CN106399526B - The gene type rapid identification method of chicken beard character - Google Patents
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Abstract
The present invention provides the gene type rapid identification method of a breeder beard character.There is the copy number change in location of CNV in the genome according to chicken beard gene in the present invention, designed for identifying the PCR primer combination of chicken beard character gene type, Direct Identification genotype is identified using Standard PCR, electrophoretic techniques and PCR product band brightness, develop a kind of simple, quick, precise Identification chicken beard gene classifying method, it can accelerate the breeding process that breeding has beard feature chicken kind, shorten the beard chicken breeding time.
Description
Technical field
The present invention relates to molecular biology and genetic breeding fields, specifically, the genotype for being related to chicken beard character is fast
Fast identification method.
Background technique
China is that one of the most abundant country of poultry genetic resources, the feather and properties and characteristics diversity of family chicken are rich in the world
Richness, often by the characteristic trait as certain kinds.The beard of chicken be it is a kind of under cheek two sides and chin raw extension growth
Feather type.In China's Local chicken breeds, Huiyang beard chicken, Beijing Fatty Chicken, Silky fowl etc. all have beard character.Favour
Positive beard chicken is China's Local Excellent meat type kind, and be otherwise known as three yellow beard chickens, Huizhou chicken, gantry chicken, imperial hilllock chicken etc., volume
(national Genetic Resources of Domestic Animal committee group compiles Animal Genetic Resources in China will place to the lower beard shape Fang Yu opened in flourishing
The Beijing kind atlas: Chinese agriculture publishing house, 2015.2).Huiyang beard chicken constitution is solid, and chest muscle is flourishing, thin with meat tenderness bone,
The crisp delicious appearance with three yellow beards of skin has won fame both at home and abroad.
The beard character of chicken is the same with the characters such as the rose comb of chicken, pea comb, multiple hat, silk plumage, thread head, frizzled feather and naked neck,
All can directly it reflect in external phenotype to determine kind and for breeding work.At the beginning of the research of beard character starts from eighties of last century, grind
Study carefully personnel by hybrid experiment determine beard be a kind of autosome incomplete dominant lnheritance character (Davenport,
C.B.1906.Inheritance in poultry.Washington,DC:Carnegie Institution
Publication,1906；Serebrovsky,A.S.,Petrov,S.G.1930.On the composition of the
plan of the chromosomes of the domestics hen.J.Exp.Biol.Russia 6:157-179.).So
And (Guo, Y., et al., A Complex is just reported in the accurate positionin in the genome of chicken beard character and parsing recently
Structural Variation on Chromosome 27Leads to the Ectopic Expression of
HOXB8and the Muffs and Beard Phenotype in Chickens.PLoS Genet,2016.12(6):
p.e1006071；The assignment of genes gene mapping and functional study of Guo Ying chicken beard character, 2016, China Agricultural University's doctorate opinion
Text).The research uses whole-genome association (Genome-wide association study, GWAS), linkage analysis
(Linkage analysis), homologous consistency (Identity by descent, IBD) analysis, comparative genomic hybridization hybrid chip
(Array comparative genomic hybridization, array CGH), full-length genome resurvey sequence and expression analysis
Etc. technological means study the reason gene for causing beard character.Result of study shows that beard character is by multiple on No. 27 chromosomes
Miscellaneous structure variation (Structural variation, SV) causes, this structure variation by be located at 1.7Mb on No. 27 chromosomes,
3 copy number variations (Copy number variation, CNV) of the position 3.5Mb and 4.4Mb form.3 CNV with
CNV3, CNV2 and CNV1's is sequentially and serially coupled together, and is inserted into the downstream (Fig. 1) of CNV1.The result is in other chicken beards chicken group
(including Beijing Fatty Chicken, Yangzhou beard chicken, Polish Bantam, Dutch Owl etc.) verifying is associated with completely with beard character in body.This
Labyrinth variation includes seven annotation genes, the expression to the different phase gene that seven genes are developed in beard in section altogether
Situation is detected, the results showed that the continuous dystopy height expression in lower jaw skin histology of HOXBB gene shows it recklessly strongly
Must play a significant role during development of characters.
