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pmel17 gene and genetic marker method related to melanin pigmentation of pectoral muscle of Xuefeng black-bone chicken

A technology of silky chicken and melanin, which is applied in the directions of genetic engineering, plant genetic improvement, botanical equipment and methods, etc., can solve the problem that genetic markers have not yet been reported, and achieve the effect of great application value and economic benefit.

Active Publication Date: 2019-06-21
湖南云飞凤农业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The research on DNA genetic markers, genetic diversity and related functional genes of Xuefeng black-bone chicken is still in its infancy, and there is no report on the genetic markers related to melanin deposition in Xuefeng black-bone chicken

Method used

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  • pmel17 gene and genetic marker method related to melanin pigmentation of pectoral muscle of Xuefeng black-bone chicken
  • pmel17 gene and genetic marker method related to melanin pigmentation of pectoral muscle of Xuefeng black-bone chicken
  • pmel17 gene and genetic marker method related to melanin pigmentation of pectoral muscle of Xuefeng black-bone chicken

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1 Xuefeng silky related gene SNP screening

[0051] 200 healthy Xuefeng black-bone chickens of the same age were selected for feeding experiment. The melanin content of these Xuefeng black-bone chicken breast muscles was determined, and the genomic DNA was extracted from samples with high melanin content and low melanin content samples.

[0052]According to the chicken pmel17 gene sequence in GenBank, Primer5.0 was used to design primers covering the entire sequence of exon 2 to exon 6 of the gene. The primer sequence is: 5'-ATCGCATCCCCGTCCCTTATC-3' (forward), 5'-GGACCCAACCATCCCTTGC -3' (reverse), synthesized by Hunan Qingke Biotechnology Co., Ltd. The genomic DNA of Xuefeng black-bone chicken was amplified with the above primers respectively.

[0053] PCR: 20 μl reaction system: 12 μl of 2×Master mix, 0.8 μl each of upstream and downstream primers (10 μmol / L), 1 μl of DNA template, and make up to the required volume with ultrapure water.

[0054] PCR amplifi...

Embodiment 2

[0058] Example 2 Verification of SNPs related to Xuefeng black-bone chicken breast muscle melanin in production practice

[0059] 44 healthy Xuefeng black-bone chickens with the same age and the same feeding and management conditions were selected for the feeding experiment. All the Xuefeng black-bone chickens tested were slaughtered at 120 days old, and the melanin content of breast muscle was determined.

[0060] Genomic DNA extracted from 44 Xuefeng black-bone chicken breast tissue samples was used to amplify using the primers in Example 1, and the SNP was detected according to the method in Example 1.

[0061] 2. PCR

[0062] The PCR reaction system is shown in the table below:

[0063] Table 1 PCR reaction system

[0064]

[0065] The PCR cycling parameters are as follows:

[0066] 34 cycles: 95°C, 5min; 95°C, 30s; 62.6°C, 30s, 72°C, 30s; 72°C, 5min

[0067] 2. Genotype frequency analysis of pmel17 gene SNP site

[0068] The genotype frequency and allele frequenc...

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Abstract

The invention discloses a pmel17 gene and a genetic marker used as a melanin character of the pectoral muscle of Xuefeng black-bone chicken. By screening the pmel17 gene polymorphism of Xuefeng black-bone chicken and measuring the melanin content of the pectoral muscle of Xuefeng black-bone chicken, and then analyzing the association between the pmel17 gene polymorphism and the melanin character to find that the pmel17 gene of Xuefeng black-bone chicken has a G / C base mutation at 185bp as shown in a sequence table, and the melanin content of the pectoral muscle of GG genotype Xuefeng black-bone chicken is significantly higher than other genotypes. The invention provides a molecular marker for marker-assisted selection of the melanin character of the pectoral muscle of Xuefeng black-bone chicken.

Description

technical field [0001] The invention belongs to the field of molecular biotechnology and breeding of Xuefeng black-bone chicken, mainly to measure the genetic markers related to the deposition of breast muscle melanin, specifically, to measure the polymorphism in the pmel17 gene, that is, to measure the polymorphism in the pmel17 gene, which is related to the Xuefeng black-bone chicken breast muscle Presence of single nucleotide polymorphism (Singlenucleotidepolymorphism, abbreviated as SNP) in melanin deposition-related pre-melanosome protein 17 (pmel17) gene. [0002] technical background [0003] Xuefeng black-bone chicken is a local breed of national genetic resources native to Huaihua City, Hunan Province. Its meat is fresh and tender, rich in nutrition, and has health benefits. The medicinal value of black-bone chicken is related to melanin. There is a record in "Compendium of Materia Medica" that "the darker the color, the better the medicine". Consumers also prefer to...

Claims

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Application Information

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IPC IPC(8): C12N15/12C12N15/11C12Q1/6888
Inventor 曲湘勇郭松长贺长青彭灿阳尹艳群
Owner 湖南云飞凤农业有限公司