CN107151692A - A kind of gene diagnosis kit for identifying the pig live body thickness of backfat - Google Patents
A kind of gene diagnosis kit for identifying the pig live body thickness of backfat Download PDFInfo
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- CN107151692A CN107151692A CN201610118638.4A CN201610118638A CN107151692A CN 107151692 A CN107151692 A CN 107151692A CN 201610118638 A CN201610118638 A CN 201610118638A CN 107151692 A CN107151692 A CN 107151692A
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- 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
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Abstract
The invention discloses a kind of gene diagnosis kit for identifying the pig live body thickness of backfat.It is following (1) or (2) or (3) or (4) that pig live body thickness of backfat method is identified using the gene diagnosis kit of the present invention：(1) it is to detect that the 460th deoxyribonucleotide of MEG3 genes of pig individual is T or A or T and A；(2) it is to detect that the 1137th deoxyribonucleotide of MEG3 gene Second Exons of pig individual is A or G or A and G；(3) it is to detect that the 1158th deoxyribonucleotide of MEG3 gene Second Exons of pig individual is A or G or A and G；(4) it is to detect that the 51st deoxyribonucleotide of MEG3 gene First Introns of pig individual is T or C or T and C.It is experimentally confirmed：The method of the present invention is consistent up to the practical measurement result of the 100kg body weight live body thickness of backfats with pig, to selecting excellent pig kind significant.
Description
Technical field
The invention belongs to genetic engineering field, and in particular to a kind of gene diagnosis kit of the identification pig live body thickness of backfat,
More particularly to a kind of method that the 100kg body weight live body thickness of backfats are reached based on MEG3 identified for genes or auxiliary identification pig.
Background technology
Pig is that pig industry occupies critical role in the world especially China's national economy first of six domestic animals.Pork is many
Proportion in meat products is always held at 1/3rd or so, is people than one of preferable non-staple foodstuff.The life of people
Living constantly to improve, traditional cultural technique can not meet demand of the people to pork.With GENERALIZATION OF MODERN BREEDING TECHNIQUE
Development, the production performance of pig significantly improves, and the healthy aquaculture of pig and fattens and just seems increasingly important.
In recent years, China is improved under lean meat percentage and growth rate, but the quality of pork by introducing high lean meat percentage pig kind
Drop and local flavor unification have caused the concern of consumer.And the quality of pork, local flavor are also dependent on the life of Animal muscles
Long and fat deposition.In actual production, according to national standard, back-fat thickness directly reacts the height of pork fat content
Low, for the swinery being corrected under same weight standard, back-fat thickness numerical value is bigger, then lean meat percentage is lower, bone
Flesh content is fewer.Therefore the judge index for weighing pig lean meat percentage can be used as with the thickness of backfat.Therefore, how effectively to carry
It is preceding to judge that pig back fat is the target that each pig farm operator and breeding scholar pursue.
Nearly ten years, the breeding for developing into pig of molecular biotechnology provides a kind of new something lost made a variation based on DNA
Pass mark.The particularly appearance of Molecular Marker Assisted Selection Technology, can to significantly improve that these quantitative characters of pig provide
Energy.
The content of the invention
First purpose of the present invention is to provide a kind of identification or auxiliary identification pig reaches 100kg body weight live body back fat thickness
Method.
What the present invention was provided identifies or aids in identify pig up to the method for 100kg body weight live body back fat thickness for as follows (1)
And/or (2) and/or (3) and/or (4)：
(1) it is to detect that the genotype of pig individual is TT genotype or AA genotype or TA genotype, according to
The genotype of the pig individual determines that the pig individual reaches 100kg body weight live body thickness of backfat sizes：The pig of AA genotype
Individual reaches 100kg up to pig individual of the 100kg body weight live bodies thickness of backfat more than TT genotype, the pig individual of TT genotype
Pig individual of the body weight live body thickness of backfat more than TA genotype；
The TT genotype is the homozygote that the 460th deoxyribonucleotide of MEG3 genes Second Exon is T；
The TA genotype be the 460th deoxyribonucleotide of MEG3 genes Second Exon be T and A it is miscellaneous
It is fit；
The AA genotype is the homozygote that the 460th deoxyribonucleotide of MEG3 genes Second Exon is A；
(2) it is to detect that the genotype of pig individual is AA genotype or GG genotype or AG genotype, according to
The genotype of the pig individual determines that the pig individual reaches 100kg body weight live body thickness of backfat sizes：The pig of AA genotype
Individual reaches 100kg up to pig individual of the 100kg body weight live bodies thickness of backfat more than GG genotype, the pig individual of GG genotype
Pig individual of the body weight live body thickness of backfat more than AG genotype；
The AA genotype is the homozygote that the 847th deoxyribonucleotide of MEG3 genes Second Exon is A；
The AG genotype be the 847th deoxyribonucleotide of MEG3 genes Second Exon be A and G it is miscellaneous
It is fit；
The GG genotype is the homozygote that the 847th deoxyribonucleotide of MEG3 genes Second Exon is G；
(3) it is to detect that the genotype of pig individual is AA genotype or GG genotype or AG genotype, according to
The genotype of the pig individual determines that the pig individual reaches 100kg body weight live body thickness of backfat sizes：The pig of AA genotype
Individual reaches 100kg up to pig individual of the 100kg body weight live bodies thickness of backfat more than GG genotype, the pig individual of GG genotype
Pig individual of the body weight live body thickness of backfat more than AG genotype；
The AA genotype is the homozygosis that the 1158th deoxyribonucleotide of MEG3 genes Second Exon is A
Body；
The AG genotype is that the 1158th deoxyribonucleotide of MEG3 genes Second Exon is A and G
Heterozygote；
The GG genotype is the homozygosis that the 1158th deoxyribonucleotide of MEG3 genes Second Exon is G
Body；
(4) it is to detect that the genotype of pig individual is TT genotype or CC genotype or TC genotype, according to
The genotype of the pig individual determines that the pig individual reaches 100kg body weight live body thickness of backfat sizes：The pig of TT genotype
Body reaches 100kg bodies up to pig individual of the 100kg body weight live bodies thickness of backfat more than CC genotype, the pig individual of CC genotype
Pig individual of the taskwork body thickness of backfat more than TC genotype；
The TT genotype is the homozygote that the 51st deoxyribonucleotide of MEG3 genes First Intron is T；
The TC genotype be the 51st deoxyribonucleotide of MEG3 genes First Intron be T and C it is miscellaneous
It is fit；
The CC genotype is the homozygote that the 51st deoxyribonucleotide of MEG3 genes First Intron is C；
The nucleotide sequence of the MEG3 genes Second Exon is as shown in sequence 1 in sequence table；
The nucleotide sequence of the MEG3 genes First Intron is as shown in sequence 2 in sequence table.