Since chicken beard character is controlled by a dominant inheritance gene on autosome, cannot be distinguished from phenotype pure
Mould assembly beard gene individual (Mb/Mb) and heterozygosity beard gene are individual (Mb/mb), can only be determined by way of test cross, be
The selection of beard character is made troubles.The positioning of chicken beard gene (Mb) and the research and production for being accredited as this character of chicken beard
Using the condition of providing the foundation.The method that Guo etc. (2016) reports an identification beard gene type is multiple using beard gene
Occur a base mutation (C → T) in miscellaneous structure in duplicate CNV1, establishes a kind of gene based on pyrosequencing point
Type method.However this method experimentation is more, and needs expensive pyrosequencing instrument, is not easy to promote in practical applications and answer
With.
Summary of the invention
The object of the present invention is to provide the gene type rapid identification methods of a breeder beard character.
There is the copy number change in location of CNV according to chicken beard gene in the present invention, with Standard PCR, electrophoresis in the genome
Technology and PCR product band brightness identify Direct Identification genotype, develop a kind of simple, quick, precise Identification chicken beard base
The classifying method of cause.
In order to achieve the object of the present invention, the present invention is based on 3 copy number variations in beard gene structure (CNV1, CNV2 and
CNV3) position is different from non-beard chicken in the genome, provides the PCR primer combination for identifying chicken beard character gene type,
Include:
Forward primer F1:5'-TGGTTGTGTGCCAAGTAGAG-3'(SEQ ID NO.1)；
Forward primer F2:5'-GCTCTTGCTCTTTTTGGTAA-3'(SEQ ID NO.2)；And
Reverse primer R:5'-CTTTTCGTACTGCGTTGTTA-3'(SEQ ID NO.3).
The present invention also provides the kits for being used to identify chicken beard character gene type containing the combination of above-mentioned primer.
The present invention also provides the gene type rapid identification methods of chicken beard character, comprising the following steps:
1) genomic DNA of chicken to be measured is extracted；
2) it using the DNA extracted in step 1) as template, is combined using above-mentioned primer, carries out pcr amplification reaction；
3) PCR product is analyzed.Electrophoresis is carried out to PCR product using electrophoretic techniques；Then Image J software analytical electrophoresis is used
Glue figure calculates the brightness of band and its ratio in each swimming lane, therefore, it is determined that the genotype of chicken to be measured.
Wherein, the forward primer F1 and the reverse primer R partner primer, expand beard gene 207bp sequence；
The forward primer F2 and the reverse primer R partner primer, expand non-beard gene 296bp sequence.
PCR reaction system is calculated as with 20 μ l:
PCR reaction condition are as follows: 95 DEG C 5 minutes；95 DEG C 30 seconds, 53 DEG C 30 seconds, 72 DEG C 20 seconds, totally 27 circulation；72 DEG C 7 points
Clock.
The method that step 3) analyzes PCR product is as follows:
Pcr amplification product is subjected to 2% agarose gel electrophoresis detection, ethidium bromide staining is seen in gel imaging system
PCR amplification band is examined, and is taken pictures.With Image J software analytical electrophoresis glue figure, 296bp band and 207bp in each swimming lane are calculated
The brightness value of band, and calculate the ratio P of 207bp band brightness value Yu 296bp band brightness value；P value is sentenced between 0.7-0.9
It is set to beard chicken genetic homozygous, P value is determined as beard chicken giblets zygote between 0.2-0.5；P value is 0, only amplifies 296bp
Band is then determined as non-beard chicken individuals.
Beard chicken of the present invention includes Huiyang beard chicken, Beijing Fatty Chicken, Silky fowl etc..Non- beard chicken includes white next
Boat chicken etc..
