CN102220314A - Method for auxiliary identification of chicken populations with different weight traits - Google Patents
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Abstract
The invention discloses a method for auxiliary identification of chicken populations with different weight traits. The invention provides a primer pair formed by DNA shown by a sequence 3 in a sequence table and DNA shown by a sequence 4 in the sequence table. The primer pair can be applied in the auxiliary identification of the chicken populations with the different weight traits. The method comprises the following steps: respectively taking gene group DNA of each chicken to be identified as a moldboard; carrying out PCR (Polymerase Chain Reaction) amplification by the primer pair formed by the DNA shown by the sequence 3 in the sequence table and the DNA shown by the sequence 4 in the sequence table, and carrying out enzyme fragmentation to a PCR product by restriction enzyme Hhal; and identifying the chicken to be identified to be BB if two DNA segments can be obtained and identifying the chicken to be identified to be AB if three DNA segments are obtained, wherein the average weight of the BB gene type chicken population is statistically lower than the average weight of the AB gene type chicken population. The invention finds that the SNP (Single Nucleotide Polymorphism) in the SCTGF (Similar Connective Tissue Growth Factor) gene of the chicken is remarkably related with the weight body, and can be used as a genetic marker of the chicken. The method can be used for carrying out auxiliary breeding by a molecule biological means.
Description
Technical field
The present invention relates to the method that assistant identification has the chicken colony of different weight characters.
Background technology
SNP (single nucleotide polymorphism single nucleotide polymorphism) mainly is meant on genomic level, and the polymorphism by the caused dna sequence dna of variation of single Nucleotide comprises conversion, transversion, insertion or disappearance.Densely distributed SNP in genome because of it carries huge genetic information amount, is considered to the best genetic marker of application prospect, and the detection of human SNP mark has become the important means of gene diagnosis.The research of the genetic map of SNP mark has huge promoter action to marker assisted selection, and the gene of a lot of important characters is located in the agricultural animal.
Some existing mature technologies are often adopted in the detection of SNP, as dna sequencing (Sanger sequencing and tetra-sodium sequencing), single strand conformation polymorphism (SSCP), restriction fragment length polymorphism (RFLP), allele specific oligonucleotide oligonucleotide hybridization (ASO), oligonucleotide special be connected, technique means such as DNA chip and Taqman probe.
(Bradham DM such as Bradham in 1991, Igarashi A, Potter RL, et al.Connectivetissue growth factor:a cysteine-rich mitogen secreted by human vascularendothelial cells is related to the SRC-induced immediate early gene productCEF-10[J] .J Cell Biol, 1991,114 (6): 1285.) found first human Connective Tissue Growth Factor (connective tissue growth factor, CTGF).CTGF is mainly by mesenchymal cell synthesis secretions such as skin flbroblast, mesangial cell, hepatic stellate cell, lung fibroblast, vascular smooth muscle cell.CTGF is the member in CCN (abbreviation of CTGF, Cef10/cyr61 and the Nov) family.This family comprises Nov, Xnov and the human CTGF that the Africa xenopus inoblast produces that expresses in Cyr61, the Fisp-12 that the 3T3 cell produces after being subjected to serum stimulation through inducing generation of the Cef10 of chick embryo fibroblast through inducing generation, 3T3 cell, the chicken nephrocyte of growing up at present.They all belong to immediate early gene (immediate early gene), participate in the regulation and control of somatomedin cell growth differentiation directly.Their common structure characteristics be similarity with very high sequence homology and sequential structure (Bork P.The modular architecture of a new family of growth regula2torsrelated to connective tissue growth factor[J] .FEBS Lett, 1993,327 (2): 125.).
CTGF plays an important role in organism, a plurality of pathways metabolisms have been participated in, mainly bringing into play effect (Liu Jianyi in the following areas, Li Shirong record refined emerging Connective Tissue Growth Factor and biological action ChineseJournal of Practical Aesthetic and Plastic Surgery thereof, 2004.Vol.15No.1):
(1) quickening DNA synthesizes, promotes cell proliferation: Bradham in 1991 etc. find that at first human CTGF has the mitogenesis effect to the 3T3 cell.
(2) regulating extracellular matrix expression of gene: CTGF can stimulate fibroblast proliferation, extracellular matrix generation, granulation tissue to form and fibrosis.