In the above method,
In (1), the genotype of the detection pig individual is AA genotype or GG genotype or AG bases
Because the method for type is following A) or B)：
A) direct Sequencing；
B the pcr amplification product containing the 460th deoxyribonucleotide of MEG3 genes Second Exon) is sequenced；
In (2), the genotype of the detection pig individual is AA genotype or GG genotype or AG bases
Because the method for type is following C) or D)：
C) direct Sequencing；
D the pcr amplification product containing the 1137th deoxyribonucleotide of MEG3 genes Second Exon) is sequenced；
In (3), the genotype of the detection pig individual is AA genotype or GG genotype or AG bases
Because the method for type is following E) or F)：
E) direct Sequencing；
F the pcr amplification product containing the 1158th deoxyribonucleotide of MEG3 genes Second Exon) is sequenced；
In (4), the genotype of the detection pig individual is TT genotype or CC genotype or TC bases
Because the method for type is following G) or H)：
G) direct Sequencing；
H the pcr amplification product containing the 51st deoxyribonucleotide of MEG3 genes First Intron) is sequenced；
The B), D) and F) in pcr amplification product used in primer for it is following 1) or 2)：
1) as the single stranded DNA shown in sequence 4 in the single strand dna shown in sequence in sequence table 3 and sequence table
The primer pair A of molecular composition；
2) primer pair being made up of the single strand dna shown in the single strand dna and sequence B shown in sequence A
C；
The sequence A has phase to be deleted sequence 3 or increasing or change one or several nucleotides with sequence 3
The nucleotides of congenerous；
The sequence B has phase to be deleted sequence 4 or increasing or change one or several nucleotides with sequence 4
The nucleotides of congenerous；
The H) in pcr amplification product used in primer for it is following 3) or 4)：
3) as the single stranded DNA shown in sequence 6 in the single strand dna shown in sequence in sequence table 5 and sequence table
The primer pair B of molecular composition；
4) primer pair being made up of the single strand dna shown in the single strand dna and sequence D shown in sequence C
D；
The sequence C has phase to be deleted sequence 5 or increasing or change one or several nucleotides with sequence 5
The nucleotides of congenerous；
The sequence D has phase to be deleted sequence 6 or increasing or change one or several nucleotides with sequence 6
The nucleotides of congenerous.
Second object of the present invention is to provide the 460th deoxidation of the MEG3 gene Second Exons of detection pig individual
The genotype of ribonucleotide and/or the 1137th deoxyribonucleotide and/or the 1158th deoxyribonucleotide
The thing of the genotype of 51st deoxyribonucleotide of the MEG3 gene First Introns of material or detection pig individual
The new application of matter.
The invention provides detection pig individual MEG3 gene Second Exons the 460th deoxyribonucleotide and
/ the 1137th deoxyribonucleotide and/or the 1158th deoxyribonucleotide genotype material or detection pig
The material of the genotype of 51st deoxyribonucleotide of the MEG3 gene First Introns of individual is in identification or auxiliary
Identification pig individual is helped to reach the application in the 100kg body weight live body thickness of backfats.
Present invention also offers the 460th deoxyribonucleotide of the MEG3 gene Second Exons of detection pig individual
And/or material or the detection of the genotype of the 1137th deoxyribonucleotide and/or the 1158th deoxyribonucleotide
The material of the genotype of 51st deoxyribonucleotide of the MEG3 gene First Introns of pig individual is preparing mirror
Fixed or auxiliary identification pig individual reaches the application in the product of the 100kg body weight live body thickness of backfats.
Present invention also offers the 460th deoxyribonucleotide of the MEG3 gene Second Exons of detection pig individual
And/or material or the detection of the genotype of the 1137th deoxyribonucleotide and/or the 1158th deoxyribonucleotide
The material of the genotype of 51st deoxyribonucleotide of the MEG3 gene First Introns of pig individual is in pig breeding
In application.
Present invention also offers the 460th deoxyribonucleotide of the MEG3 gene Second Exons of detection pig individual
And/or material or the detection of the genotype of the 1137th deoxyribonucleotide and/or the 1158th deoxyribonucleotide
The material of the genotype of 51st deoxyribonucleotide of the MEG3 gene First Introns of pig individual is preparing pig
Application in the product of breeding.
Present invention also offers the 460th deoxyribonucleotide of the MEG3 gene Second Exons of detection pig individual
And/or material or the detection of the genotype of the 1137th deoxyribonucleotide and/or the 1158th deoxyribonucleotide
The material of the genotype of 51st deoxyribonucleotide of the MEG3 gene First Introns of pig individual is thin in seed selection
Meat rate height and/or skeletal muscle content height and/or pig reach the application in the short boar of the 100kg body weight live body thickness of backfats.
Present invention also offers the 460th deoxyribonucleotide of the MEG3 gene Second Exons of detection pig individual
And/or material or the detection of the genotype of the 1137th deoxyribonucleotide and/or the 1158th deoxyribonucleotide
The material of the genotype of 51st deoxyribonucleotide of the MEG3 gene First Introns of pig individual is preparing choosing
Educate high lean meat percentage and/or skeletal muscle content height and/or pig is reached in the product of the short boar of the 100kg body weight live body thickness of backfats
Using.
Third object of the present invention is to provide the product that a kind of identification pig individual reaches the 100kg body weight live body thickness of backfats.
What the present invention was provided identifies pig individual up to the MEG3 that the product of the 100kg body weight live body thickness of backfats is detection pig individual
The 460th deoxyribonucleotide and/or the 1137th deoxyribonucleotide of gene Second Exon and/or
The of the MEG3 gene First Introns of the material of the genotype of 1158 deoxyribonucleotides or detection pig individual
The material of the genotype of 51 deoxyribonucleotides.