The present invention also provides specific molecular marker relevant to chicken beard character, the molecular labeling is to be located at chicken No. 27
The downstream CNV1 (newly-increased CNV3 and the junction CNV2, Fig. 2) on chromosome, size is the DNA fragmentation of 207bp, for expanding
The primer of molecular labeling is stated as shown in SEQ ID NO.1 and 3.The amplified production of 207bp size is sequenced, DNAMAN is passed through
Software compares, and confirms its (Fig. 3) consistent with the base sequence of the junction CNV2 with CNV3.
The present invention also provides application of the specific molecular marker in identification beard chicken breed.
The present invention further provides application of the specific molecular marker in chicken molecular mark.
The present invention provides the genotype molecule identification method of a breeder beard gene and applications, that is, use polymerase chain
(PCR) and electrophoretic techniques are reacted to detect with the homozygous of chicken beard gene, heterozygous and non-beard chicken, are produced according to PCR
The band number of object glue figure and band brightness carry out the Rapid identification of chicken beard character gene type, and it is special with beard can to accelerate breeding
The breeding process of chicken kind is levied, the beard chicken breeding time is shortened.
Detailed description of the invention
Fig. 1 is that Read depth analyzes CNV and its rearranged form existing for determining No. 27 chromosomes of beard chicken.
Fig. 2 is that chicken beard gene serotype specific primer expands position in the embodiment of the present invention 2.
Fig. 3 is that peak figure and position is sequenced in the amplified production of chicken beard gene 207bp size of the present invention.
Fig. 4 is that chicken beard gene identifies PCR product electrophoresis result in the embodiment of the present invention 2.
Fig. 5 is that Image J software calculates PCR product band brightness schematic diagram in the embodiment of the present invention 2.
Fig. 6 is that chicken beard gene identifies PCR product electrophoresis result in the embodiment of the present invention 3.
Fig. 7 is the sensitivity analysis result of primer in the embodiment of the present invention 4.
Specific embodiment
The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention..Unless otherwise specified, embodiment
According to conventional laboratory conditions, such as Sambrook molecular cloning experiment handbook (Sambrook J&Russell DW,
Molecular Cloning:a Laboratory Manual, 2001), or according to the condition of manufacturer's specification suggestion.
The genotype and its genetic development of 1 test cross of embodiment identification beard chicken
10 beard cocks from Guangdong Jin Zhong agriculture and animal husbandry Science and Technology Co., Ltd., beard chicken national level conservation field, Huiyang
Chicken and China Agricultural University's experiment 150 of pasture it is white come aircraft carrier chicken (being non-beard chicken), match 15 hens according to 1 cock
Mode set up 10 familys and carry out artificial insemination, every hen collects 10~15 pieces of hatching eggs, amounts to about 1800 pieces of hatching eggs and carries out
Hatching.
F1 generation chicken after hatching and nestling all broods, and raises to 9 week old, observes beard character, counts (table according to family
1).Except No. 7 and No. 9 beard cock chickens and white Leghorn first familiar generation chicken have beard and without beard chicken individuals close to 1:1 in addition to, other
8 beard cock chickens and the first familiar generation chicken of white Leghorn all have beard character.
1 beard chicken of table and white Leghorn first familiar generation beard character count
| Cock family number | There is beard | Without beard |
| 1 | 126 | 0 |
| 2 | 145 | 0 |
| 3 | 142 | 0 |
| 4 | 155 | 0 |
| 5 | 113 | 0 |
| 6 | 149 | 0 |
| 7 | 43 | 46 |
| 8 | 113 | 0 |
| 9 | 58 | 58 |
| 10 | 115 | 0 |
Test cross result explanation, for chicken beard character by Dominant gene, the non-beard trait expression of beard character pair is complete
Dominant, in addition to No. 7 and No. 9 cocks are beard gene heterozygous, other 8 are that beard gene is homozygous.