(3) mediated cell sticks: (Chen CC such as Chen, Chen N, Lau L F.The angiogenic factorsCyr61and connective tissue growth factor induce adhesive signaling in primaryhuman skin fibroblasts[J] .J Biol Chem, 2001,276 (13): 10443.) discover that CTGF can mediate fibroblastic sticking, its process must have the participation of integrating plain and heparan sulfate proteoglycan.
(4) irritation cell migration: discoveries such as Bradham, CTGF can stimulate the migration of the 3T3 cell of vitro culture.(Fan WH such as Fan, Pech M, Karnovsky MJ.Connective tissue growth factor (CTGF) stimulates vascular smooth muscle cell growth and migra2tion in vitro[J] .Eur J Cell Biol, 2000,79 (12): 915.) find, if the CTGF over-expression, the growth rate of vascular smooth muscle cell and mobility all will have remarkable increase.
(5) promote new vessel to form: (Babic AM such as Babic, Chen CC, Lau L F.Fisp12/ mouseconnective tissue growth factor mediates endothelial cell adhesion andmigration through integri
(6) promote cartilage generation and skeleton development: CTGF can promote chondrocyte and osteoblastic proliferation and differentiation, (Nakanishi T such as Nakanishi, Kimura Y, Tamura T, et al.Cloning of a mRNApreferentially expressed in chondrocytes by differential display-PCR from ahuman chondrocytic cell line that is identical with con2nective tissue growthfactor (CTGF) mRNA[J] .Biochem Biophys Res Commun, 1997,234 (1): 206.) find that expressing excessively of CTGFmRNA can promote chondrocyte's strain HCS-2/8 cell proliferation, and promote cartilage to assemble proteoglycan secretion and the expression of X collagen type, and the X Collagen Type VI is loose band phenotype of chondrocytes feature.
(7) promote cell survival, avoid apoptosis: discovery such as Babic place endotheliocyte that the layer that does not contain somatomedin is glutinous to be connected in the protein culture medium (promptly under apoptosis-induced situation), and the CTGF of adding mouse can promote the survival of endotheliocyte to avoid apoptosis.
Body weight index as growth traits is one of economic characters important during livestock industry is produced always, also is the important indicator in the fryer breed breeding.Compare with nineteen fifty-seven, the carcass output of the fryer strain in the present age of calendar year 2001 has improved 6 times, and wherein 85%-90% will be given the credit to the genetics effect, has only 10%-15% to be because the improvement of feed nutrition level causes.The research of poultry genetic marker assistant breeding is deepening continuously, and a large amount of proterties has been determined the candidate region on hereditary level.
There is abundant native chicken breed resource in China, is the problem that presses for solution to its assessment, protection and seed selection how.
Summary of the invention
The purpose of this invention is to provide the method that assistant identification has the chicken colony of different weight characters.
It is right to the invention provides the primer that DNA forms shown in the sequence 4 of DNA shown in the sequence 3 of sequence table and sequence table.
Described primer has the chicken colony of different weight characters to can be applicable to assistant identification.
Described primer is to also can be applicable to prepare the test kit that assistant identification has the chicken colony of different weight characters.
The present invention protects the right assistant identification that contains described primer to have the test kit of the chicken colony of different weight characters simultaneously.
Assistant identification provided by the invention has the method for the chicken colony of different weight characters, comprise the steps: that respectively the genomic dna with every chicken to be measured is a template, the primer of forming with DNA shown in the sequence 4 of DNA shown in the sequence 3 of sequence table and sequence table is cut the PCR product to carrying out pcr amplification with restriction enzyme HhaI enzyme; If obtain two dna fragmentations, the genotype of chicken to be measured is BB; If obtain three dna fragmentations, the genotype of chicken to be measured is AB; The mean body weight of BB genotype chicken colony is lower than the mean body weight of the genotypic chicken of AB colony statistically.
Described two dna fragmentations are specially the dna fragmentation of 45bp and the dna fragmentation of 191bp; Described three dna fragmentations are specially the dna fragmentation of 236bp, the dna fragmentation of 45bp and the dna fragmentation of 191bp.
