CN116287287B - Molecular detection method applied to small Bai Ji green foot character and application thereof - Google Patents
Molecular detection method applied to small Bai Ji green foot character and application thereof Download PDFInfo
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Abstract
The invention discloses a molecular detection method applied to the property of small Bai Ji green feet and application thereof, belonging to the technical field of biological molecules. The molecular detection method comprises the following steps: taking the genomic DNA of the chicken to be detected as a template, and carrying out PCR amplification on the gene sequence fragment of the SEQ ID NO.6 of the white chicken by using a primer pair Q to obtain an amplified product; and digesting the amplification product by using restriction endonuclease, performing agarose gel electrophoresis on the amplified fragment after enzyme digestion, and identifying SNP locus genotype of CM028522.1:g.26049541 according to electrophoresis results, thereby detecting the green foot character dominant haplotype type of the white chicken. The invention also provides application of the primer pair Q in genetic breeding of chickens. The method can be used for molecular marker assisted selection of the small Bai Ji green foot character, accelerates the shank breeding process of the white chickens and meets different market demands.
Description
Technical Field
The invention relates to the technical field of biological molecules, in particular to a molecular detection method applied to the characteristic of small Bai Ji green feet and application thereof.
Background
The chicken green foot character is a character with important economic significance, and the purchasing desire of consumers is closely related to the color of the chicken feet of the carcasses, so that the shank color of the carcasses directly influences the sales of the broiler market. Studies have shown that the green foot trait is controlled primarily by dermal melanin inhibitors (dermal melanin inhibitor, id) on the Z chromosome, with the shin appearing as a light shin if the individual has an Id gene, melanin is not deposited on the shin; if an individual only has an id gene, the shin is melanin deposited and the shin color appears as a dark shin. Therefore, offspring with specific shank colors can be bred by utilizing the characteristic of the green foot character concomitant inheritance. Major genes or key causative mutations for regulating green foot traits have not been precisely located so far.
The white chickens are hybrid matched varieties of broiler chickens, the problems of poor uniformity of the shank color and difficult fixation of the shank color of commercial generation hens are common in the current production practice of the white chickens, and the proportion of green feet to yellow feet in hen groups is about 1:1. In the large-scale production of the white chickens, the traditional breeding method cannot fix the shin color of the white chickens, the breeding process of the shin color is slow, and a molecular detection method applied to the green foot character of the small Bai Ji is urgently needed.
At present, the molecular markers related to the shank color of the broiler chickens are relatively few, and particularly, a shank color identification method aiming at a white chicken complete set is not reported. The Dorshorst localization Id in 2010 was within the 67.1-72.3Mb (galgal 3.0) interval associated with the shank trait for the Z chromosome; SNPs of GRAMD3, which were significantly associated with the dermal pigment repressor gene, were found in fixed-born chickens in 2017 in the 79.02-79.44Mb (galGal 4) interval; in 2022, SNPs of MC1R and CDKN2A related to 817-class meat-mixed chicken green foot traits are found, but the molecular markers are not suitable for accurate phenotype identification of the shank color of commercial chicken generation of white chickens, have no linkage relationship with the small Bai Ji green foot traits, and cannot be applied to molecular breeding related to the small Bai Ji green foot traits. Therefore, if an accurate detection method applicable to the characteristics of the green feet of the small Bai Ji can be provided, the breeding process of the white chickens is greatly accelerated, the problem of poor carcass uniformity of the white chickens is overcome, the white chickens are promoted to slaughter and market transformation and upgrading, and the sales profits of the white chickens are remarkably improved.
Disclosure of Invention
The invention aims to provide a molecular detection method applied to the characteristic of the green foot of Bai Ji and application thereof, so as to solve the problems in the prior art, detect SNP loci obviously related to the characteristic of the green foot by using the molecular detection method, and select haplotype individuals according to production requirements, thereby accelerating the establishment of white chicken varieties with specific shank colors.