The foundation of the genotype identification method of 2 chicken beard gene of embodiment
1, design of primers:
In the genome according to 3 copy number variations (CNV1, CNV2 and CNV3) of chicken beard character gene labyrinth
The rear end chicken genome C NV3 600bp sequence, the front end CNV2 upstream 600bp sequence and the front end CNV2 are downloaded in the position of distribution respectively
600bp sequence.3 primers: forward primer F1:5'- are separately designed using 5 software of Primer Premier
TGGTTGTGTGCCAAGTAGAG-3', forward primer F2:5'-GCTCTTGCTCTTTTTGGTAA-3', reverse primer R:5'-
CTTTTCGTACTGCGTTGTTA-3'.Design of primers design is as follows: the present invention is in view of identifying chicken beard with least primer
Genotype.Design of primers principle is that primer binding zone section sequence preservative, the prediction annealing temperature of 3 primers be close, 3 primer structures
At 2 amplified production length 200-400bp between and length difference can pass through electrophoresis significant difference.In addition, comprehensively considering
The factors such as primer G/C content, product dimer, hairpin structure, the continuous base of prime end, positive and negative primer annealing temperature, it is final logical
It crosses optimization and relatively determines above 3 primers.3 primers have two amplicons of A and B in beard chicken and non-beard chicken genomic DNA
(Fig. 2), A amplicon length 207bp, B amplicon length 296bp.In non-beard chicken, only B amplicon, product length
296bp.Designed primer is synthesized by bio-engineering corporation, and is diluted to 10 μm of ol/L of concentration.
2, experimental animal:
Acquire the homozygous individual of 2 beard genes determined in embodiment 1 by test cross, 2 beard gene heterozygous
Body and 2 white Leghorn (non-beard chicken) blood.
3, DNA is extracted:
Chicken blood sample gene group DNA is extracted with DNA extraction kit, using NanoDrop2000 ultramicron spectrophotometric
Meter measurement DNA concentration, and use ddH2O is diluted to 60~100 μ g/ml, the template as PCR amplification.
4, PCR amplification condition optimizing:
(1) optimization of annealing temperature
Using the genomic DNA aggregate sample of 6 beard chickens and non-beard chicken as template, PCR system is 20 μ l, wherein 2 × PCR
10 μ l of Mix dosage, genomic DNA template 1 μ l, 10 μm of 0.5 μ L of ol/L forward primer F1,10 μm of 0.5 μ of ol/L forward primer F2
L, 10 μm of 1 μ L of ol/L reverse primer R, finally use ddH2O is supplemented to 20 μ L, mixes to get pcr amplification reaction system.PCR expands
The response procedures of increasing are as follows: 95 DEG C of initial denaturation 5min；The annealing of 8 change of gradients is arranged within the scope of 49-58 DEG C by 95 DEG C of denaturation 30s
Temperature 30s, 72 DEG C of extension 20s, 36 recycle；72 DEG C of extension 7min；Obtain grads PCR product.PCR product with 2% agar
Carbohydrate gum electrophoresis, the results show that obtaining two PCR product bands of 207bp and 296bp, primer band under 8 annealing temperature conditions
It is neat, clear, without miscellaneous band, illustrate that 3 primer multiplex PCR annealing regions of the invention are wider, the present invention selects intermediate
53 DEG C of the gradient temperature optimization annealing temperatures as 3 primers.
(2) optimization of PCR cycle number
PCR amplification is carried out using the DNA of 6 known beard gene genotype individuals as template, PCR reaction system is 20 μ
L, wherein 2 × PCR Mix dosage, 10 μ l, 1 μ l of genomic DNA, 10 μm of 0.5 μ L of ol/L forward primer F1,10 μm of ol/L forward directions
2 0.5 μ L of primers F, 10 μm of 1 μ L of ol/L reverse primer R, finally use ddH2O is supplemented to 20 μ L, mixes to get pcr amplification reaction
System.The response procedures of PCR amplification are as follows: 95 DEG C of initial denaturation 5min；95 DEG C of denaturation 30s, 53 DEG C of annealing 30s, 72 DEG C of extension 20s,
25~35 circulations；72 DEG C of extension 7min；Obtain PCR product.It is carried out 6 times with 25,27,29,31,33,35 circulations respectively
PCR amplification.