A kind of assistant identification provided by the invention has the method for the chicken colony of different weight characters, comprises the steps: to detect respectively dna fragmentation shown in the sequence 2 of dna fragmentation shown in the sequence 1 that whether has sequence table in the genomic dna of every chicken to be measured and sequence table; If chicken to be measured is the homozygote of dna fragmentation shown in the sequence 1, its genotype is BB; If chicken to be measured is the heterozygote of dna fragmentation shown in sequence 1 and 2, its genotype is AB; The mean body weight of BB genotype chicken colony is lower than the mean body weight of the genotypic chicken of AB colony statistically.
More than in arbitrary described method, also comprise the step of selecting chicken to be measured; Described chicken to be measured is the chicken of following two colonies: all have dna fragmentation shown in the sequence 2 of dna fragmentation shown in the sequence 1 of sequence table and sequence table in colony in every chicken genome to be measured simultaneously; All have dna fragmentation shown in the sequence 2 of sequence table in another colony in every chicken genome to be measured and do not have dna fragmentation shown in the sequence 1.
More than in arbitrary described method, described chicken to be measured specifically can be the chicken of CAU resource family, the F2 that more specifically can be CAU resource family is for individuality.Described chicken to be measured specifically can be AB or the genotypic chicken of BB.
More than in arbitrary described method, described weight character specifically can be nascent body weight, 1 age in week body weight, 3 age in week body weight, 4 age in week body weight, 5 age in week body weight, 6 age in week body weight or 7 age in week body weight.
Described method, described primer to or described test kit can be applicable to a breed of chicken.
Genomic dna with chicken to be measured is a template, and the primer of forming with DNA shown in the sequence 4 of DNA shown in the sequence 3 of sequence table and sequence table is to carrying out pcr amplification, and the PCR product that obtains is the dna fragmentation shown in the sequence 2 of the sequence 1 of sequence table or sequence table.Only there is the difference of a Nucleotide in dna fragmentation shown in sequence 1 and the sequence 2, and dna fragmentation shown in the sequence 1 is C from the 191st Nucleotide of 5 ' end, and dna fragmentation shown in the sequence 2 is T from the 191st Nucleotide of 5 ' end.The present invention finds that this SNP (the class CTGF gene of chicken) and body weight have the characteristic of significant correlation, can be used as the genetic marker of chicken.
Method provided by the invention and product can carry out assistant breeding, and assisting sifting fast and accurately has early screening, saves time, with low cost, advantage of high accuracy.
Description of drawings
Fig. 1 is a PCR product electrophoretogram; M:100bp Marker; 1-12 swimming lane: 12 Different Individual of CAU resource family; Ck: blank.
Fig. 2 cuts the rear electrophoresis collection of illustrative plates for PCR product enzyme; M:100bp Marker; Ck: blank.
Embodiment
Following embodiment is convenient to understand better the present invention, but does not limit the present invention.Experimental technique among the following embodiment if no special instructions, is ordinary method.Used test materials among the following embodiment if no special instructions, is to buy from routine biochemistry reagent shop and obtains.% among the following embodiment if no special instructions, is the quality percentage composition.Quantitative test in following examples all is provided with repeated experiments three times, results averaged.
CAU resource family: be used to study the experimental population that the location influence chicken important economical trait QTL, seeks clone's major gene by what the Li Ning of China Agricultural University professor seminar set up in 1998, the parent is French star's fryer and calm and peaceful plumage Gallus Domesticus; The source is China Agricultural University experiment pasture; CAU resource family reference: Deng, X.M., Li, J.Y., Li, N., and Wu, C.X. (2001) .[Genetic analysis of important growth traitbased on F-2resource population in chicken] .Yi Chuan Xue Bao 28,801-807.; China Agricultural University guarantees to provide to the public.
The discovery of embodiment 1, SNP and body weight dependency
By the comparative genomics analysis, on No. 23 karyomit(e)s of chicken, found people's CTGF analogue (similarto connective tissue growth factor, LOC419598), with with the gene order-checking of chicken BLAT as a result, find that SCTGF is positioned at No. 23 karyomit(e)s (Chr23:2887664-2896948) of chicken, be made up of 4 exons, transcript length is 966bp, 322 amino acid of encoding.