In above-mentioned application or product, the 460th deoxidation core of the MEG3 gene Second Exons of the detection pig individual
The thing of the genotype of ribotide and/or the 1137th deoxyribonucleotide and/or the 1158th deoxyribonucleotide
The material of the genotype of 51st deoxyribonucleotide of the MEG3 gene First Introns of matter or detection pig individual
For following X1)-X6)：
X1) as the single stranded DNA shown in sequence 4 in the single strand dna shown in sequence in sequence table 3 and sequence table
The primer pair A of molecular composition；
X2) as the single stranded DNA shown in sequence 6 in the single strand dna shown in sequence in sequence table 5 and sequence table
The primer pair B of molecular composition；
X3) the primer being made up of the single strand dna shown in the single strand dna and sequence B shown in sequence A
To C；
The sequence A has phase to be deleted sequence 3 or increasing or change one or several nucleotides with sequence 3
The nucleotides of congenerous；
The sequence B has phase to be deleted sequence 4 or increasing or change one or several nucleotides with sequence 4
The nucleotides of congenerous；
X4) the primer being made up of the single strand dna shown in the single strand dna and sequence D shown in sequence C
To D；
The sequence C has phase to be deleted sequence 5 or increasing or change one or several nucleotides with sequence 5
The nucleotides of congenerous；
The sequence D has phase to be deleted sequence 6 or increasing or change one or several nucleotides with sequence 6
The nucleotides of congenerous；
X5 X1) is contained) the primer pair A or X2) the primer pair B or X3) the primer pair C or X4)
The PCR reagent of the primer pair D；
X6 X1) is contained) the primer pair A or X2) the primer pair B or X3) the primer pair C or X4)
The primer pair D or X5) PCR reagent kit.
Fourth object of the present invention is to provide a kind of high seed selection lean meat percentage and/or skeletal muscle content height and/or pig up to 100
The method of the short boar of the kg body weight live body thickness of backfats.
The seed selection lean meat percentage that the present invention is provided is high and/or skeletal muscle content is high and/or pig reaches the 100kg body weight live body thickness of backfats
The method of short boar includes selection TC genotype and/or the pig of TA genotype and/or AG genotype carries out breeding；
The TA genotype be the 460th deoxyribonucleotide of MEG3 genes Second Exon be T and A it is miscellaneous
It is fit；
The TC genotype be the 51st deoxyribonucleotide of MEG3 genes First Intron be T and C it is miscellaneous
It is fit；
The AG genotype is the 1137th deoxyribonucleotide and/or the 1158th of MEG3 genes Second Exon
The heterozygote that position deoxyribonucleotide is A and G；
The nucleotide sequence of the MEG3 genes Second Exon is as shown in sequence 1 in sequence table；
The nucleotide sequence of the MEG3 genes First Intron is as shown in sequence 2 in sequence table.
Present invention finds T460A, A1137G and A1158G site of pig MEG3 gene Second Exons and
The T51C sites of MEG3 gene First Introns have a significant impact to pig up to the 100kg body weight live body thickness of backfats, and base
A kind of identification is provided in T51C, T460A, A1137G and A1158G site or auxiliary identification pig reaches 100kg bodies
The method of the taskwork body thickness of backfat.It is experimentally confirmed：The method of the present invention reaches the 100kg body weight live body thickness of backfats with pig
Practical measurement result is consistent, to selecting excellent pig kind significant.
Embodiment
Experimental method used in following embodiments is conventional method unless otherwise specified.
Material, reagent used etc., unless otherwise specified, are commercially obtained in following embodiments.
It is live body thickness of backfat when pig body weight reaches 100kg that pig in following embodiments, which reaches the 100kg body weight live bodies thickness of backfat,
Angle value.
Embodiment 1, a kind of method for aiding in identification pig to reach the 100kg body weight live body thickness of backfats
First, the screening of the SNP site of pig MEG3 genes
1st, the extraction of pig ear tissue sample DNA
Gather respectively after the blood sample of 32 Landraces from Hebei pig farm, sample collection, be stored in 75% wine
In essence, and in -20 DEG C of preservations.The genomic DNA of blood sample, specific steps are extracted using phenol-chloroform extraction method
It is as follows：
(1) preparation of ear tissue sample：The ear tissue sample that clip is of moderate size, the impurity on surface is fallen with normal saline flushing
And blood stains, it is put in 1.5ml centrifuge tubes, is shredded with eye scissors, and adds 500uL Tissue lysates (north thereto
Jing Baitaike Bioisystech Co., Ltd；Production code member：AU19011)；
(2) according to the size of the ear tissue sample of institute's clip, 15~25uL Proteinase K is added into centrifuge tube
(SIGMA-ALDRICH；Production code member：SRE0005)；
(3) it is put on shaking bath, 55 DEG C of digestion are stayed overnight, it is ensured that the digestion of ear tissue sample is complete, obtains digesting thoroughly
Ear tissue；
(4) by above-mentioned digestion, thoroughly ear tissue sample is placed in room temperature, adds isometric Tris- saturated phenols (Jiangsu
Bao Lai bio tech ltd), overturn reverse mixing 5min；
(5) 14000rpm/min, which is centrifuged, occurs lamination in 10min, centrifuge tube, upper strata is to contain nucleic acid
Aqueous phase, lower floor is the organic phase containing other impurities.
(6) aqueous phase of the upper strata containing nucleic acid is carefully drawn using micropipettor, a new 1.5ml centrifuge tube is positioned over
In (in order to avoid picking up the organic phase of lower floor, preferably with the blue electron gun head for removing tip), abandon lower floor's organic phase；
(7) repeat step (4)~(6)；
(8) isometric saturated phenol is added into the centrifuge tube containing aqueous phase：Chloroform=1：1, overturn repeatedly and mix 5min,
14000rpm/min centrifuges 10min, then draws supernatant liquid, is put in a new 1.5mL centrifuge tube；
(9) repeat step (8)；
(10) saturated phenol is added thereto：Chloroform：The volume ratio of isoamyl alcohol is 25：24：1, reverse mixing is overturned,
14000rpm/min centrifuges 10min；
(11) supernatant liquid is absorbed, the absolute ethyl alcohol (being put in precooling in -20 DEG C of refrigerators in advance) for adding 2 times of volumes sinks
Thread or cotton-shaped DNA precipitations, can now occur in centrifuge tube in shallow lake DNA；
(12) the thread or cotton-shaped DNA that chooses with yellow pipette tips carefully is precipitated, and is put in new 1.5mL EP
Pipe, adds 70% ethanol (- 20 DEG C of precoolings) 500uL thereto, and washing DNA precipitations gently overturn 30s, abandoned
70% ethanol；
(13) repeat step (12)；
(14) it will be put in containing the centrifuge tube that DNA is precipitated in room temperature environment, dry 20~30min；
(15) 60~80uL TE buffer solutions or distilled water dissolving DNA precipitation (not acutely vibration or centrifugation) is added,
It is stored in -20 DEG C of refrigerators.