The Ago-Gel for being 2% with concentration carries out electrophoresis to the above different recurring number PCR products, and applied sample amount is 8 μ L.It sees
Electrophoresis result is examined, non-beard chicken has the band of 1 296bp；Homozygous and heterozygous beard chicken individuals have 296bp and
2 band of 207bp.Selection is under 296bp band brightness unanimous circumstances, and homozygous individual 207bp band is than heterozygous 207bp
The brighter glue figure (Fig. 4) of band, the PCR cycle number as optimization.Test result discovery, under the conditions of 27 PCR recycled, energy
The difference of enough homozygous and heterozygous from the band brightness of 207bp identification beard gene, the final optimization recurring number for determining PCR
It is 27.
5, electrophoretic band Luminance Analysis:
The picture file that Fig. 4 is opened using Image J software, analyzes the brightness of PCR product band in each swimming lane, passes through
Identify the calculated by peak area brightness value (Fig. 5) of band.Then the ratio P (table 2) of the brightness of 207bp and 296bp band is calculated, really
Determine the homozygous P value range with heterozygous of beard gene.It is analyzed, determines that the P value of homozygous individual is more than or equal to 0.7, heterozygosis
The P value of type individual is less than 0.5.
2 PCR product band luminance quantization value of table
| Genotype | Sample number | 296bp band brightness | 207bp band brightness | P value |
| Non- beard | 1 | 57.3 | - | 0 |
| Non- beard | 2 | 55.3 | - | 0 |
| Beard gene heterozygous | 3 | 142.2 | 53.4 | 0.38 |
| Beard gene heterozygous | 4 | 114.9 | 49.5 | 0.43 |
| Beard gene is homozygous | 5 | 96.3 | 77.0 | 0.80 |
| Beard gene is homozygous | 6 | 103.7 | 78.2 | 0.75 |
The verifying of the genotype identification method of 3 chicken beard gene of embodiment
From purebred Huiyang beard chicken, beard chicken and white Leghorn first familiar generation, pure in China Agricultural University's practice pasture
The homozygous individual of 7 beard genes, 7 beard gene heterozygous individuals and 6 non-beard chickens are acquired in Zhong Bai Leghorn group
Individual amounts to 20 individual blood samples.All blood samples conceal genotype information, random number.
DNA extraction, PCR product acquisition, agarose gel electrophoresis, electrophoretic band are carried out according to the method that embodiment 2 is established
Luminance Analysis, electrophoresis result are shown in that Fig. 6, band brightness identification and ratio are shown in Table 3.
There was only the band of a 296bp in swimming lane 4,6,10,13,14,18, is directly determined as non-beard chicken.Swimming lane 1,2,
3, there are 2 bands of 296bp and 207bp in 15,16,17,19, and the difference in brightness of 2 bands is little, P value, which is all larger than, to be equal to
0.7, therefore, it is determined that homozygous for beard gene.There are 2 bands of 296bp and 207bp in swimming lane 5,7,8,9,11,12,20, and
Clearly visible 207bp band brightness ratio 296bp band brightness is low, and P value is respectively less than 0.5, therefore, it is determined that being beard gene heterozygous.
