Molecular marker for identifying Taihe silk chickens and silky fowls and application
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A technology of molecular markers and black-bone chickens, applied in the field of molecular biology, to achieve accurate identification, broad market application prospects, and easy operation
Active Publication Date: 2020-12-18
JIANGSU INST OF POULTRY SCI
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So far, there is no research report on the identification of silky c
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Embodiment 1
[0038] 1 Establishment of molecular detection method for silky chicken and bamboo silky chicken
[0039] 1.1 Primer design
[0040] The applicant carried out full-length sequencing of the mitochondrial genomes of the silky chicken population and the silky chicken population, and found that the two populations had a variation at 14945 bp, and designed primers to amplify the candidate site.
[0041] Forward primer (SEQ ID NO.1): 5'-TAACCAAAATCTCAAC-3'
[0042] Reverse primer (SEQ ID NO.2): 5'-GATAGTAATACCTGCG-3'
[0043] 1.2 PCR amplification
[0044] The PCR reaction system is:
[0045] 2×PCR Mix (Nanjing Novizan Biological Co., Ltd.) 25 μL, 10 μmol / L forward primer 1 μL, 50-100 μg / ml template DNA 2 μL, ultrapure water 21 μL.
[0046] The PCR reaction program was: 95°C for 5 minutes, 35 cycles (95°C for 30s, 60°C for 30s, 72°C for 60s), and 72°C for 10 minutes.
[0047] 1.3 Electrophoresis detection
[0048] The PCR product was detected by 1.3% agarose gel electrophoresis...
Embodiment 2
[0051] In this example, the molecular markers described in the present invention are used to carry out population expansion verification on silky chickens and bamboo silky chickens. The silky silky chicken comes from a national-level local chicken breed gene bank (Jiangsu), and the silky bamboo chicken comes from a large breeding enterprise. There are 30 of each species, and the total number is 60. The molecular markers are all GG type in silky chicken, and all are AA type in bamboo silky chicken. It was further verified that the SNP locus can effectively and reliably identify silky chicken and bamboo silky chicken (as shown in Table 1).
[0052] Table 1 is the genotype distribution of silky chicken and bamboo silky chicken detected by the molecular marker of the present invention.
[0053]
[0054] At the same time, only one SNP site was found in the whole mitochondrial genome, which is unique to silky chickens, and this site is located in the non-hypervariable region of...
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Abstract
The invention discloses a molecular marker for identifying Taihe silk chickens and silky fowls and application. The molecular marker is an SNP locus located at the 14945bp position of a mitochondrialgenome, has A/G polymorphism, is a silky fowl when the basic group of the position is A, and is a Taihe silk chicken when the basic group of the position is G. Primers SEQ ID NO.1 and SEQ ID NO.2 areused for amplifying a gene segment containing the molecule, then restriction enzyme HpaI is used for enzyme digestion of an amplification product, if a 690bp amplification segment is obtained, it proves that the chicken is the silky fowl, and if a 540bp amplification segment and a 150bp amplification segment are obtained, it proves that the chicken is the Taihe silk chicken. The method is simple to operate and reliable in result. The method can be used for quickly and accurately identifying the Taihe silk chickens and silky fowls, is easy to operate, low in cost and convenient to popularize and use in the grass roots level, and has wide market application prospects.
Description
technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a molecular marker for identifying silky chicken and silky bamboo chicken and its application. Background technique [0002] Silkie silkie is one of the oldest chicken breeds in my country, with historical records dating back to the 13th century, and it is also one of the few breeds listed as international standards in my country. The appearance of silky silky chicken is obviously different from that of other chicken breeds, and it has the standard "Shiquan" such as "mulberry crown, cherry head, green ears, beard, silk feathers, five claws, hairy feet, black skin, black bones and black meat". "Features have important ornamental functions. At the same time, silky chicken also has medicinal functions, and is the main raw material of the traditional Chinese medicine "Black-bone Chicken Baifeng Pills" for treating gynecological diseases. [0003] Because the s...
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