In order to achieve the above object, the present invention provides the following solutions:
the technical scheme is as follows: a molecular detection method for detecting dominant haplotype type of green foot characters of white chickens comprises the following steps:
(1) Taking the genomic DNA of the chicken to be detected as a template, and carrying out PCR amplification on the gene sequence fragment of the SEQ ID NO.6 of the white chicken by using a primer pair Q to obtain an amplified product;
(2) And (3) digesting the PCR amplification product by using restriction endonuclease, performing agarose gel electrophoresis on the amplified fragment after enzyme digestion, and identifying the SNP locus genotype of CM028522.1:g.26049541 according to the electrophoresis result, thereby detecting the green foot character dominant haplotype type of the white chicken.
Further, in step (1), the primer pair Q includes an upstream primer of the nucleotide sequence shown in SEQ ID NO.3 and a downstream primer of the nucleotide sequence shown in SEQ ID NO. 4.
Further, in step (1), the PCR amplification system is: 2 XTaq polymerase premix 5. Mu.L, upstream primer 1. Mu.L, downstream primer 1. Mu.L, template DNA 1. Mu.L and double distilled water 2. Mu.L; the PCR amplification reaction program is as follows: pre-denaturation at 95℃for 2min; denaturation at 95℃for 15s, annealing at 60℃for 15s, elongation at 72℃for 17s,30 cycles; thoroughly extending at 72 ℃ for 5min; preserving at 4 ℃.
Further, in step (2), the haplotype consists of the SNP1-SNP21 sites within the 25.96Mb-26.05Mb region of the Z chromosome.
Further, the SNP1-SNP21 is located in CM028522.1, wherein SNP1 is located in g.25959465, and the mutation site is C > T; SNP2 is positioned in g.25964271, and the mutation site is A > C; SNP3 is positioned in g.25965995, and the mutation site is A > G; SNP4 is positioned in g.25966438, and the mutation site is G > C; SNP5 is positioned in g.25985562, and the mutation site is A > G; SNP6 is positioned in g.25989827, and the mutation site is G > A; SNP7 is positioned in g.25991831, and the mutation site is C > T; SNP8 is positioned in g.25994954, and the mutation site is T > C; SNP9 is positioned in g.25994972, and the mutation site is A > G; the SNP10 is positioned in g.25999838, and the mutation site is C > G; SNP11 is positioned in g.26016056, and the mutation site is T > G; SNP12 is positioned in g.26043162, and the mutation site is A > C; SNP13 is positioned in g.26047179, and the mutation site is C > A; SNP14 is positioned in g.26047223, and the mutation site is G > C; SNP15 is positioned in g.26049519, and the mutation site is T > C; SNP16 is positioned in g.26049520, and the mutation site is G > A; SNP17 is positioned in g.26049531, and the mutation site is A > T; SNP18 is positioned in g.26049541, and the mutation site is G > C; SNP19 is positioned in g.26049556, and the mutation site is C > T; the SNP20 is positioned in g.26049736, and the mutation site is G > A; SNP21 is positioned in g.26049759, and the mutation site is C > T.
Further, in the step (2), the restriction enzyme is Bfo I, and the restriction enzyme cleavage system is: 10. Mu.L of amplification product, 0.7. Mu.L of 10 Xfast-digestion enzyme buffer, and 0.3. Mu.L of fast-digestion enzyme Bfo I; the reaction procedure of the enzyme digestion is as follows: digesting for 90min at 95 ℃; digestion was stopped at 65℃for 5 min.
Further, in the step (2), the identification of the SNP locus genotype of CM028522.1:g.26049541 based on the result of electrophoresis comprises: when the electrophoresis result shows a 500bp (SEQ ID NO. 6) band, the band is a genotype 1 cock or hen, and the cock genotype is Z G Z G Hen genotype Z G W is a metal; when the electrophoresis result shows that the bands are two and the sizes are 273bp (SEQ ID NO. 7) and 227bp (SEQ ID NO. 8), the bands are genotype 2 cock or hen, and the cock genotype is Z C Z C Hen genotype Z C W is a metal; when the electrophoresis result shows that the bands are three and the sizes are 500bp,273bp and 227bp respectively, the bands are heterozygous cock, and the cock genotype is Z C Z G 。
Further, in the step (2), the agarose gel electrophoresis is determined as follows: the digested PCR product was aspirated at 10. Mu.L, and the sample was spotted into a 3% agarose TAE gel and electrophoresed for 20min, and the result of nucleic acid electrophoresis was observed under an ultraviolet lamp.
