SNP (Single Nucleotide Polymorphism) of related gene ELOVL3 influencing duck fatty acid composition

A technology of fatty acid and gene, applied in the field of poultry breeding and molecular biology, can solve the problem of less poultry meat, achieve the effect of improving breeding efficiency, increasing the content of long-chain polyunsaturated fatty acids in duck meat and meat quality

Active Publication Date: 2022-06-07
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are many studies on the fatty acid composition of other livestock at home and abroad, but there are few reports on the fatty acid composition and synthesis mechanism of poultry meat

Method used

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  • SNP (Single Nucleotide Polymorphism) of related gene ELOVL3 influencing duck fatty acid composition
  • SNP (Single Nucleotide Polymorphism) of related gene ELOVL3 influencing duck fatty acid composition
  • SNP (Single Nucleotide Polymorphism) of related gene ELOVL3 influencing duck fatty acid composition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Amplification of the intact cDNA sequence of duck polyunsaturated fatty acid composition related gene EVFLL3

[0032] Obtained the cDNA sequence of Duck chromosome 7LOVL3 (ENSAPLG00020013672) according to the Ensembl (http: / / asia.ensembl.org / ) database, and designed the amplification primers in the 5' UTR and 3' UTR regions, respectively.

[0033] The primer sequences are: 5'-GGACAGGCCAGAGGTCACT-3' (forward, SEQ ID NO.1), 5'-GAACAGGCGGGGCATGGTCACT-3' (reverse, SEQ ID NO.2), using cDNA obtained from reverse transcription of wild duck liver RNA as a template to amplify the sequence containing the gene's intact protein coding regions (CDs).

[0034] PCR: 20 μl reaction system: 2x PCR SuperMix 10 μl, upstream and downstream primers (10 μmol / L) 0.5 μl each, DNA template 1 μl, ultrapure water supplement 8 μl.

[0035] PCR amplification procedure: 94 °C prenaturation for 5 min; 94 °C denaturation 30s, 60 °C annealing 30s, 72 °C extension for 1 min, a total of 35 cycles;...

Embodiment 2

[0042] Example 2 Duck polyunsaturated fatty acid composition related gene EVFLL3 core promoter region confirmed

[0043] Obtained the upstream 2500bp sequence of Duck chromosome 7LOVL3 (ENSAPLG00020013672) from the Ensembl (http: / / asia.ensembl.org / ) database, designed fragment primers of different lengths using the online website Primer3 (https: / / bioinfo.ut.ee / primer3-0.4.0 / ), and added homologous recombination arms and enzymatic resection sites. Amplify missing fragments of different lengths, truncated primer sequences are:

[0044] Upstream primers (5'-3'):

[0045] ELOVL3-2095-F1:

[0046] ggcctaactggccggtaGGTACCAGTGAAGAGCTCCACGCACT(SEQ ID NO.5)

[0047] ELOVL3-1856-F2:

[0048]ggcctaactggccggtaGGTACCTGGGAAACAAGTCATGTCCA(SEQ ID NO.6)

[0049] ELOVL3-1518-F3:

[0050] ggcctaactggccggtaGGTACCCAGGGCAATTCCTCTAGCAT(SEQ ID NO.7)

[0051] ELOVL3-1249-F4:

[0052] ggcctaactggccggtaGGTACCCCCTGGGACAGCTCACAGTA(SEQ ID NO.8)

[0053] ELOVL3-765-F5:ggcctaactggccggtaGGTACCCACATCCTTCCATGCCACAT...

Embodiment 3

[0069] Example 3 SNP mutation effects associated with the above duck fatty acid composition related gene EVCL3 core promoter activity were validated

[0070] After obtaining the candidate mutation in the core promoter region, using the original expression plasmid as a template, the amplification primer of the single base mutation was introduced on the online website (https: / / crm.vazyme.com / cetool / singlepoint.html) for reverse amplification, and the primer sequence was:

[0071] 5'-ACgCAATTCTTGTAAAGAGACATACTATG-3' (forward, SEQ ID NO.13), 5'-CTTACAAGAATTGcGTAATAATACTTTGCTGAATTCTCCTGC-3',DpnI digestion of the amplification product, recombinant ligation reaction after removal of methylated template plasmids, and final product conversion, Coating and cloning identification (Mut.) II Fast Mutagenesis Kit V2,Vazyme bio,CHINA)。 The clonal liquid was sent to Beijing Qingke Biotechnology Co., Ltd. for sequencing, and the sequencing results were as follows Figure 5 to confirm that the sing...

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Abstract

The invention provides SNP (Single Nucleotide Polymorphism) of a related gene ELOVL3 influencing duck fatty acid composition and application of the related gene ELOVL3 in the aspects of meat duck breeding, meat quality identification and the like, the SNP in the application is located at the upstream-619bp of the ELOVL3 gene, and the genotype is AA, GG or GA. According to the present invention, GWAS analysis and cell test are combined to identify the SNP affecting the expression activity of the ELOVL3 gene in the ELOVL3 gene upstream region, further detection determines that the AA type individual has the higher polyunsaturated fatty acid carbon chain extension ability than the GG type individual, and the dominant genotype beneficial to increase of the fatty acid content of duck meat is selected to reserve the breed; the content of the long-chain polyunsaturated fatty acid in the duck meat and the meat quality can be rapidly improved, the progress of breeding improvement of high-quality ducks can be accelerated, and great promotion and objective economic benefits are brought to industrial development of the ducks.

Description

Technical field [0001] This application belongs to the field of molecular biology and poultry breeding. Specifically, the present application provides SNP and its applications affecting duck fatty acid composition-related genes LIKEL3. Background [0002] Meat and poultry is an important part of China's livestock and poultry production, with the characteristics of easy feeding and high production performance, and is the second largest consumer meat in the world. With the demand for healthy and diversified food in modern society, poultry products because of its nutritional balance, safety and health care and unique flavor advantages, the market share gradually increased, in the improvement of living standards, lifestyle changes and other factors, poultry product consumption structure continues to upgrade, consumers on poultry product quality safety and nutritional value more favored. [0003] Single nucleotide polymorphisms (SNPs) are caused by individual nucleotide variants at th...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156C12Q2600/124Y02P60/87
Inventor 侯卓成李晓琴
Owner CHINA AGRI UNIV
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