2nd, the amplification and sequencing of purpose fragment
(1) PCR is expanded
Using step 1 obtain genomic DNA as template, be respectively adopted primer pair F1 and R1 and primer pair F2 and
R2 enters performing PCR amplification, obtains pcr amplification product.Primer pair F1 and R1 amplification obtain the of MEG3 genes
Two extrons (sequence 1), primer pair F2 and R2 expand the First Intron (sequence 2) for obtaining MEG3 genes.
Primer sequence is as follows：
F1：ATGCCTGACGCAGTAACAA (sequence 3)；
R1：CCGCTTCCTTCATCCCTC (sequence 4)；
F2：AGCGACGTATTTAAGGT (sequence 5)；
R2：TCTCCCACAGCACTCC (sequence 6).
PCR amplification system：10 × LA PCR Buffer 2ul, 10mM dNTP Mix 1.6ul, upstream and downstream primer (10
Pmol/L) each 1ul, LA Taq archaeal dna polymerase (5U/ul), genomic DNA 1ul, ddH2O 13.2ul.
PCR response procedures：94 DEG C of pre-degenerations, 5min；94 DEG C of denaturation, 30s, 58 DEG C of annealing, 30s, 72 DEG C are prolonged
Stretch, 30s, 72 DEG C of extensions, 10min.
(2) recovery purifying of PCR primer
Utilize agarose gel purification QIAquick Gel Extraction Kit (the Tyke Bioisystech Co., Ltd of Beijing hundred), recovery purifying PCR
Product, concrete operation step is according to the incidental specification of the kit.
(3) coupled reaction
The pcr amplification product that above-mentioned purifying is reclaimed is attached with pMD18-T carriers.Coupled reaction system is
5μL：PCR recovery product 2ul, pMD18-T carrier 0.5ul, Solution I 2.5ul, 4 DEG C of connections are stayed overnight, obtained
Connection product.
(4) convert
Connection product is converted into bacillus coli DH 5 alpha competent cell.Concrete operation step：From -80 DEG C of ultra low temperature freezers
1 DH5 α competent cell (100 μ L) is inside taken, is put in rapidly on ice, melts it；By connection product (volume
5uL) add in competent cell, gently blown and beaten repeatedly with pipettor, mix it, be positioned on ice, stand 30
min；It is positioned in 42 DEG C of water-baths, thermal shock 90s, immediately after 2~3min of ice bath；Add 600 μ L's thereto
LB liquid medium (is free of ampicillin)；It is positioned in 37 DEG C of constant-temperature tables, with turning for 170~200r/min
Speed culture 1h, makes bacteria resuscitation.
(5) positive clone identification
Bacterium solutions of 50~100 μ L by recovery is drawn with pipettor, the agar containing ampicillin is spread evenly across and puts down
On plate, after bacterium solution is absorbed completely, 30min in 37 DEG C of constant incubators is put in, bacterium solution is absorbed completely, so
Agar plate is inverted afterwards and is put in constant incubator, 37 DEG C of overnight incubations.
According to the colony growth situation on agar plate, bacterium is chosen in progress.1mL liquid is added into 1.5mL EP pipes
LB culture mediums, while 2 μ L ampicillin is added into each EP pipes, then with 10 μ L pipette tips picking agar
White colony (selected shape is complete, in the bacterium colony of circular point shape) totally 20 on flat board, is put in fills 1mL respectively
In the 1.5mL EP pipes of fluid nutrient medium, it is put in 37 DEG C of shaking tables, 3~4h of culture is expanded with 220r/min rotating speed,
When there is turbid phenomenon or white thread precipitation in EP pipes, using using primer pair F1 and R1 and primer pair F2
Bacterium solution PCR identifications are carried out with R2.
PCR reaction systems are as follows：10 × LA Buffer 1ul, dNTP Mix (2.5mM) 0.8ul, upstream and downstream primer
(pmol/L) each 0.5ul, LA polymerase 0.1ul, bacterium solution 0.5ul, ddH2O 6.6ul, total system is 10ul.
After PCR reactions terminate, draw 1 μ L PCR primers and carry out 1.5% agarose gel electrophoresis detection, agarose coagulates
Glue imager is taken pictures, and primarily determines that positive colony.
(6) sequence verification
The bacterium solution for being initially identified as positive colony by PCR amplifications is delivered into Shanghai Invitrogen Co., Ltd to be surveyed
Sequence.
(7) acquisition of SNP site (T51C, T460A, A1137G and A1158G)
According to above-mentioned multiple sequencing results, it is compared using DNAman softwares, obtaining 4 difference sites is
T460A (the 460th nucleotides of sequence 1), A1137G (the 1137th nucleotides of sequence 1), A1158G
(the 1158th nucleotides of sequence 1) and T51C (the 51st nucleotides of sequence 2).
Base in the T460A sites of pig MEG3 genes Second Exon (sequence 1) is T individual, this
Body is homozygous individual, and the individual genotype is named as into homozygosis TT genotype, and pig MEG3 genes second are outer aobvious
The base in the T460A sites of sub (sequence 1) is A individual, and the individual is homozygous individual, by the individual
Genotype is named as homozygosis AA genotype, the T460A sites of pig MEG3 genes Second Exon (sequence 1)
Base is T and A individual, and the individual is heterozygous individual, and the individual genotype is named as into heterozygosis TA genes
Type.
Base in the A1137G sites of pig MEG3 genes Second Exon (sequence 1) is A individual, should
Individual is homozygous individual, and the individual genotype is named as outside homozygosis AA genotype, pig MEG3 genes second
The base in the A1137G sites of aobvious son (sequence 1) is G individual, and the individual is homozygous individual, by this
The genotype of body is named as homozygosis GG genotype, the A1137G of pig MEG3 genes Second Exon (sequence 1)
The base in site is A and G individual, and the individual is heterozygous individual, and the individual genotype is named as into heterozygosis
AG genotype.
Base in the A1158G sites of pig MEG3 genes Second Exon (sequence 1) is A individual, should
Individual is homozygous individual, and the individual genotype is named as outside homozygosis AA genotype, pig MEG3 genes second
The base in the A1158G sites of aobvious son (sequence 1) is G individual, and the individual is homozygous individual, by this
The genotype of body is named as homozygosis GG genotype, the A1158G of pig MEG3 genes Second Exon (sequence 1)
The base in site is A and G individual, and the individual is heterozygous individual, and the individual genotype is named as into heterozygosis
AG genotype.