The second technical scheme is as follows: an application of the primer pair Q in genetic breeding of chickens.
Further, the genetic breeding of chickens comprises breeding green-foot white chickens or reducing the green-foot ratio of white chickens.
The invention discloses the following technical effects:
according to the invention, 11,141,498 SNP loci with high credibility are finally screened out by a whole gene resequencing technology and a GWAS analysis method. The high-quality SNP loci and the shank color phenotype are subjected to genome-wide association analysis by using two analysis methods of chi-square test (- -assoc) and Fisher exact test (- -Fisher), and 22 significant loci are co-located, wherein 21 common significant loci are all located in a 25.96Mb-26.05Mb interval on a Z chromosome. The 21 SNP loci are completely linked and can be divided into haplotype 1 and haplotype 2. The invention selects one SNP18 of the obvious loci, and identifies the genotype of the sample by an RFLP-PCR method, thereby determining the haplotype condition of the individual to be detected. The method does not need DNA sequence determination, has high accuracy, simple operation and detection cost saving, is suitable for detecting the green foot character of the small Bai Ji, and has important significance for shank color breeding of white chicken varieties.
Because 21 obvious SNP loci are closely related to green foot traits in a 25.96Mb-26.05Mb region of a chicken Z chromosome, the loci are completely linked and can be divided into haplotype 1 and haplotype 2. The genotype of SNP18 of one of the obvious loci is identified by a PCR-RFLP method, and the dominant haplotype type of the small Bai Ji green foot trait can be detected. The method has the advantages of no need of DNA sequence determination, high accuracy, simple operation, detection cost saving and suitability for detecting the small Bai Ji green foot characters. The invention selects one SNP18 of the obvious loci, and identifies the genotype of the sample by an RFLP-PCR method, thereby identifying the dominant haplotype type of the green foot character of the white chicken. In the production of raising poultry, the variety of green-foot chickens can be selected from genotype 1 individuals according to breeding requirements, or the green-foot proportion of the white chickens can be reduced by eliminating genotype 1 individuals.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a Manhattan diagram of a chi-square test (- -assoc);
FIG. 2 is a Fisher exact test (- -Fisher) Manhattan diagram;
FIG. 3 is a diagram showing SNP typing and peak of hens;
FIG. 4 is a graph of cock SNP typing versus peak;
FIG. 5 is a graph showing the effect of molecular detection on genotyping of chicken by RFLP-PCR, wherein lanes 1,2, and 3: z is Z G Z G Genotype cock (500 bp); lanes 4,5,6: z is Z C Z G Genotype cock (500 bp,273bp and 227 bp); lanes 7,8,9: z is Z C Z C Genotype cock (273 bp and 227 bp); lanes 10, 11, 12: z is Z G W genotype hens (500 bp); lanes 13, 14, 15: z is Z C W genotype hens (273 bp and 227 bp); m: marker (500 bp,400bp,300bp,250bp,200bp,150bp,100bp,50 bp).
Detailed Description
Various exemplary embodiments of the invention will now be described in detail, with reference to the examples using conventional methods, unless otherwise indicated, and with reference to reagents, either conventional commercial reagents or reagents configured using conventional methods. The detailed description is not to be taken as limiting, but is to be understood as a more detailed description of certain aspects, features, and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. In addition, for numerical ranges in this disclosure, it is understood that each intermediate value between the upper and lower limits of the ranges is also specifically disclosed. Every smaller range between any stated value or stated range, and any other stated value or intermediate value within the stated range, is also encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the invention described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from consideration of the specification of the present invention. The specification and examples of the present invention are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are intended to be inclusive and mean an inclusion, but not limited to.
In the following description, unless otherwise indicated, all methods involved are conventional in the art. Unless otherwise indicated, all materials referred to are those available from published commercial sources.
Example 1
1 materials and methods
1.1 animal sample
Using 101 cobra cocks (female) and 404 yellow-feather broiler hens (female) 1:4, crossing to establish a resource family. 196F 1 hens and 72 male parents were selected, and 2mL of subcutaneous venous blood was collected and stored at-80℃for use as DNA extraction samples.