3 test individual PCR product band luminance quantization value of table and genotype determine
| Sample number | 296bp band brightness | 207bp band brightness | P value | Genotype |
| 1 | 135.2 | 118.9 | 0.88 | Beard gene is homozygous |
| 2 | 142.7 | 116.3 | 0.82 | Beard gene is homozygous |
| 3 | 159.1 | 126.8 | 0.80 | Beard gene is homozygous |
| 4 | 284.3 | ? | 0 | Non- beard gene |
| 5 | 167.8 | 69.1 | 0.41 | Beard gene heterozygous |
| 6 | 365.5 | ? | 0 | Non- beard gene |
| 7 | 166.3 | 60.0 | 0.36 | Beard gene heterozygous |
| 8 | 137.7 | 58.8 | 0.43 | Beard gene heterozygous |
| 9 | 183.5 | 74.9 | 0.41 | Beard gene heterozygous |
| 10 | 363.9 | ? | 0 | Non- beard gene |
| 11 | 114.2 | 45.6 | 0.40 | Beard gene heterozygous |
| 12 | 127.3 | 49.1 | 0.39 | Beard gene heterozygous |
| 13 | 297.0 | ? | 0 | Non- beard gene |
| 14 | 228.9 | ? | 0 | Non- beard gene |
| 15 | 149.3 | 106.7 | 0.71 | Beard gene is homozygous |
| 16 | 146.3 | 106.3 | 0.73 | Beard gene is homozygous |
| 17 | 205.4 | 143.8 | 0.70 | Beard gene is homozygous |
| 18 | 222.4 | ? | 0 | Non- beard gene |
| 19 | 182.2 | 140.2 | 0.77 | Beard gene is homozygous |
| 20 | 68.1 | 24.7 | 0.36 | Beard gene heterozygous |
As a result as it can be seen that the naked eyes of band brightness set it is completely the same with the genotype of software identification decision beard gene, with
The actual gene type that conceals it was found that, determination rate of accuracy is up to 100%.
The present invention carries out the common multi-PRC reaction of single merely with 3 primers, can successfully identify the pure of beard chicken
Zygote, heterozygote and non-beard chicken, method is simple, fast and accurately, the very useful breeding in beard chicken spiral trough and
Purification.Specific advantage is as follows:
(1) the genotype molecule identification of chicken beard character gene of the present invention, can identify pure under phenotype same case
Mould assembly and heterozygous individual, effectively increase the accuracy of seed selection；
(2) Molecular Identification of the present invention is not limited by age, gender of beard chicken etc., can be used for beard character
The generation inteval is shortened in early stage breeding, accelerates beard a breed of chicken process；
(3) detection method is simple, quick, accurate.
The sensitivity technique of 4 primer of embodiment is tested
Non- beard chicken, beard chicken giblets zygote and beard chicken homozygote each 1 are chosen respectively, genomic DNA are extracted, by it
DNA concentration is diluted to 100 μ g/mL, 50 μ g/mL, 20 μ g/mL and 10 μ g/mL respectively,
The amplification program of PCR reaction and the method for amplified production detection are the same as embodiment 2 and 3, PCR amplification 27 circulations.Expand
Increase after reaction, electrophoresis detection is carried out to amplified production with 2% Ago-Gel, as a result as shown in Figure 7.1-4 swimming lane point
DNA concentration in PCR reaction is not represented and is followed successively by 100 μ g/mL, 50 μ g/mL, 20 μ g/mL and 10 μ g/mL, it can be seen that each gene
Bright-dark degree in type on 1 and 2 swimming lanes between product band is the most obvious, and While it can be seen that PCR is produced on 3 and 4 swimming lanes
Object band, but band brightness is inadequate, is not suitable for distinguishing PCR product amount by bright-dark degree, to the homozygous and miscellaneous of beard gene
The band of mould assembly carrys out large error.
From the point of view of Fig. 7, although DNA concentration range can amplify characteristic bands, and 3 primers in 20-100 μ g/mL
Minimum can be fixed on 20 μ g/mL by DNA template concentration wider range needed for PCR amplification.But combine the present invention maximum
Characteristic identifies the homozygous and heterozygous of beard chicken by the bright-dark degree of interband, is also convenient for ImageJ software brightness value
Calculating, therefore template concentrations for finally reacting the DNA concentration of 50-100 μ g/mL as PCR of the present invention.