Base in the T51C sites of pig MEG3 genes First Intron (sequence 2) is T individual, this
Body is homozygous individual, the individual genotype is named as into homozygosis TT genotype, pig MEG3 genes first are included
The base in the T51C sites of sub (sequence 2) is C individual, and the individual is homozygous individual, by the individual
Genotype is named as homozygosis CC genotype, the alkali in the T51C sites of pig MEG3 genes First Intron (sequence 2)
Base is T and C individual, and the individual is heterozygous individual, and the individual genotype is named as into heterozygosis TC genotype.
2nd, T51C, T460A, A1137G, A1158G of pig MEG3 genes and pig reach 100kg body weight live bodies
The correlation analysis of the thickness of backfat
(1) the T460A sites of pig MEG3 genes Second Exon reach the phase of the 100kg body weight live body thickness of backfats with pig
The analysis of closing property
To determine whether the T460A sites of pig MEG3 gene Second Exons reach the 100kg body weight live bodies thickness of backfat with pig
Correlation, 294 Large Whites using Hebei pig farm determine each pig as experiment material according to the method in above-mentioned steps one
The genotype of individual is TT genotype, AA genotype or TA genotype, and determines that pig reaches according to the genotype of pig
The 100kg body weight live body thickness of backfats：The pig individual of AA genotype is more than TT genotype up to the 100kg body weight live bodies thickness of backfat
Pig individual, TT genotype pig individual up to the 100kg body weight live bodies thickness of backfat more than TA genotype pig individual.
1st, genotype
The genotype call results of 294 pig individuals show：The genotype of 288 pigs is TT genotype, 2 pigs
Genotype is AA genotype, and the genotype of 4 pigs is TA genotype.Pig MEG3 genes are in detection swinery
Genotype frequency and gene frequency result are as shown in table 1：As can be seen from Table 1：Genotype frequency homozygous TT
Rate is apparently higher than AA is homozygous and TA heterozygous, and T allele is protogene.
The genotype frequency and gene frequency of table 1, pig MEG3 genes in detection swinery
2nd, pig genotype and pig reach the correlation analysis of the 100kg body weight live body thickness of backfats
Statistical analysis is carried out up to the 100kg body weight live bodies thickness of backfat to genotype and pig using the softwares of SPSS 20.0, and done
Multiple range test between sample.
As a result it is as shown in table 2：SNP (T460A sites) sites have significantly to pig up to the 100kg body weight live body thickness of backfats
Influence, the pig of AA genotype is individual more than the pig of TT genotype up to the 100kg body weight live bodies thickness of backfat, TT genotype
Pig up to the 100kg body weight live bodies thickness of backfat more than TA genotype pig individual, and AA genotype pig reach 100kg
The body weight live body thickness of backfat is significantly more than TA genotype 7.17mm or so.
Table 2, pig MEG3 genes single nucleotide polymorphism are with reaching 100kg body weight live body thickness of backfat correlation analyses
Remarks:With different lowercase letter indication differences significantly (P in table<0.05), same letter represents difference not significantly (P>0.05), numerical value least square mean value ± standard error.
In summary, it can be determined by the nucleotides in the T460A sites for determining pig MEG3 gene Second Exons
Pig individual is TT genotype or TA genotype or AA genotype, so as to aid in identification pig to be lived up to 100kg body weight
The body thickness of backfat：The pig of AA genotype is individual more than the pig of TT genotype up to the 100kg body weight live bodies thickness of backfat, TT bases
Because the pig of type is individual more than the pig of TA genotype up to the 100kg body weight live bodies thickness of backfat；
TT genotype be pig MEG3 genes Second Exon the 460th nucleotides be T homozygote；
AA genotype be pig MEG3 genes Second Exon the 460th nucleotides be A homozygote；
TA genotype is that the nucleotides of pig MEG3 genes Second Exon the 460th is T and A heterozygote；
The nucleotide sequence of MEG3 gene Second Exons is as shown in sequence 1 in sequence table.
(2) the A1137G sites of pig MEG3 genes Second Exon reach the 100kg body weight live body thickness of backfats with pig
Correlation analysis
It is to determine that the A1137G sites of pig MEG3 gene Second Exons reach the 100kg body weight live body thickness of backfats with pig
No correlation, 294 Large Whites using Hebei pig farm determine each as experiment material according to the method in above-mentioned steps one
The genotype of pig individual is AA genotype, GG genotype or AG genotype, and determined according to the genotype of pig
Pig reaches the 100kg body weight live body thickness of backfats：The pig individual of AA genotype is more than GG up to the 100kg body weight live bodies thickness of backfat
The pig individual of genotype, the pig individual of GG genotype is individual more than the pig of AG genotype up to the 100kg body weight live bodies thickness of backfat
Body.
1st, genotype
The genotype call results of 294 pig individuals show：The genotype of 26 pigs is AA genotype, 164 pigs
Genotype be GG genotype, the genotype of 104 pigs is AG genotype.Pig MEG3 genes are in detection swinery
In genotype frequency and gene frequency result it is as shown in table 3：As can be seen from Table 3：GG is homozygous and AG
The genotype frequency of heterozygous apparently higher than AA it is homozygous and, G allele be protogene.
The genotype frequency and gene frequency of table 3, pig MEG3 genes in detection swinery
2nd, pig genotype and pig reach the correlation analysis of the 100kg body weight live body thickness of backfats
Statistical analysis is carried out up to the 100kg body weight live bodies thickness of backfat to genotype and pig using the softwares of SPSS 20.0, and done
Multiple range test between sample.
As a result it is as shown in table 4：SNP (A1137G sites) sites have significantly to pig up to the 100kg body weight live body thickness of backfats
Influence, the pig of AA genotype is individual more than the pig of GG genotype up to the 100kg body weight live bodies thickness of backfat, TT genotype
Pig up to the 100kg body weight live bodies thickness of backfat more than AG genotype pig individual, and AA genotype pig reach 100kg
The body weight live body thickness of backfat is significantly more than AG genotype 1.23mm or so.
Table 4, pig MEG3 genes single nucleotide polymorphism are with reaching 100kg body weight live body thickness of backfat correlation analyses
Remarks:With different lowercase letter indication differences significantly (P in table<0.05), same letter represents difference not significantly (P>0.05), numerical value least square mean value ± standard error.