1.2 major reagents
Blood sample DNA extraction kit (brand: OMEGA; cat# D3392; guangzhou Hi-Bo Biotechnology Co., ltd.), 2 Xfast Taq polymerase premix (brand: novepran; cat# P222-01; jiangsu Norvezan Biotechnology Co., ltd.), DNAmaroker (brand: gold sand; cat# SM815-500; beijing gold sand Biotechnology Co., ltd.), high purity low electroosmotic agarose (brand: opt-J001; beijing Biotechnology Co., ltd.), fast digestion restriction endonuclease Bfo I (brand: simer F.; cat# FD2184; simer Fei.Tech., ltd.).
2 Experimental methods
2.1 blood genomic DNA extraction
Blood sample DNA was extracted by reference to the animal blood sample DNA extraction kit operating manual.
2.2 Whole genome resequencing and mutation detection
Representative yellow feet (39) and green feet (40) individuals are collected, genomic DNA is extracted, and SNP loci with high reliability are finally screened out through a whole gene resequencing technology and a GWAS analysis method. And carrying out genome-wide association analysis on the high-quality SNP locus and the shank phenotype by using two analysis methods of chi-square test (- -assoc) and Fisher accurate test (- -Fisher), and positioning the SNP locus with obvious association.
2.3 resequencing results analysis and validation
The common significant locus genotype of 80 re-sequenced individuals was analyzed and haplotypes were classified for the loci. Analysis of sequence peak patterns was performed on Sanger sequencing results of PCR products using SeqMan tool of DNAstar software, 117 half-sibling individuals except for resequencing were genotyped, and the duty ratios of the genotypes in the various shank colors were counted.
2.4 primer design
The red chicken (Gallus) Z chromosome genomic sequence published under NCBI (National Center for Biotechnology Information Search database) (ID: CM 028522.1). Primers were designed using NCBI's Primer-BLAST tool, offered by the Guangzhou Prime Biotechnology Co., ltd. The primer sequences are shown in Table 1.
TABLE 1 PCR amplification primer sequences
2.5 PCR amplification of target sequences
A PCR amplification reaction system was prepared using chicken blood genomic DNA as a template (Table 2). The PCR reaction system is shown in Table 3. The PCR products were sequenced and Sanger sequencing was performed by Sanger, inc. of Tian Yihui Yuan Gene technology, guangzhou.
TABLE 2 reaction system
Table 3 reaction procedure
2.6RFLP-PCR method for detecting genotype
The RFLP-PCR primer is primer pair Q, and the nucleotide sequence is shown as SEQ ID NO.3 and SEQ ID NO. 4. The RFLP-PCR reaction system and the procedure are shown in tables 4 and 5. The digested products were genotyped using agarose gel electrophoresis.
TABLE 4 reaction system
Table 5 reaction procedure
3 results
3.1 genome-wide resequencing and mutation detection positioning control of Gene locus of white chicken shin
Through the whole gene resequencing technology and the GWAS analysis method, 11,141,498 SNP loci with high reliability are finally screened out. The high quality SNP sites were subjected to genome-wide association analysis with the tibial phenotype using two analysis methods, chi-square test (Chi-SquareTest) and Fisher's exact test (0.05/snprint) as thresholds, to locate 22 (FIG. 1) and 41 (FIG. 2) significantly associated sites, respectively. Intersection analysis of the results obtained from the two test methods revealed 22 common significant loci (Table 6), 21 of the 22 significant loci were located in the 25.96Mb-26.05Mb interval on the Z chromosome, confirming that the small Bai Ji green foot trait is a companion genetic trait controlled by the Z chromosome.
TABLE 6 chi-square test and Fisher's exact test co-detected significant sites
3.2 resequencing results genotyping case
Analysis of 21 significant sites on the Z chromosome revealed that the above sites were completely linked and were divided into two haplotypes, haplotype 1 and haplotype 2 (Table 7). Analysis of the correlation between the two haplotypes and the shank of the F1 hen revealed that the frequency of occurrence of haplotype 1 in the green-foot hen was 92.73%, that of occurrence of haplotype 2 was only 7.27%, that of occurrence of haplotype 1 in the yellow-foot hen was 43.48%, that of haplotype 2 was 56.52% (Table 8), and that haplotype 1 was the dominant haplotype regulating the green-foot trait.
