upgrading day is on March 2nd, 2014;
Or,
Test kit described in molecule marker described in claim 1 or 2, primer claimed in claim 3, claim 4 or 5 has the application in the pig of good character in seed selection;
Described good character is specially good meat yield performance and/or good Meat Quality.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410415756.2A CN104164430A (en) | 2014-08-21 | 2014-08-21 | Pork quality and meat yield related molecular markers and composite applications thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410415756.2A CN104164430A (en) | 2014-08-21 | 2014-08-21 | Pork quality and meat yield related molecular markers and composite applications thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104164430A true CN104164430A (en) | 2014-11-26 |
Family
ID=51908411
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410415756.2A Pending CN104164430A (en) | 2014-08-21 | 2014-08-21 | Pork quality and meat yield related molecular markers and composite applications thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104164430A (en) |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105255870A (en) * | 2015-10-30 | 2016-01-20 | 中国农业大学 | SNP (Single Nucleotide Polymorphism) molecular marker related with back fat thickness trait of pig and detection method of SNP molecular marker |
| CN107805665A (en) * | 2017-12-12 | 2018-03-16 | 吉林省农业科学院 | The related molecular labeling of Red Steppe physical signs and its application |
| CN107815500A (en) * | 2017-12-12 | 2018-03-20 | 吉林省农业科学院 | The related molecular labeling of Red Steppe meat and its application in meat identification |
| CN110106261A (en) * | 2019-05-30 | 2019-08-09 | 浙江大学 | The combination of the SNP marker of Jiaxing Black Pig and raw meat product and identification method |
| CN110106260A (en) * | 2019-05-30 | 2019-08-09 | 浙江大学 | The combination of the SNP marker of Fengjing pig and raw meat product and identification method |
| CN114196760A (en) * | 2020-09-18 | 2022-03-18 | 中国农业科学院农业基因组研究所 | Method for identifying or assisting in identifying growth traits of pigs and related SNP (single nucleotide polymorphism) markers thereof |
| CN117363741A (en) * | 2023-10-13 | 2024-01-09 | 湖北省农业科学院畜牧兽医研究所 | Molecular marker related to pig carcass and meat quality traits in pig GPX3 gene 5' UTR and application thereof |
| CN117778589A (en) * | 2023-12-25 | 2024-03-29 | 中国农业科学院深圳农业基因组研究所(岭南现代农业科学与技术广东省实验室深圳分中心) | Pig economic character associated mutation site detection reagent, kit and detection method |
| CN117778589B (en) * | 2023-12-25 | 2024-10-29 | 中国农业科学院深圳农业基因组研究所(岭南现代农业科学与技术广东省实验室深圳分中心) | Pig economic character associated mutation site detection reagent, kit and detection method |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1737138A (en) * | 2004-08-20 | 2006-02-22 | 李奎 | Pig ouclar muscles area character-related protein and its coding gene and uses |
| CN101418298A (en) * | 2008-10-30 | 2009-04-29 | 华中农业大学 | MuRF2 gene fragment cloning and application related to hog carcass characters as molecular marker |
-
2014
- 2014-08-21 CN CN201410415756.2A patent/CN104164430A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1737138A (en) * | 2004-08-20 | 2006-02-22 | 李奎 | Pig ouclar muscles area character-related protein and its coding gene and uses |
| CN101418298A (en) * | 2008-10-30 | 2009-04-29 | 华中农业大学 | MuRF2 gene fragment cloning and application related to hog carcass characters as molecular marker |
Non-Patent Citations (2)
| Title |
|---|
| 万斯妮: "猪IGFBP6和CYR61基因片段的克隆、定位、多态及组织表达谱研究", 《中国优秀硕士学位论文全文数据库 农业科技辑》 * |
| 张定校: "猪骨骼肌卫星细胞的分离培养及TRIM55基因的染色体定位、SNP检测和表达谱分析", 《中国优秀硕士学位论文全文数据库 农业科技辑》 * |
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105255870A (en) * | 2015-10-30 | 2016-01-20 | 中国农业大学 | SNP (Single Nucleotide Polymorphism) molecular marker related with back fat thickness trait of pig and detection method of SNP molecular marker |
| CN105255870B (en) * | 2015-10-30 | 2018-04-03 | 中国农业大学 | A kind of SNP marker and its detection method related to fat thickness at back of pig character |
| CN107805665A (en) * | 2017-12-12 | 2018-03-16 | 吉林省农业科学院 | The related molecular labeling of Red Steppe physical signs and its application |
| CN107815500A (en) * | 2017-12-12 | 2018-03-20 | 吉林省农业科学院 | The related molecular labeling of Red Steppe meat and its application in meat identification |
| CN110106261B (en) * | 2019-05-30 | 2020-11-24 | 浙江大学 | SNP marker combination and identification method of Jiaxing black pig and raw meat product |
