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Chicken laying trait related molecular marker and application thereof in chicken breeding

A molecular marker and trait technology, applied in the field of molecular genetics, which can solve the problems of affecting transcription initiation and gene expression, etc.

Active Publication Date: 2017-12-15
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, transcription is regulated by the sequence of the 5' regulatory region, and its base mutation usually affects the initiation of transcription, thereby affecting gene expression

Method used

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  • Chicken laying trait related molecular marker and application thereof in chicken breeding
  • Chicken laying trait related molecular marker and application thereof in chicken breeding
  • Chicken laying trait related molecular marker and application thereof in chicken breeding

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 Chicken miR-1b-3p 5' regulatory region sequence alignment and polymorphic site analysis

[0022] 1. Test material

[0023] 48 Recessive White Locke Chickens (Shandong Jihua Poultry Breeding Co., Ltd.), 48 Jining Hundred Day Chickens (Jining Datang Hundred Day Chicken Breeding Farm), 48 Xinyang Brown Chickens (Shanghai Poultry Breeding Co., Ltd.), random Sampling, blood collection from wing veins, genome extraction, and storage at -20°C.

[0024] 2. Test method

[0025] 2.1 Primer design

[0026] The primer P-miR-1b-3p was designed according to the published red jungle fowl sequence (GenBank Accession NC_006110). Sign in for the sequence design of the red jungle fowl.

[0027] 2.2 PCR amplification

[0028] Randomly select 48 genomes of Recessive White Rock Chicken, Jining Hundred Day Chicken, and Xinyang Brown Chicken, and use primer P-miR-1b-3p for PCR amplification. See Table 1 for primers, 20 μL of reaction system, including 1 μL of genomic DNA (50-10...

Embodiment 2

[0034] Example 2 Association analysis of chicken miR-1b-3p5'regulatory region polymorphism and age at first lay and egg production

[0035] 1 test material

[0036] Randomly selected 60 Jining hundred-day chickens (Jining Datang Hundred-day Chicken Breeding Farm), 49 Xinyang brown chickens (Shanghai Poultry Breeding Co., Ltd.) and 438 recessive White Rock chickens with egg production records (Shandong Jihua Poultry Breeders Ltd). All of the above were randomly sampled, blood was collected from the wing vein, and the genome was extracted and stored at -20°C.

[0037] 2 test method

[0038] 2.1 Genotyping of KASP SNP loci

[0039] KASP (Kompetitive Allele Specific PCR), that is, competitive allele-specific PCR.

[0040] Three primers were designed for the SNP / Indel site (the primer information is shown in Table 2), two forward specific primers and one reverse universal primer, and the two forward primers corresponded to the two fluorescent signals of FAM and HEX respectively...

Embodiment 3

[0066] Example 3 Effect of chicken miR-1b-3p 5'regulatory region polymorphic site on gene expression

[0067] 1 test material

[0068] Healthy Hai-Line brown chickens (3-5) in the peak egg-laying period (42w) of the farm in Linxi Village, Tai'an City were randomly sampled, and follicle tissues were collected for cell isolation and culture.

[0069] 2 test method

[0070] 2.1 Construction of wild-type and mutant luciferase expression vectors at the polymorphic site of miR-1b-3p 5′ regulatory region

[0071] 1) Select miR-1b-3p 5′ regulatory region -734 wild-type individuals in the recessive Bailock population, use their DNA as a template to obtain wt-miR-1b-3p, and design primers for fluorescent reporter gene detection, by Upstream - 857bp was amplified to downstream 141bp, the fragment length was 1013bp (including restriction site and protection base), its nucleotide sequence was shown in SEQ ID NO: 6, and the primer sequences used were shown in Table 5.

[0072] Table 5 Pr...

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Abstract

The invention belongs to the field of molecular genetics and particularly relates to a molecular marking method of a mutant site related to a 5' control region of a chicken miR-1b-3p gene and chicken sexual maturity and laying number traits and an application thereof in chicken breeding. The mutant site is a -734(G is greater than A) site. Genetic typing among three different groups is performed and the laying trait associated analysis is performed by means of a KASP technology, and the result shows that the site is extremely related to the laying start age and egg numbers in 32 weeks and 48 weeks (p is smaller than 0.01), and the laying start ages of genes AA and AG are extremely obviously earlier than that of GG gene, the egg numbers of the genes AA and AG in 32 weeks is extremely obviously greater than that of the gene GG but the egg number of the gene AA in 48 weeks is extremely obviously greater than those of the genes AG and GG. By detecting the molecular marker related to the laying trait, egg varieties / strains which are early to mature and high in yield are bred favorably, and favorable help is brought to local chicken breeding work.

Description

technical field [0001] The invention relates to the field of molecular genetics, and specifically provides a molecular marker related to chicken egg-laying traits and its application in chicken breeding. Background technique [0002] MicroRNAs (miRNAs) are a class of single-stranded non-coding small RNAs of about 18-25 nucleotides that widely exist in organisms. miRNAs regulate mRNA expression levels after transcription through specific complementary pairing with the 3'UTR of target genes. Inhibits translation of post-transcriptional proteins (Beilharz et al, 2010). MiR-1b-3p is expressed in skeletal muscle, cardiomyocytes, and chondrocytes of Xenopus laevis, zebrafish, mouse, and chicken (Townley et al., 2010, Vey et al., 2010, Dylan et al ., 2008, Kumi et al., 2010). Involved in cell proliferation and differentiation, and related to human ovarian cancer, breast cancer, diabetes and other diseases (Peng et al., 2015, Lin et al., 2015, Liu et al., 2015, Anquan et al., 2017...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156C12Q2600/158
Inventor 康丽李建波柯茂林段景德姜运良唐辉
Owner SHANDONG AGRICULTURAL UNIVERSITY
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