TABLE 7 haplotype composition of Z chromosomal significant loci
TABLE 8 haplotype distribution of different shank hens
Verification of genome-wide association analysis results of 3.3F1 offspring population
In view of the complete linkage of 21 significant loci on the Z chromosome, 7 SNP loci are randomly selected to genotype and genotype the 72 test cross male parents and the rest F1 hens. The above-mentioned SNP is SNP8, SNP9, SNP15, SNP16, SNP17, SNP18, SNP19.SNP8 and SNP9 are located at 251bp and 269bp of SEQ ID NO. 5, respectively. SNP15 to SNP19 are located at 251bp, 252bp, 263bp, 273bp and 288bp of SEQ ID NO.6, respectively. 117F 1 hens, except for resequencing, were selected and PCR amplified using primer pair a and primer pair Q, with the DNA of the blood sample of each individual as a template. Peak patterns after Sanger sequencing of PCR products were analyzed by comparison. The hen sequencing peak pattern of 7 SNP sites is shown in FIG. 3, and the cock sequencing peak pattern is shown in FIG. 4.
SEQ ID NO:5：
CATTTGTCCTGATATTTATCTGCCTGCCAGGTTAAACCACAACACCACTATAAATGAATATTGCCAATATCACTAATCAAAATACTCCTTGACATAAATCATTACCAAAAAAATCAGGTATGCTGAAGCAGTAATAGGGAACAAATAAGAGTGAAGAAAATCAATACGTCAACACTGAAGCAGGACCCGCAACTTAGACACTTTCATAAAAAGGAGCTTCATCTTGGACACAGAAGAGCCTCCTTACACATTGTATGATACTCTACAGAGTCAAGATAAACTGCAAGAAGGGCTTTGAAGTAGTGTACTTTTTTGCACTTATCACCTGAAAGATCTGTATTAGTCATACATCCACCACGATCCTATCTTAAAAAAGGGAAGCAACGTCCACATCTGATAGAGTCAGACAATTTATCCCCTTTCTCTAAGCATCACATGGGGGAGGGAAGGGGAGAGAAGTTATTGATTTTGTTTCTCTTGTAGACAATTGCAATGACTGGTAGCATAACGTGAGCAAGA。
Genotype analysis of the test cross male parent revealed that 7 significant loci of all male parents were completely linked and divided into 3 genotypes, genotype 1 to genotype 3 respectively, wherein genotype 3 was heterozygous for genotype 1 and genotype 2 (table 9). In the male parent genotype, the proportion of genotype 1 is up to 62.5%, which is the dominant genotype.
TABLE 9 genotyping results for 7 significant loci of male parent
Genotype analysis was performed on 7 significant sites of 117F 1 hens except for resequencing, and the results showed that 7 SNP sites were completely linked, and the genotype composition was consistent with resequencing results and classified into haplotype 1 and haplotype 2 (Table 10). The result of the combined analysis of the genotype of the F1 hen and the shank color identification of the 43-day-old carcasses shows that the green foot proportion of the haplotype 1 hen is 80.82 percent, while the green foot proportion of the haplotype 2 hen is 54.00 percent (table 11), which indicates that the rooster with the genotype 1 is removed, and the rooster with the genotype 2 is reserved, so that the green foot proportion can be effectively reduced.
TABLE 10 7 significant site haplotype composition for 117 hens except for resequencing
TABLE 11 statistics of all F1 hen genotypes and shank color results
3.4RFLP-PCR method for detecting genotype of white chicken
Since 21 significant sites are completely linked, the genotype of only one SNP needs to be identified to map the individual haplotype situation. SNP18 is selected as a SNP genotype detection site, and the genotype is taken as Z G Z G 、Z G Z C 、Z C Z C Is Z in the cock and genotype G W、Z C W hen whole genome DNA is used as a template, and a primer pair Q is used for amplifying a gene sequence fragment of the white chicken SEQ ID NO. 6. And (3) digesting the PCR amplified product by using restriction endonuclease Bfo I, then performing agarose gel electrophoresis on the amplified fragment after enzyme digestion, and detecting SNP18 genotype according to the electrophoresis result. In the agarose gel electrophoresis typing chart (FIG. 5), lanes 1,2,3 are Z G Z G Genotype cock with only one 500bpA strap. Lanes 4,5,6 are Z C Z G The genotype cock has bands of 500bp (SEQ ID NO. 6), 273bp (SEQ ID NO. 7) and 227bp (SEQ ID NO. 8), respectively. Lanes 7,8,9 are Z C Z C The genotype cock has two 273bp and 227bp bands generated after incision by endonuclease. Similarly lanes 10, 11, 12 are Z G W-genotype hen, electrophoresis strip and Z G Z G The genotype cock is the same. Lanes 13, 14, 15 are Z C W-genotype hen, electrophoresis gel pattern exhibited Z C Z C Genotypic rooster-like bands.