Growth traits is at present about one of maximum content of the QTL research of chicken.In CAU resource family, measured F2 for the individuality body weight in 1 to 12 age in week, located a series of QTL that influence the chicken growth according to scanning result.Wherein it is worth noting most and near No. 23 chromosomal marker ADL0262 of chicken, located a QTL who influences chicken birth weight and early growth (1 to 7 age in week) body weight, order-checking information according to chicken, analyzing this regional gene can find some genes to further investigate as the candidate gene that influences chicken early growth body weight, (connectivetissue growth factor, CTGF) (NC_006110) is exactly candidate gene at first to Connective Tissue Growth Factor.
One, gene type assay
Experiment material: the F2 of 215 CAU resource familys is for individuality.
1, the extraction of genomic dna
Chicken is the wing venous blood collection during 12 ages in week, and the back cracking is handled in anti-freezing, and behind protease K digesting, with the imitative extracting of phenol, TE dissolves-20 ℃ of preservations.
2, pcr amplification
The genomic dna that extracts with step 1 is a template, with primer CTGFex3H is carried out pcr amplification.
CTGFex3H: forward primer: 5 '-AAATCGAACCATCGGGATTA-3 ';
Reverse primer: 5 '-GGTCTGCACCAAGCAGTTCT-3 '.
PCR reaction system (15 μ l): template 2 μ l (40ng genomic dna), amplification buffer (10 * buffer) 1.5 μ l, dNTP 1.2 μ l, forward primer 0.8pm/ul, reverse primer 0.8pm/ul, archaeal dna polymerase (TaqE) 0.3 μ l adds water and mends to 15 μ l.
PCR reaction conditions: 94 ℃, 5min; 94 ℃, 30sec, 60 ℃, 30sec, 72 ℃, 90sec, 40 circulations; 72 ℃, 7min.
The PCR product carries out 2% sepharose (see figure 1), reclaims the dna fragmentation of 236bp.
3, enzyme is cut
Pcr amplification product carries out enzyme with HhaI and cuts (37 ℃ of 5h).Enzyme is cut system (20 μ l): PCR product 10 μ l, and 10 times of damping fluid 2 μ l, 2U HhaI (TaKaRa company), water is mended to 20 μ l.
Enzyme is cut product and is detected (see figure 2) with 4% sepharose.
Produce three kinds of banding patterns:
First kind of banding pattern: have only a band, be 236bp; With genotype called after AA;
Second kind of banding pattern: two bands are arranged, be respectively 45bp and 191bp; With genotype called after BB, the 45bp fragment is less, fails to show, so Fig. 2 shows the band of a 191bp;
The third banding pattern: three bands are arranged, be respectively 45bp, 191bp and 236bp; With genotype called after AB, the 45bp fragment is less, fails to show, so Fig. 2 shows two bands of 191bp and 236bp.
Statistics such as table 1.
The table 1CTGF gene PCR-distribution of HhaI-RFLP identified gene type in F2 colony
From table 1 result, the BB type is the preponderant genotype in the used colony of experiment, and ratio reaches 80%.
Two, sequence verification
PCR product with each sample checks order respectively.The result shows, in the chicken of all kinds: the AA genotype is the homozygote (homozygote of 2448T) of the DNA shown in the sequence 2 of sequence table; The BB genotype is the homozygote (homozygote of 2448C) of the DNA shown in the sequence 1 of sequence table; The AB genotype is the heterozygote (heterozygote of 2448T and 2448C) of the DNA shown in the sequence 1 of DNA shown in the sequence 2 of sequence table and sequence table.
Contrast CTGF gene (NC_006110) is analyzed: the SNP at 2448 places of exon 3 is same sense mutation, because when this SNP sports T by C, can reduce by a HhaI restriction enzyme site, thereby can adopt the variation situation of the method detection 2448C-T of PCR-HhaI-RFLP; Can obtain containing the part dna fragmentation (236bp) of exon 3 with the amplification of CTGFex3H primer PCR, the amplified production of 236bp is after the HhaI enzyme is cut, produce three kinds of banding patterns, corresponding different respectively genotype: 236bp (AA), 45+191bp (BB) and 45+191+236bp (AB).
Three, association analysis
The PROC GLM process of employing SAS (8.2 editions) statistical analysis software bag is carried out statistical study and according to the least square analytical model genotype and the growth traits that supply examination chicken group is carried out the variance statistical study.
