A kind of ucp3 gene related to pig intramuscular fat traits, molecular marker and application thereof

A technology of molecular markers and intramuscular fat, which is applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve unclear problems and achieve the effect of improving accuracy and rapid genetic improvement

Active Publication Date: 2022-04-19
NANJING AGRICULTURAL UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Nevertheless, whether UCP3 is related to intramuscular fat and whether there are molecular markers in the gene that can be used for pig intramuscular fat selection is not clear

Method used

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  • A kind of ucp3 gene related to pig intramuscular fat traits, molecular marker and application thereof
  • A kind of ucp3 gene related to pig intramuscular fat traits, molecular marker and application thereof
  • A kind of ucp3 gene related to pig intramuscular fat traits, molecular marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Embodiment 1: Clone the full-length cDNA clone of porcine UCP3 gene

[0046] 1.1. Primer design

[0047] Take the NCBI database ( https: / / www.ncbi.nlm.nih.gov / ) The predicted porcine UCP3 gene mRNA sequence (NM_214049.1) seed sequence is used as a template to design amplification primers, wherein the 5'-RACE specific primers (GSPs) should satisfy the length of 23-28nt and the GC content of 50-70% between, and Universal Primer Mix ( The general primers designed by the RACE5' / 3'Kit kit have the conditions that the 3' end of Long Primer and Short Primer are not complementary, etc., and two primers that meet the above conditions are designed, namely GSP1 and NGSP1. GATTACGCCAAGCTT was added at the 'end to facilitate vector ligation and sequencing in subsequent experiments. The design of UCP3 gene 3'-RACE-specific primers was similar to that of 5'-RACE-specific primers. The designed primers were named GSP2 and NGSP2, respectively, and GATTACGCCAAGCTT was still added at...

Embodiment 2

[0055] Example 2: Analysis of pig UCP3 gene expression pattern

[0056] In order to determine the relationship between UCP3 expression pattern and fat deposition in pigs, the present invention randomly selected 3 Duroc×Meishan pig binary hybrid pigs, and collected heart, liver, lung, spleen, kidney, fat, and biceps femoris, At the same time, the skeletal muscle tissues of biceps femoris (fast muscle, white muscle) and soleus muscle (slow muscle, red muscle) of different muscle fiber types were collected from 5 experimental pigs. Utilize TRIzol reagent to carry out the extraction of total RNA (Invitrogen company product), utilize Prime Script TMReagents were used to synthesize the first strand of cDNA (products of TaKaRa Company). Then Real-time PCR was used to detect the mRNA expression level of UCP3 gene. Based on the cloned porcine UCP3 gene full-length cDNA nucleotide sequence SEQ ID NO: 1 as a reference, real-time PCR quantitative primers were designed. The quantitative ...

Embodiment 3

[0058] Example 3: Detection of genetic variation sites of pig UCP3 gene and identification of intramuscular fat-related molecular markers

[0059] 2.1. Experimental pigs and trait determination

[0060] The 279 (skin×du)×(length×big) quaternary commercial pigs involved in the present invention, the measured traits include carcass weight, backfat thickness, pH 45min , pH 24h , L* 45min 、a* 45min 、b* 45min , L* 24h 、a* 24h 、b* 24h , drip loss 24h , drip loss 48h , glycogen content, glucose content, glucose-6 phosphate content, lactic acid content, glycolytic potential, cooking loss, shear force. Finally, the rib backfat thickness was measured with an electronic vernier caliper, the pH value was measured with a portable pH meter (model: HI9125portable pH, Australia), the cooking loss was calculated using the loss rate after boiling for 10 minutes, and the shear force was measured with a domestic instrument. Meat color L*, a *, b* were measured with a portable colorimete...

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Abstract

The invention discloses a UCP3 gene, a molecular marker and its application related to pig intramuscular fat traits. The SNP molecular marker is a G / T mutation located at 2159bp of the sequence shown in SEQ ID NO: 6, and its genome position Located at site 8387309 from the 5' end of chromosome 9 of pig version 11.1. In the present invention, a mutation site that affects pig intramuscular fat content was identified in the gene by using the FastTarget target region sequencing technology in 279 test pigs, and a PCR-RsaI-RFLP gene rapid analysis for this site was established. type method; on this basis, the present invention has verified the application effect of improving intramuscular fat at this site in 500 test pigs, and the results show that the selection of favorable genotypes by this site can significantly improve the intramuscular fat of pigs. Fat content, the identified variant sites can be used for early, in vivo and rapid marker-assisted breeding of pig intramuscular fat.

Description

technical field [0001] The invention belongs to the field of livestock molecular breeding, and mainly relates to the screening and application of a molecular marker that can be used for marker-assisted selection of pig intramuscular fat traits. The molecular marker is located in the intron region of the UCP3 gene, and its genomic location is located at site 8387309 from the 5' end of chromosome 9 of pig version 11.1. Background technique [0002] Pig farming is one of the pillar industries of my country's animal husbandry industry. Pork is the main source of animal protein in people's lives. With the improvement of people's consumption level, the requirements for pork quality also increase accordingly. Therefore, the genetic improvement of pork quality has become a hot spot in current pig industry research. Meat quality (referred to as meat quality) can be evaluated by many technical indicators, such as meat color, backfat thickness, pH value, water resistance, intramuscula...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11C12N15/12
CPCC12Q1/6888C07K14/47C12Q2600/156C12Q2600/124
Inventor 吴望军李荣阳张增凯刘红林黄小国
Owner NANJING AGRICULTURAL UNIVERSITY
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