Although above the present invention is described in detail with a general description of the specific embodiments,
On the basis of the present invention, it can be made some modifications or improvements, this will be apparent to those skilled in the art.Cause
This, these modifications or improvements, fall within the scope of the claimed invention without departing from theon the basis of the spirit of the present invention.
Claims (9)
1. the PCR primer combination for identifying chicken beard character gene type characterized by comprising
Forward primer F1:5'-TGGTTGTGTGCCAAGTAGAG-3'；
Forward primer F2:5'-GCTCTTGCTCTTTTTGGTAA-3'；And
Reverse primer R:5'-CTTTTCGTACTGCGTTGTTA-3'.
2. containing the combination of primer described in claim 1 for identifying the kit of chicken beard character gene type.
3. the gene type rapid identification method of chicken beard character, which comprises the following steps:
1) genomic DNA of chicken to be measured is extracted；
2) it using the DNA extracted in step 1) as template, is combined using primer described in claim 1, carries out pcr amplification reaction；
3) PCR product is analyzed.
4. according to the method described in claim 3, it is characterized in that, PCR reaction system is calculated as with 20 μ l:
5. according to the method described in claim 3, it is characterized in that, PCR reaction condition are as follows: 95 DEG C 5 minutes；95 DEG C 30 seconds, 53
DEG C 30 seconds, 72 DEG C 20 seconds, totally 27 circulations；72 DEG C 7 minutes.
6. according to the described in any item methods of claim 3-5, which is characterized in that the method that step 3) analyzes PCR product is as follows:
Pcr amplification product is subjected to 2% agarose gel electrophoresis detection, ethidium bromide staining observes PCR in gel imaging system
Amplified band, and take pictures；
With Image J software analytical electrophoresis glue figure, the brightness value of 296bp band and 207bp band in each swimming lane is calculated, and count
Calculate the ratio P of 207bp band brightness value and 296bp band brightness value；P value is determined as that beard chicken gene is pure between 0.7-0.9
Zygote, P value are determined as beard chicken giblets zygote between 0.2-0.5；P value is 0, only amplifies 296bp band, is then determined as non-Hu
It must chicken individuals.
7. specific molecular marker relevant to chicken beard character, which is characterized in that the molecular labeling is to be located at chicken No. 27 dyes
The downstream CNV1 on colour solid, size is the DNA fragmentation of 207bp, for expanding the primer of the molecular labeling are as follows:
Forward primer F1:5'-TGGTTGTGTGCCAAGTAGAG-3' and
Reverse primer R:5'-CTTTTCGTACTGCGTTGTTA-3'.
8. application of the specific molecular marker described in claim 7 in identification beard chicken breed.
9. application of the specific molecular marker described in claim 7 in chicken molecular mark.
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| CN115131784B (en) * | 2022-04-26 | 2023-04-18 | 东莞博奥木华基因科技有限公司 | Image processing method and device, electronic equipment and storage medium |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104342489A (en) * | 2013-08-02 | 2015-02-11 | 中国农业大学 | Method for detecting genotype of chicken beard |
| CN104342490A (en) * | 2013-08-02 | 2015-02-11 | 中国农业大学 | Method for detecting chicken beard traits |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104342489A (en) * | 2013-08-02 | 2015-02-11 | 中国农业大学 | Method for detecting genotype of chicken beard |
| CN104342490A (en) * | 2013-08-02 | 2015-02-11 | 中国农业大学 | Method for detecting chicken beard traits |
Non-Patent Citations (2)
| Title |
|---|
| A Complex Structural Variation on Chromosome 27 Leads to the Ectopic Expression of HOXB8 and the Muffs and Beard Phenotype in Chickens;Ying Guo等;《PLOS GENETICS》;20160602;第12卷(第6期);e1006071 |
| 鸡胡须形状的基因定位及功能研究;郭影;《中国博士学位论文全文数据库 农业科技辑》;20160815(第08期);D050-92 |
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