Statistic analysis models:
Y
ijkl＝U+F
i+S
j+R
k+e
ijkl
Y
IjklmIndividual phenotypic character (body weight) observed value;
U is the average of colony;
F
iBe the effect value of i family to phenotypic character (body weight);
S
jBe the effect value of sex to phenotypic character (body weight);
R
kBe the effect value of different batches to phenotypic character (body weight);
e
IjklBe random residual effect corresponding to observed value.
Analytical results sees Table 2.
The variance analysis of the polymorphism and chicken of table 2CTGF gene PCR-HhaI-RFLP body weight in each in all age
| AA | AB | BB | |
| W0 (coming into being) | 30.03±1.20 ab | 31.79±0.54 ab | 29.75±0.28 b |
| W1 (1 age in week) | 73.10±4.51 ab | 70.90±2.03 a | 64.51±1.05 b |
| W2 (2 age in week) | 153.72±9.06 a | 144.66±4.09 a | 135.80±2.12 a |
| W3 (3 age in week) | 251.97±13.94 ab | 241.65±6.29 a | 225.16±3.26 b |
| W4 (4 age in week) | 382.03±23.32 ab | 366.78±10.52 a | 341.73±5.45 b |
| W5 (5 age in week) | 541.24±34.84 ab | 536.65±15.71 a | 491.53±8.14 b |
| W6 (6 age in week) | 726.64±45.55 ab | 726.77±20.54 a | 669.75±10.65 b |
| W7 (7 age in week) | 942.05±59.83 ab | 918.53±26.98 a | 851.39±13.99 b |
| W8 (8 age in week) | 1076.35±69.91 a | 1121.48±31.53 a | 1053.89±16.34 a |
| W9 (9 age in week) | 1297.92±76.90 a | 1334.59±34.68 a | 1253.35±17.98 a |
| W10 (10 age in week) | 1477.05±85.82 a | 1514.59±38.70 a | 1436.62±20.06 a |
| W11 (11 age in week) | 1600.09±90.95 a | 1645.78±41.02 a | 1587.89±21.26 a |
| W12 (12 age in week) | 1750.22±91.03 a | 1762.45±43.76 a | 1715.40±22.68 a |
Annotate: average comparand parent phase is not remarkable with expression difference, and alphabetical different table differential is different significantly.
Wherein in w0 and w1, the BB type reaches difference extremely significantly (P＜0.01) to the AB type, and from W3 and W7, the BB type reaches significant difference (P＜0.05) to the AB type.
Analytical results shows: each etap AB genotype colony the average body weight average greater than the mean body weight of BB genotype colony, the BB type is the inferior position genotype; In birth heavy (birth weight) with during 1 age in week, body weight analyzed, difference is (P＜0.01) extremely significantly to the AB type for the BB type, the BB type to the AB type from 3 body weight analyses in age in week ages to 7 in week, significant difference (P＜0.05).General 6 weeks of commercial meat bird Time To Market average out to.
The application of embodiment 2, SNP
Chicken (all available from China Agricultural University experiment pasture) to 7 kinds carries out SNP mensuration, 7 kinds respectively: Guangxi three yellow chickens, Beijing fine breed of chicken with thick brownish feathers, Luo Man fryer, Tibetan chicken, the brown laying hen of agricultural university, silk plumage Gallus Domesticus, star's fryer.
1, the extraction of genomic dna
Chicken is the wing venous blood collection during 12 ages in week, and the back cracking is handled in anti-freezing, and behind protease K digesting, with the imitative extracting of phenol, TE dissolves-20 ℃ of preservations.
2, the genotype of each kind is determined
The genomic dna that extracts with step 1 is a template, with primer CTGFex3H is carried out pcr amplification.
CTGFex 3H: forward primer: 5 '-AAATCGAACCATCGGGATTA-3 ';
Reverse primer: 5 '-GGTCTGCACCAAGCAGTTCT-3 '.
PCR reaction system (15 μ l): template 2 μ l (40ng genomic dna), amplification buffer (10 * buffer) 1.5 μ l, dNTP 1.2 μ l, forward primer 0.8pm/ul, reverse primer 0.8pm/ul, archaeal dna polymerase (TaqE) 0.3 μ l adds water and mends to 15 μ l.
PCR reaction conditions: 94 ℃, 5min; 94 ℃, 30sec, 60 ℃, 30sec, 72 ℃, 90sec, 40 circulations; 72 ℃, 7min.