In summary, can by determine pig MEG3 gene Second Exons A1137G sites nucleotides come really
It is AA genotype or GG genotype or AG genotype to determine pig individual, so as to aid in identification pig to reach 100kg bodies
The taskwork body thickness of backfat：The pig of AA genotype is individual more than the pig of GG genotype up to the 100kg body weight live bodies thickness of backfat,
The pig of TT genotype is individual more than the pig of AG genotype up to the 100kg body weight live bodies thickness of backfat；
AA genotype be pig MEG3 genes Second Exon the 1137th nucleotides be A homozygote；
GG genotype be pig MEG3 genes Second Exon the 1137th nucleotides be G homozygote；
AG genotype is that the nucleotides of pig MEG3 genes Second Exon the 1137th is A and G heterozygote；
The nucleotide sequence of MEG3 gene Second Exons is as shown in sequence 1 in sequence table.
(3) the A1158G sites of pig MEG3 genes Second Exon reach the 100kg body weight live body thickness of backfats with pig
Correlation analysis
It is to determine that the A1158G sites of pig MEG3 gene Second Exons reach the 100kg body weight live body thickness of backfats with pig
No correlation, 295 Large Whites using Hebei pig farm determine each as experiment material according to the method in above-mentioned steps one
The genotype of pig individual is AA genotype, GG genotype or AG genotype, and determined according to the genotype of pig
Pig reaches the 100kg body weight live body thickness of backfats：The pig individual of AA genotype is more than GG up to the 100kg body weight live bodies thickness of backfat
The pig individual of genotype, the pig individual of GG genotype is individual more than the pig of AG genotype up to the 100kg body weight live bodies thickness of backfat
Body.
1st, genotype
The genotype call results of 295 pig individuals show：The genotype of 25 pigs is AA genotype, 164 pigs
Genotype be GG genotype, the genotype of 106 pigs is AG genotype.Pig MEG3 genes are in detection swinery
In genotype frequency and gene frequency result it is as shown in table 1：As can be seen from Table 5：GG is homozygous and AG
The genotype frequency of heterozygous is homozygous apparently higher than AA, and G allele is protogene.
The genotype frequency and gene frequency of table 5, pig MEG3 genes in detection swinery
2nd, pig genotype and pig reach the correlation analysis of the 100kg body weight live body thickness of backfats
Statistical analysis is carried out up to the 100kg body weight live bodies thickness of backfat to genotype and pig using the softwares of SPSS 20.0, and done
Multiple range test between sample.
As a result it is as shown in table 6：SNP (A1158G sites) sites have significantly to pig up to the 100kg body weight live body thickness of backfats
Influence, the pig of AA genotype is individual more than the pig of GG genotype up to the 100kg body weight live bodies thickness of backfat, GG genotype
Pig up to the 100kg body weight live bodies thickness of backfat more than AG genotype pig individual, and AA genotype pig reach 100kg
The body weight live body thickness of backfat is significantly more than AG genotype 1.25mm or so.
Table 6, pig MEG3 genes single nucleotide polymorphism are with reaching 100kg body weight live body thickness of backfat correlation analyses
Remarks:With different lowercase letter indication differences significantly (P in table<0.05), same letter represents difference not significantly (P>0.05), numerical value least square mean value ± standard error.
In summary, can by determine pig MEG3 gene Second Exons A1158G sites nucleotides come really
It is AA genotype or GG genotype or AG genotype to determine pig individual, so as to aid in identification pig to reach 100kg bodies
The taskwork body thickness of backfat：The pig of AA genotype is individual more than the pig of GG genotype up to the 100kg body weight live bodies thickness of backfat,
The pig of GG genotype is individual more than the pig of AG genotype up to the 100kg body weight live bodies thickness of backfat；
AA genotype be pig MEG3 genes Second Exon the 1158th nucleotides be A homozygote；
GG genotype be pig MEG3 genes Second Exon the 1158th nucleotides be G homozygote；
AG genotype is that the nucleotides of pig MEG3 genes Second Exon the 1158th is A and G heterozygote；
The nucleotide sequence of MEG3 gene Second Exons is as shown in sequence 1 in sequence table.
(4) the T51C sites of pig MEG3 genes First Intron reach the phase of the 100kg body weight live body thickness of backfats with pig
The analysis of closing property
To determine whether the T51C sites of pig MEG3 gene First Introns reach the 100kg body weight live bodies thickness of backfat with pig
Correlation, 295 Large Whites using Hebei pig farm determine each pig as experiment material according to the method in above-mentioned steps one
The genotype of individual is TT genotype, CC genotype or TC genotype, and determines that pig reaches according to the genotype of pig
The 100kg body weight live body thickness of backfats：The pig individual of TT genotype is more than CC genotype up to the 100kg body weight live bodies thickness of backfat
Pig individual, CC genotype pig individual up to the 100kg body weight live bodies thickness of backfat more than TC genotype pig individual.
1st, genotype
The genotype call results of 295 pig individuals show：The genotype of 26 pigs is TT genotype, 163 pigs
Genotype is CC genotype, and the genotype of 106 pigs is TC genotype.Pig MEG3 genes are in detection swinery
Genotype frequency and gene frequency result are as shown in table 1：As can be seen from Table 7：CC is homozygous and CT heterozygosis
The genotype frequency of type is homozygous apparently higher than TT, and C allele is protogene.
The genotype frequency and gene frequency of table 7, pig MEG3 genes in detection swinery
2nd, pig genotype and pig reach the correlation analysis of the 100kg body weight live body thickness of backfats
Statistical analysis is carried out up to the 100kg body weight live bodies thickness of backfat to genotype and pig using the softwares of SPSS 20.0, and done
Multiple range test between sample.
As a result it is as shown in table 8：SNP (T51C) sites have a significant impact to pig up to the 100kg body weight live body thickness of backfats,
The pig of TT genotype reaches up to pig individual of the 100kg body weight live bodies thickness of backfat more than CC genotype, the pig of CC genotype
The 100kg body weight live bodies thickness of backfat is individual more than the pig of TC genotype, and the pig of TT genotype reaches 100kg body weight live bodies
The thickness of backfat is significantly more than TC genotype 1.46mm or so.
Table 8, pig MEG3 genes single nucleotide polymorphism are with reaching 100kg body weight live body thickness of backfat correlation analyses
Remarks:With different lowercase letter indication differences significantly (P in table<0.05), same letter represents difference not significantly (P>0.05), numerical value least square mean value ± standard error.