| CN110106260A (en) * | 2019-05-30 | 2019-08-09 | 浙江大学 | The combination of the SNP marker of Fengjing pig and raw meat product and identification method |
| CN110106261A (en) * | 2019-05-30 | 2019-08-09 | 浙江大学 | The combination of the SNP marker of Jiaxing Black Pig and raw meat product and identification method |
| CN110106260B (en) * | 2019-05-30 | 2020-12-08 | 浙江大学 | SNP marker combination and identification method of Fengjing pigs and raw meat products |
| CN114196760A (en) * | 2020-09-18 | 2022-03-18 | 中国农业科学院农业基因组研究所 | Method for identifying or assisting in identifying growth traits of pigs and related SNP (single nucleotide polymorphism) markers thereof |
| CN114196760B (en) * | 2020-09-18 | 2024-04-05 | 中国农业科学院农业基因组研究所 | Method for identifying or assisting in identifying pig growth traits and related SNP markers thereof |
| CN117363741A (en) * | 2023-10-13 | 2024-01-09 | 湖北省农业科学院畜牧兽医研究所 | Molecular marker related to pig carcass and meat quality traits in pig GPX3 gene 5' UTR and application thereof |
| CN117778589A (en) * | 2023-12-25 | 2024-03-29 | 中国农业科学院深圳农业基因组研究所(岭南现代农业科学与技术广东省实验室深圳分中心) | Pig economic character associated mutation site detection reagent, kit and detection method |
| CN117778589B (en) * | 2023-12-25 | 2024-10-29 | 中国农业科学院深圳农业基因组研究所(岭南现代农业科学与技术广东省实验室深圳分中心) | Pig economic character associated mutation site detection reagent, kit and detection method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104164430A (en) | Pork quality and meat yield related molecular markers and composite applications thereof | |
| Li et al. | Analysis of genetic variation in eggplant and related Solanum species using sequence-related amplified polymorphism markers | |
| CN106636393B (en) | SNP marker and its application with pumpkin peel color gene linkage | |
| CN102485892A (en) | Traceablility SNP marker in pig GIGYF2 gene and detection method thereof | |
| CN110144418A (en) | A kind of C. olelfera SSR molecular marker primer and labeling method and application | |
| CN110343767B (en) | Specific primer of microsatellite molecular marker of litopenaeus vannamei and application of specific primer in genetic diversity analysis | |
| CN114774570B (en) | Molecular marker closely linked with wheat stem rot resistance QTL and application | |
| CN110129456A (en) | A kind of anti-vibrios molecular labeling combination of prawn and its application in breeding | |
| CN107502665A (en) | Differentiate the method for locust tree kind based on Capillary Electrophoresis fluorescence SSR finger-prints | |
| CN101818195B (en) | Genetic marker by taking pig miR-27a precursor flanking sequence SNP as trait of litter size of pig and application | |
| CN109234423B (en) | Application of soybean molecular marker and polymorphism thereof in identification of soybean branch number traits | |
| CN102168136B (en) | Application of LHCGR gene of Chinese Holstein cow used as molecular marker | |
| CN110541041B (en) | SNP marker related to Chinese domestic horse dwarf trait and application thereof | |
| CN116064904A (en) | Molecular marker closely linked with wheat stem rot resistance QTL Qfcr.sicau.2A and application | |
| CN103589715B (en) | One of pig AMY2 gene can be used for the SNP marker of tracing to the source and detection method thereof | |
| CN108220402A (en) | A kind of identification method of Chinese cabbage germplasm and kind genealogical relationship | |
| Sun et al. | Genetic variation in eight Chinese cattle breeds based on the analysis of microsatellite markers | |
| CN112695125B (en) | Katelia SSR molecular marker primer composition and application thereof | |
| CN104651523A (en) | Applications of single-stranded nucleotide sequence of INHA gene as animal superovulation molecule marker, and detection method | |
| CN103589716B (en) | One of pig SNCG gene can be used for the SNP marker of tracing to the source and detection method thereof | |
| CN103421771B (en) | Genetic marker for character of litter size of pig utilizing WIF1 gene | |
| CN116769891A (en) | SNP (Single nucleotide polymorphism) marker for rapidly identifying genetic sex of Chinese soft-shelled turtles as well as primer and application thereof | |
| CN106566872B (en) | The analysis method in the pig SNP marker site based on sequencing genotyping technique | |
| CN103397027A (en) | SNP marker for identification of red/green color of pear pericarp based on high resolution melting and application thereof | |
| Yan et al. | Analysis of pituitary specific transcription factor-1 gene polymorphism in several indigenous Chinese cattle and crossbred cattle |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20141126 |