SEQ ID NO.6：
ATGCAGGAAAGCAGAACACATACCCCAAATCACAGTTCTGCCATGGAATTAGCACCAGGTACCTCGTGGGATGCAGAAGAGGTGAAGAAGAAAACTGAATCAAAATGGGAGCTTAAATGAAGCATATGTCTTGTGGTCTTTATTCAGTTGTTTATGGGAACACGGTTTCAGGTCTACACTTTCCTAGCACGTAATGCTAAAGTTATGATAGCTACTTCATTGTTAATCAACTTCTGAAGGAAGTGAGGCTTGAGTGCAGATCACCCAAGGCGGCTAAATTCATGCACACAAAGGTACCCAGCGCAAGTATACATTTTATTGACTCTTTAGGTTTTTTACTCCAGCTTCCCCCCAAGTTTGATTATCCAGATTTGCCTATTTCTTCA GTACTGCTAAAAATCAAAGTTGTGTCATGAACTCTTTTCATCATTAGAATATTATGGGGTGTACAATCTGGTGCTTGAAGAGGAAAAGCTGTTCTTTGTGTGGCACATAGTTACA。
SEQ ID NO.7：
ATGCAGGAAAGCAGAACACATACCCCAAATCACAGTTCTGCCATGGAATTAGCACCAGGTACCTCGTGGGATGCAGAAGAGGTGAAGAAGAAAACTGAATCAAAATGGGAGCTTAAATGAAGCATATGTCTTGTGGTCTTTATTCAGTTGTTTATGGGAACACGGTTTCAGGTCTACACTTTCCTAGCACGTAATGCTAAAGTTATGATAGCTACTTCATTGTTAATCAACTTCTGAAGGAAGTGAGGCTTGAGTGCAGATCACCCAAGGCGG。
SEQ ID NO.8：
TAAATTCATGCACACAAAGGTACCCAGCGCAAGTATACATTTTATTGACTCTTTAGGTTTTTTACTCCAGCTTCCCCCCAAGTTTGATTATCCAGATTTGCCTATTTCTTCAGTACTGCTAAAAATCAAAGTTGTGTCATGAACTCTTTTCATCATTAGAATATTATGGGGTGTACAATCTGGTGCTTGAAGAGGAAAAGCTGTTCTTTGTGTGGCACATAGTTACA。
The result shows that the haplotype condition of the chicken can be accurately distinguished by the method. Therefore, the SNP18 genotype of the white chicken is detected by utilizing the RFLP-PCR method, and the haplotype condition of the white chicken can be further deduced. The RFLP-PCR method is a molecular method related to the characteristic of small Bai Ji green feet, and can be used for detecting haplotype molecules in breeding white chickens. The single-type 1 individuals are removed, so that the proportion of green feet of the population can be effectively reduced, the single-type 1 individuals are reserved, and the breeding process of green foot white chicken varieties can be accelerated.
The above embodiments are only illustrative of the preferred embodiments of the present invention and are not intended to limit the scope of the present invention, and various modifications and improvements made by those skilled in the art to the technical solutions of the present invention should fall within the protection scope defined by the claims of the present invention without departing from the design spirit of the present invention.