In summary, it can be determined by the nucleotides in the T51C sites for determining pig MEG3 gene First Introns
Pig individual is TT genotype or CC genotype or TC genotype, so as to aid in identification pig to be lived up to 100kg body weight
The body thickness of backfat：The pig of TT genotype is individual more than the pig of CC genotype up to the 100kg body weight live bodies thickness of backfat, CC bases
Because the pig of type is individual more than the pig of TC genotype up to the 100kg body weight live bodies thickness of backfat；
TT genotype be pig MEG3 genes First Intron the 51st nucleotides be T homozygote；
CC genotype be pig MEG3 genes First Intron the 51st nucleotides be A homozygote；
TC genotype is that the nucleotides of pig MEG3 genes First Intron the 51st is T and A heterozygote；
The nucleotide sequence of MEG3 gene First Introns is as shown in sequence 2 in sequence table.
Claims (8)
- Identified or auxiliary identification pig reaches the method for 100kg body weight live body back fat thickness 1. a kind of, be following (1) and/ Or (2) and/or (3) and/or (4)：(1) it is to detect that the genotype of pig individual is TT genotype or AA genotype or TA genotype, according to The genotype of the pig individual determines that the pig individual reaches 100kg body weight live body thickness of backfat sizes：The pig of AA genotype Individual reaches 100kg up to pig individual of the 100kg body weight live bodies thickness of backfat more than TT genotype, the pig individual of TT genotype Pig individual of the body weight live body thickness of backfat more than TA genotype；The TT genotype is the homozygote that the 460th deoxyribonucleotide of MEG3 genes Second Exon is T；The TA genotype be the 460th deoxyribonucleotide of MEG3 genes Second Exon be T and A it is miscellaneous It is fit；The AA genotype is the homozygote that the 460th deoxyribonucleotide of MEG3 genes Second Exon is A；(2) it is to detect that the genotype of pig individual is AA genotype or GG genotype or AG genotype, according to The genotype of the pig individual determines that the pig individual reaches 100kg body weight live body thickness of backfat sizes：The pig of AA genotype Individual reaches 100kg up to pig individual of the 100kg body weight live bodies thickness of backfat more than GG genotype, the pig individual of GG genotype Pig individual of the body weight live body thickness of backfat more than AG genotype；The AA genotype is the homozygote that the 847th deoxyribonucleotide of MEG3 genes Second Exon is A；The AG genotype be the 847th deoxyribonucleotide of MEG3 genes Second Exon be A and G it is miscellaneous It is fit；The GG genotype is the homozygote that the 847th deoxyribonucleotide of MEG3 genes Second Exon is G；(3) it is to detect that the genotype of pig individual is AA genotype or GG genotype or AG genotype, according to The genotype of the pig individual determines that the pig individual reaches 100kg body weight live body thickness of backfat sizes：The pig of AA genotype Individual reaches 100kg up to pig individual of the 100kg body weight live bodies thickness of backfat more than GG genotype, the pig individual of GG genotype Pig individual of the body weight live body thickness of backfat more than AG genotype；The AA genotype is the homozygosis that the 1158th deoxyribonucleotide of MEG3 genes Second Exon is A Body；The AG genotype is that the 1158th deoxyribonucleotide of MEG3 genes Second Exon is A and G Heterozygote；The GG genotype is the homozygosis that the 1158th deoxyribonucleotide of MEG3 genes Second Exon is G Body；(4) it is to detect that the genotype of pig individual is TT genotype or CC genotype or TC genotype, according to The genotype of the pig individual determines that the pig individual reaches 100kg body weight live body thickness of backfat sizes：The pig of TT genotype Body reaches 100kg bodies up to pig individual of the 100kg body weight live bodies thickness of backfat more than CC genotype, the pig individual of CC genotype Pig individual of the taskwork body thickness of backfat more than TC genotype；The TT genotype is the homozygote that the 51st deoxyribonucleotide of MEG3 genes First Intron is T；The TC genotype be the 51st deoxyribonucleotide of MEG3 genes First Intron be T and C it is miscellaneous It is fit；The CC genotype is the homozygote that the 51st deoxyribonucleotide of MEG3 genes First Intron is C.
- 2. according to the method described in claim 1, it is characterised in that：In (1), the genotype of the detection pig individual is AA genotype or GG genotype or AG bases Because the method for type is following A) or B)：A) direct Sequencing；B the pcr amplification product containing the 460th deoxyribonucleotide of MEG3 genes Second Exon) is sequenced；In (2), the genotype of the detection pig individual is AA genotype or GG genotype or AG bases Because the method for type is following C) or D)：C) direct Sequencing；D the pcr amplification product containing the 1137th deoxyribonucleotide of MEG3 genes Second Exon) is sequenced；In (3), the genotype of the detection pig individual is AA genotype or GG genotype or AG bases Because the method for type is following E) or F)：E) direct Sequencing；F the pcr amplification product containing the 1158th deoxyribonucleotide of MEG3 genes Second Exon) is sequenced；In (4), the genotype of the detection pig individual is TT genotype or CC genotype or TC bases Because the method for type is following G) or H)：G) direct Sequencing；H the pcr amplification product containing the 51st deoxyribonucleotide of MEG3 genes First Intron) is sequenced；The B), D) and F) in pcr amplification product used in primer for it is following 1) or 2)：1) as the single stranded DNA shown in sequence 4 in the single strand dna shown in sequence in sequence table 3 and sequence table The primer pair A of molecular composition；2) primer pair being made up of the single strand dna shown in the single strand dna and sequence B shown in sequence A C；The sequence A has phase to be deleted sequence 3 or increasing or change one or several nucleotides with sequence 3 The nucleotides of congenerous；The sequence B has phase to be deleted sequence 4 or increasing or change one or several nucleotides with sequence 4 The nucleotides of congenerous；The H) in pcr amplification product used in primer for it is following 3) or 4)：3) as the single stranded DNA shown in sequence 6 in the single strand dna shown in sequence in sequence table 5 and sequence table The primer pair B of molecular composition；4) primer pair being made up of the single strand dna shown in the single strand dna and sequence D shown in sequence C D；The sequence C has phase to be deleted sequence 5 or increasing or change one or several nucleotides with sequence 5 The nucleotides of congenerous；The sequence D has phase to be deleted sequence 6 or increasing or change one or several nucleotides with sequence 6 The nucleotides of congenerous.