The PCR product carries out 2% sepharose, reclaims the dna fragmentation of 236bp.
Pcr amplification product carries out enzyme with HhaI and cuts (37 ℃ of 5h).Enzyme is cut system (20 μ l): PCR product 10 μ l, and 10 times of damping fluid 2 μ l, 2U HhaI (TaKaRa company), water is mended to 20 μ l.
Enzyme is cut product and is detected with 4% sepharose.
The results are shown in Table 3.
The distribution of polymorphism in different standard-bred poultries of table 3CTGF gene PCR-HhaI-RFLP
The polymorphism of table 4CTGF gene PCR-HhaI-RFLP at home with external chicken kind in distribution
| Total sample number | AA | AB | BB | |
| Domestic chicken kind | 79 | 1 | 5 | 73 |
| External chicken kind | 78 | 20 | 42 | 16 |
Chi square test: x
2=82.82, d
f=3, P＜0.01.
From table 3 and table 4, find out, seven standard-bred poultries detect two allelotrope of A and B equally, are divided into AA, AB, three kinds of genotype of BB, the AB genotypic proportion is bigger in external chicken breed Luo Man fryer and the star's fryer, and the BB genotype is a main body in place of china chicken kind.The AB genotype is for influence the preponderant genotype of the heavy and early growth body weight of chicken birth, and the BB type is the inferior position genotype, so the distribution of the genotype in this site is consistent with the difference of domestic and international chicken kind body weight and growth.
The result shows that the BB homozygous individual has inferior position aspect body weight.This SNP site can be used as a genetic marker, is applied to the molecular genetic marker assisted Selection of chicken growth traits, improves the speed and the accuracy of fryer seed selection.
Sequence table
＜110〉China Agricultural University
＜120〉assistant identification has the method for the chicken colony of different weight characters
<130>CCGNARY102223
<160>4
<210>1
<211>236
<212>DNA
＜213〉chicken (Gallus domestiaus)
<400>1
aaatcgaacc?atcgggatta?tatagaatta?taaagatcac?agaaccatga?aatcacagaa 60
tggttgggtt?ggaagggaac?ttaaagaccc?ccagctttgc?ctcccaccgg?atcaggctgt 120
ccagggccct?atggggtatc?tgaccttcat?gtcttcctcc?tggctcagtt?ttcagggagg 180
atcccgagcg?cgagccggag?ctgaacaacc?tgcaggagaa?ctgcttggtg?cagacc 236
<210>2
<211>236
<212>DNA
＜213〉chicken (Gallus domestiaus)
<400>2
aaatcgaacc?atcgggatta?tatagaatta?taaagatcac?agaaccatga?aatcacagaa 60
tggttgggtt?ggaagggaac?ttaaagaccc?ccagctttgc?ctcccaccgg?atcaggctgt 120
ccagggccct?atggggtatc?tgaccttcat?gtcttcctcc?tggctcagtt?ttcagggagg 180
atcccgagcg?tgagccggag?ctgaacaacc?tgcaggagaa?ctgcttggtg?cagacc 236
<210>3
<211>20
<212>DNA
＜213〉artificial sequence
<220>
<223>
<400>3
aaatcgaacc?atcgggatta 20
<210>4
<211>20
<212>DNA
＜213〉artificial sequence
<220>
<223>
<400>4
ggtctgcacc?aagcagttct 20
Claims (10)
1. the primer of the composition of DNA shown in the sequence 4 of DNA and sequence table shown in the sequence 3 of sequence table is right.
2. the described primer of claim 1 is to the application in the chicken colony that has different weight characters in assistant identification.
3. the described primer of claim 1 is to the application in the test kit of chicken colony that has different weight characters in the preparation assistant identification.
4. an assistant identification has the test kit of the chicken colony of different weight characters, comprises that the described primer of claim 1 is right.
5. an assistant identification has the method for the chicken colony of different weight characters, comprises the steps: that respectively the genomic dna with every chicken to be measured is a template, to carrying out pcr amplification, cuts PCR product with restriction enzyme HhaI enzyme with the described primer of claim 1; If obtain two dna fragmentations, the genotype of chicken to be measured is BB; If obtain three dna fragmentations, the genotype of chicken to be measured is AB; The mean body weight of BB genotype chicken colony is lower than the mean body weight of the genotypic chicken of AB colony statistically.