Claims (5)
1. The molecular detection method for detecting the dominant haplotype type of the green foot character of the white chicken is characterized by comprising the following steps of:
(1) Taking the genomic DNA of the chicken to be detected as a template, and carrying out PCR amplification on the SEQ ID NO.6 gene sequence fragment of the white chicken by using a primer pair Q to obtain an amplified product;
(2) Digesting the amplification product by using restriction endonuclease, performing agarose gel electrophoresis on the amplified fragment after enzyme digestion, and identifying SNP locus genotype of CM028522.1:g.26049541 according to electrophoresis result, thereby detecting green foot character dominant haplotype type of white chickens;
in step (1), the primer pair Q comprises an upstream primer having a nucleotide sequence shown as SEQ ID NO.3 and a downstream primer having a nucleotide sequence shown as SEQ ID NO. 4;
in step (2), the haplotype consists of the SNP1-SNP21 sites within the 25.96Mb-26.05Mb region of the Z chromosome;
the SNP1-SNP21 is positioned in CM028522.1, wherein SNP1 is positioned in g.25959465, and the mutation site is C > T; SNP2 is positioned in g.25964271, and the mutation site is A > C; SNP3 is positioned in g.25965995, and the mutation site is A > G; SNP4 is positioned in g.25966438, and the mutation site is G > C; SNP5 is positioned in g.25985562, and the mutation site is A > G; SNP6 is positioned in g.25989827, and the mutation site is G > A; SNP7 is positioned in g.25991831, and the mutation site is C > T; SNP8 is positioned in g.25994954, and the mutation site is T > C; SNP9 is positioned in g.25994972, and the mutation site is A > G; SNP10 is positioned in g.25999838, and the mutation site is C > G; SNP11 is positioned in g.26016056, and the mutation site is T > G; SNP12 is positioned in g.26043162, and the mutation site is A > C; SNP13 is positioned in g.26047179, and the mutation site is C > A; SNP14 is positioned in g.26047223, and the mutation site is G > C; SNP15 is positioned in g.26049519, and the mutation site is T > C; SNP16 is positioned in g.26049520, and the mutation site is G > A; SNP17 is positioned in g.26049531, and the mutation site is A > T; SNP18 is positioned in g.26049541, and the mutation site is G > C; SNP19 is positioned in g.26049556, and the mutation site is C > T; the SNP20 is positioned in g.26049736, and the mutation site is G > A; SNP21 is positioned in g.26049759, and the mutation site is C > T;
in the step (2), the identification of the SNP locus genotype of CM028522.1:g.26049541 according to the electrophoresis result comprises: when the electrophoresis result shows a 500bp band, the band is a genotype 1 cock or hen, and the cock genotype is Z G Z G Hen genotype Z G W is a metal; when the electrophoresis result shows that the bands are two and the sizes are 273bp and 227bp respectively, the bands are genotype 2 cock or hen, and the genotype of the cock is Z C Z C Hen genotype Z C W is a metal; when the electrophoresis result shows that the bands are three and the sizes are 500bp,273bp and 227bp respectively, the bands are heterozygous cock, and the cock genotype is Z C Z G 。
2. The method of claim 1, wherein in step (1), the PCR amplification system is: 2 XTaq polymerase premix 5. Mu.L, upstream primer 1. Mu.L, downstream primer 1. Mu.L, template DNA 1. Mu.L and double distilled water 2. Mu.L; the PCR amplification reaction program is as follows: pre-denaturation at 95℃for 2min; denaturation at 95℃for 15s, annealing at 60℃for 15s, elongation at 72℃for 17s,30 cycles; thoroughly extending at 72 ℃ for 5min; preserving at 4 ℃.
3. The molecular detection method according to claim 1, wherein in the step (2), the restriction enzyme is Bfo I, and the cleavage system of the restriction enzyme is: 10. Mu.L of amplification product, 0.7. Mu.L of 10 Xfast-digestion enzyme buffer, 0.3. Mu.L of fast-digestion enzyme Bfo I; the reaction procedure of the enzyme digestion is as follows: digesting for 90min at 95 ℃; digestion was stopped at 65℃for 5 min.
4. The method for detecting a molecule according to claim 1, wherein in the step (2), the agarose gel electrophoresis is determined by: the digested PCR product was aspirated at 10. Mu.L, and the sample was spotted into a 3% agarose TAE gel and electrophoresed for 20min, and the result of nucleic acid electrophoresis was observed under an ultraviolet lamp.
