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Clone and use of molecular marker related to pig growth and meat quality traits

A technology of molecular markers and meat quality traits, applied in the field of livestock genetic engineering, can solve the problems of limited molecular markers affecting growth and meat quality

Inactive Publication Date: 2009-01-21
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] It can be seen from the above that although some progress has been made in the identification and application of candidate genes related to pig muscle growth and meat quality traits, the molecular markers that can really be used in breeding practices that affect growth and meat quality are still very limited. Related candidate genes and molecular markers are very necessary

Method used

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  • Clone and use of molecular marker related to pig growth and meat quality traits
  • Clone and use of molecular marker related to pig growth and meat quality traits
  • Clone and use of molecular marker related to pig growth and meat quality traits

Examples

Experimental program
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Effect test

Embodiment 1

[0031] Example 1 Cloning of JHDM1A gene

[0032] (1) Primer design:

[0033] Using human JHDM1A gene cDNA (GenBank accession number: NM_012308) as an information probe, using the BLAST tool in NCBI to perform homologous sequence screening in the GenBank pig EST database, a series of ESTs with a homology of more than 80% (the fragment length is greater than 100bp), use ENTREZ (http: / / www.ncbi.nlm.nih.gov / Web / Search / index.html) to query the corresponding sequences of these ESTs in NCBI, and then use the SeqMan program in DNAstar to construct pig EST-contigs; at the same time, using the human NM_012308 sequence to search for the porcine Trace-WGS sequence in GenBank, and according to the homology, it may be the partial sequence information of the corresponding exon of the corresponding porcine gene. Three pairs of primers P1, P2 and P3 were designed to amplify the JHDM1A gene cDNA sequence according to the human-mouse homologous sequence, EST splicing sequence and JHDM1A gene Tr...

Embodiment 2

[0051] Example 2 Physical location of JHDM1A gene:

[0052] (1) The primer sequence used for physical positioning is

[0053] P4: PL 5′-CCGCTGCGACCTTTGATGAC-3′ in intron 20

[0054] PR 5′-AAGGTGACGTTGGCGATGC-3′ in exon 21

[0055] The length of the fragment amplified by the primers is 498bp.

[0056] (2) Experimental materials for physical positioning

[0057] The pig×rodent somatic cell hybrid panel (SCHP) was used for chromosome region mapping, and the pig radiation hybrid panel (INRA-Minnesota porcine radiation hybrid panel, IMpRH) co-constructed by the University of Minnesota, USA was used for chromosome accuracy. Positioning, two sets of somatic cell hybrid plates were kindly donated by Dr. Martin Yerle (Laboratoire de Génétique Cellulaire, INRA, Castanet-Tolosan, France).

[0058] The radiation dose used by IMpRH is 7,000-rad. IMpRH includes 118 pig × hamster radiation hybrid cell lines, as well as hamster and pig genomic DNA positive controls, the identificatio...

Embodiment 3

[0063] Embodiment 3PCR-RFLP diagnostic method is established

[0064] (1) Primer sequence

[0065] P5: PL 5′-CCCAGACTGAGCAGGAACC-3′

[0066] PR 5′-GGACACCACGAGGAGAACC-3′

[0067] The length of the fragment amplified by the primers is 453bp.

[0068] (2) PCR amplification conditions

[0069] The total volume of the PCR reaction is 20μl, including about 100ng of porcine genomic DNA, containing 1×buffer (Promega), 1.5mmol / L MgCl 2 , the final concentration of dNTP is 150 μmol / L, the final concentration of primers is 0.2 μmol / L, and 2U Taq DNA polymerase (purchased from Promega Company). The PCR amplification program was: 94°C for 5min, then cycled 34 times at 94°C for 45s, 57°C for 45s, 72°C for 30s, and finally extended at 72°C for 5min. PCR reaction products were detected by 2% agarose gel electrophoresis and photographed.

[0070] (3) RFLP detection conditions

[0071] The volume of PCR product digestion reaction is 10μl, including 1μl of 10×buffer, 3-5μl of PCR pro...

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Abstract

The invention belongs to the livestock genetic engineering technical field, in particular relating to a molecular marker related to porcine growth and meat quality traits and used as a porcine marker auxiliary selection as well as an application thereof. The molecular marker is obtained through JHDM1A gene clone, and a cDNA sequence of the molecular marker is shown in a sequence table SEQ ID NO.1. NO. 213bp of a sequence table SEQ ID NO.2 has a base substitution of C213-G213, and the base substitution causes MboI-RFLP restriction enzyme polymorphism. The invention also discloses a primer used in partial cDNA sequences of an amplified JHDM1A gene and a method for polymorphism detection, and provides the novel molecular marker for porcine marker auxiliary selection.

Description

technical field [0001] The invention belongs to the technical field of livestock genetic engineering, and in particular relates to the cloning and application of molecular markers related to pig growth and meat quality traits. Background technique [0002] Pork is the main source of animal protein. With the development of economy and the improvement of people's living standards, people's demand for pork is constantly increasing in quantity and quality. Breeding pigs with fast growth and good meat quality is an effective way to meet the growing demand of consumers for pork. Breeding and producing breeding pigs with fast growth and high muscle quality has become an important trend in the development of my country's pig industry. [0003] Both growth traits and meat quality traits belong to quantitative traits and are controlled by polygenes. In the improvement of growth traits, in the past nearly 30 years, through the selection of phenotypes such as 100Kg body weight age and...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/12C12N15/10C12N15/11
Inventor 刘榜彭勇波樊斌余梅朱猛进徐学文彭中镇
Owner HUAZHONG AGRI UNIV