- 3. detect the 460th deoxyribonucleotide and/or the 1137th of the MEG3 gene Second Exons of pig individual The material of the genotype of position deoxyribonucleotide and/or the 1158th deoxyribonucleotide detects pig individual Identification pig is being identified or aided in the material of the genotype of 51st deoxyribonucleotide of MEG3 gene First Introns Individual reaches the application in the 100kg body weight live body thickness of backfats；Or the 460th deoxyribonucleotide and/or the 1137th of the MEG3 gene Second Exons of detection pig individual The material of the genotype of position deoxyribonucleotide and/or the 1158th deoxyribonucleotide detects pig individual The material of the genotype of 51st deoxyribonucleotide of MEG3 gene First Introns is preparing identification or auxiliary mirror Pig individual is determined up to the application in the product of the 100kg body weight live body thickness of backfats.
- 4. detect the 460th deoxyribonucleotide and/or the 1137th of the MEG3 gene Second Exons of pig individual The material of the genotype of position deoxyribonucleotide and/or the 1158th deoxyribonucleotide detects pig individual Application of the material of the genotype of 51st deoxyribonucleotide of MEG3 gene First Introns in pig breeding；Or the 460th deoxyribonucleotide and/or the 1137th of the MEG3 gene Second Exons of detection pig individual The material of the genotype of position deoxyribonucleotide and/or the 1158th deoxyribonucleotide detects pig individual The material of the genotype of 51st deoxyribonucleotide of MEG3 gene First Introns is preparing the product of pig breeding In application.
- 5. detect the 460th deoxyribonucleotide and/or the 1137th of the MEG3 gene Second Exons of pig individual The material of the genotype of position deoxyribonucleotide and/or the 1158th deoxyribonucleotide detects pig individual The material of the genotype of 51st deoxyribonucleotide of MEG3 gene First Introns seed selection lean meat percentage it is high and/ Or skeletal muscle content height and/or pig reach the application in the short boar of the 100kg body weight live body thickness of backfats；Or the 460th deoxyribonucleotide and/or the 1137th of the MEG3 gene Second Exons of detection pig individual The material of the genotype of position deoxyribonucleotide and/or the 1158th deoxyribonucleotide detects pig individual The material of the genotype of 51st deoxyribonucleotide of MEG3 gene First Introns is preparing seed selection lean meat percentage height And/or skeletal muscle content height and/or pig reach the application in the product of the short boar of the 100kg body weight live body thickness of backfats.
- 6. a kind of identify the product that pig individual reaches the 100kg body weight live body thickness of backfats, for the MEG3 bases of detection pig individual Because of the 460th deoxyribonucleotide and/or the 1137th deoxyribonucleotide and/or the 1158th of Second Exon The 51st of the material of the genotype of position deoxyribonucleotide or the MEG3 gene First Introns of detection pig individual The material of the genotype of deoxyribonucleotide.
- 7. according to any described application in claim 3-5 or the product described in claim 6, it is characterised in that： It is described detection pig individual MEG3 gene Second Exons the 460th deoxyribonucleotide and/or the 1137th The material of the genotype of deoxyribonucleotide and/or the 1158th deoxyribonucleotide detects pig individual The material of the genotype of 51st deoxyribonucleotide of MEG3 gene First Introns is following X1)-X6)：X1) as the single stranded DNA shown in sequence 4 in the single strand dna shown in sequence in sequence table 3 and sequence table The primer pair A of molecular composition；X2) as the single stranded DNA shown in sequence 6 in the single strand dna shown in sequence in sequence table 5 and sequence table The primer pair B of molecular composition；X3) the primer being made up of the single strand dna shown in the single strand dna and sequence B shown in sequence A To C；The sequence A has phase to be deleted sequence 3 or increasing or change one or several nucleotides with sequence 3 The nucleotides of congenerous；The sequence B has phase to be deleted sequence 4 or increasing or change one or several nucleotides with sequence 4 The nucleotides of congenerous；X4) the primer being made up of the single strand dna shown in the single strand dna and sequence D shown in sequence C To D；The sequence C has phase to be deleted sequence 5 or increasing or change one or several nucleotides with sequence 5 The nucleotides of congenerous；The sequence D has phase to be deleted sequence 6 or increasing or change one or several nucleotides with sequence 6 The nucleotides of congenerous；X5 X1) is contained) the primer pair A or X2) the primer pair B or X3) the primer pair C or X4) The PCR reagent of the primer pair D；X6 X1) is contained) the primer pair A or X2) the primer pair B or X3) the primer pair C or X4) The primer pair D or X5) PCR reagent kit.
- 8. a kind of seed selection lean meat percentage is high and/or skeletal muscle content is high and/or pig reaches the short kind of the 100kg body weight live body thickness of backfats The method of pig, including the pig of selection TC genotype and/or TA genotype and/or AG genotype carry out breeding；The TA genotype be the 460th deoxyribonucleotide of MEG3 genes Second Exon be T and A it is miscellaneous It is fit；The TC genotype be the 51st deoxyribonucleotide of MEG3 genes First Intron be T and C it is miscellaneous It is fit；The AG genotype is the 1137th deoxyribonucleotide and/or the 1158th of MEG3 genes Second Exon The heterozygote that position deoxyribonucleotide is A and G.
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| CN112126691A (en) * | 2020-10-15 | 2020-12-25 | 上海新农科技股份有限公司 | Method, primer, kit and application for auxiliary detection of pig backfat thickness character by utilizing pig No. 2 chromosome gene polymorphic site |
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| CN108359733A (en) * | 2017-12-08 | 2018-08-03 | 中国农业科学院北京畜牧兽医研究所 | A method of the auxiliary detection pig 100kg thickness of backfats |
| CN108359733B (en) * | 2017-12-08 | 2021-04-02 | 中国农业科学院北京畜牧兽医研究所 | Method for auxiliary detection of 100kg backfat thickness of pig |
| CN112126691A (en) * | 2020-10-15 | 2020-12-25 | 上海新农科技股份有限公司 | Method, primer, kit and application for auxiliary detection of pig backfat thickness character by utilizing pig No. 2 chromosome gene polymorphic site |
| CN112126691B (en) * | 2020-10-15 | 2024-04-26 | 上海新农科技股份有限公司 | Method, primer, kit and application for auxiliary detection of pig backfat thickness character by using pig chromosome 2 gene polymorphism site |
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