5. The application of the primer pair Q in the genetic breeding of chickens is characterized in that the genetic breeding of chickens comprises the steps of breeding green-foot white chicken varieties or reducing the green-foot ratio of white chicken groups; the primer pair Q comprises an upstream primer with a nucleotide sequence shown as SEQ ID NO.3 and a downstream primer with a nucleotide sequence shown as SEQ ID NO. 4.
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Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104293905A (en) * | 2014-04-11 | 2015-01-21 | 河南农业大学 | Primer, kit and detection method for detecting chicken green shin character linkage SNP locus genotype |
| KR20150050499A (en) * | 2013-10-30 | 2015-05-08 | 충남대학교산학협력단 | A method for discriminating of shank color of korean native chicken |
| CN104719253A (en) * | 2015-04-15 | 2015-06-24 | 广西南宁市富凤农牧有限公司 | Breeding method of low-speed type super-quality green-leg parental breeding chickens |
| WO2019071407A1 (en) * | 2017-10-10 | 2019-04-18 | 中国农业科学院北京畜牧兽医研究所 | Snp chip for whole chicken genome and application thereof |
| WO2019128350A1 (en) * | 2017-12-29 | 2019-07-04 | 北京市华都峪口禽业有限责任公司 | Identification of white leghorns red plumage mutagenic mutant genotype and cultivation method for supporting system of red plumage pink shell layer chickens |
| CN110578008A (en) * | 2019-10-28 | 2019-12-17 | 东北农业大学 | molecular marking method for predicting and identifying chicken abdominal fat weight |
| CN112662790A (en) * | 2021-01-29 | 2021-04-16 | 南昌师范学院 | SNP molecular marker related to green-foot character of 817 meat hybrid chickens and application thereof |
| CN112746113A (en) * | 2021-01-29 | 2021-05-04 | 南昌师范学院 | Molecular marker related to chicken green foot character and application thereof |
| CN114214432A (en) * | 2021-12-31 | 2022-03-22 | 江苏北农大农牧科技有限公司 | Molecular marker for identifying melanin diffusion gene locus of chicken and application of molecular marker in assisted breeding |
-
2022
- 2022-12-13 CN CN202211625745.8A patent/CN116287287B/en active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20150050499A (en) * | 2013-10-30 | 2015-05-08 | 충남대학교산학협력단 | A method for discriminating of shank color of korean native chicken |
| CN104293905A (en) * | 2014-04-11 | 2015-01-21 | 河南农业大学 | Primer, kit and detection method for detecting chicken green shin character linkage SNP locus genotype |
| CN104719253A (en) * | 2015-04-15 | 2015-06-24 | 广西南宁市富凤农牧有限公司 | Breeding method of low-speed type super-quality green-leg parental breeding chickens |
| WO2019071407A1 (en) * | 2017-10-10 | 2019-04-18 | 中国农业科学院北京畜牧兽医研究所 | Snp chip for whole chicken genome and application thereof |
| WO2019128350A1 (en) * | 2017-12-29 | 2019-07-04 | 北京市华都峪口禽业有限责任公司 | Identification of white leghorns red plumage mutagenic mutant genotype and cultivation method for supporting system of red plumage pink shell layer chickens |
| CN110578008A (en) * | 2019-10-28 | 2019-12-17 | 东北农业大学 | molecular marking method for predicting and identifying chicken abdominal fat weight |
| CN112662790A (en) * | 2021-01-29 | 2021-04-16 | 南昌师范学院 | SNP molecular marker related to green-foot character of 817 meat hybrid chickens and application thereof |
| CN112746113A (en) * | 2021-01-29 | 2021-05-04 | 南昌师范学院 | Molecular marker related to chicken green foot character and application thereof |
| CN114214432A (en) * | 2021-12-31 | 2022-03-22 | 江苏北农大农牧科技有限公司 | Molecular marker for identifying melanin diffusion gene locus of chicken and application of molecular marker in assisted breeding |
Non-Patent Citations (2)
| Title |
|---|
| A high-density SNP-based linkage map of the chicken genome reveals sequence features correlated with recombination rate;Martien A.M.Groenen;《Genome Research》;第510-519页 * |
| 鸡毛脚性状的研究进展;聂庆华;《养禽与禽病防治》;第34-37页 * |
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