6. method as claimed in claim 5 is characterized in that: described two dna fragmentations are the dna fragmentation of 45bp and the dna fragmentation of 191bp; The dna fragmentation that described three dna fragmentations are 236bp, the dna fragmentation of 45bp and the dna fragmentation of 191bp.
7. an assistant identification has the method for the chicken colony of different weight characters, comprises the steps: to detect respectively dna fragmentation shown in the sequence 2 of dna fragmentation shown in the sequence 1 that whether has sequence table in the genomic dna of every chicken to be measured and sequence table; If chicken to be measured is the homozygote of dna fragmentation shown in the sequence 1, its genotype is BB; If chicken to be measured is the heterozygote of dna fragmentation shown in sequence 1 and 2, its genotype is AB; The mean body weight of BB genotype chicken colony is lower than the mean body weight of the genotypic chicken of AB colony statistically.
8. as arbitrary described method in the claim 5 to 7, it is characterized in that: described method also comprises the step of selecting chicken to be measured; Described chicken to be measured is the chicken of following two colonies: all have dna fragmentation shown in the sequence 2 of dna fragmentation shown in the sequence 1 of sequence table and sequence table in colony in every chicken genome to be measured simultaneously; All have dna fragmentation shown in the sequence 2 of sequence table in another colony in every chicken genome to be measured and do not have dna fragmentation shown in the sequence 1.
9. as arbitrary described method in the claim 5 to 8, it is characterized in that: described chicken to be measured is the chicken of CAU resource family, and the F2 that is preferably CAU resource family is for individuality; Described weight character for nascent body weight, 1 age in week body weight, 3 age in week body weight, 4 age in week body weight, 5 age in week body weight, 6 age in week body weight or 7 age in week body weight.
10. the described primer of claim 1 is right, the application of arbitrary described method in a breed of chicken in described test kit of claim 4 or the claim 5 to 9.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103276090A (en) * | 2013-05-30 | 2013-09-04 | 扬州大学 | Jinghai yellow chicken 16-week weight molecular genetic marker and application thereof |
| CN103710427A (en) * | 2013-09-27 | 2014-04-09 | 河南农业大学 | Single nucleotide polymorphism, detection method and application of chicken gene |
| CN109402285A (en) * | 2018-11-09 | 2019-03-01 | 北京林业大学 | A kind of classifying method based on complete genome DNA methylation sites genotype |
| CN110951889A (en) * | 2018-09-26 | 2020-04-03 | 中国农业大学 | Haplotype molecular marker related to chicken body weight and application thereof |
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2010
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Non-Patent Citations (2)
| Title |
|---|
| 《Animal Genetics》 20060730 Y. Gao et al 《A genome scan for quantitative trait loci associated with body weight at different developmental stages》 276-278 1-10 第37卷, 第3期 * |
| 《Animal Genetics》 20100228 Y. Gao1 et al 《Identification of quantitative trait loci for shank length and growth at different development stages in》 全文 1-10 第41卷, 第1期 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103276090A (en) * | 2013-05-30 | 2013-09-04 | 扬州大学 | Jinghai yellow chicken 16-week weight molecular genetic marker and application thereof |
| CN103276090B (en) * | 2013-05-30 | 2013-12-11 | 扬州大学 | Jinghai yellow chicken 16-week weight molecular genetic marker and application thereof |
| CN103710427A (en) * | 2013-09-27 | 2014-04-09 | 河南农业大学 | Single nucleotide polymorphism, detection method and application of chicken gene |
| CN103710427B (en) * | 2013-09-27 | 2015-06-17 | 河南农业大学 | Single nucleotide polymorphism, detection method and application of chicken gene |
| CN110951889A (en) * | 2018-09-26 | 2020-04-03 | 中国农业大学 | Haplotype molecular marker related to chicken body weight and application thereof |
| CN110951889B (en) * | 2018-09-26 | 2021-09-21 | 中国农业大学 | Haplotype molecular marker related to chicken body weight and application thereof |
| CN109402285A (en) * | 2018-11-09 | 2019-03-01 | 北京林业大学 | A kind of classifying method based on complete genome DNA methylation sites genotype |
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