Genetic marker using pig MLC(myosin light chain)2 5' flanking promoter region SNP (single nucleotide polymorphism) as pig carcass traits and application

A technology of genetic markers and promoters, applied in the direction of DNA/RNA fragments, recombinant DNA technology, microbial determination/inspection, etc.

Inactive Publication Date: 2012-11-14
HUAZHONG AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the research on the SNP in the promoter

Method used

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  • Genetic marker using pig MLC(myosin light chain)2 5' flanking promoter region SNP (single nucleotide polymorphism) as pig carcass traits and application
  • Genetic marker using pig MLC(myosin light chain)2 5' flanking promoter region SNP (single nucleotide polymorphism) as pig carcass traits and application
  • Genetic marker using pig MLC(myosin light chain)2 5' flanking promoter region SNP (single nucleotide polymorphism) as pig carcass traits and application

Examples

Experimental program
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Effect test

Embodiment 1

[0023] Example 1 Utilize phenol extraction method to extract large white pig and Meishan pig blood total DNA

[0024] Add 0.5M ethylenediaminetetraacetic acid (EDTA, purchased from Wuhan Jiangrun Fine Chemical Co., Ltd.) to the fresh blood of large white pigs collected on the spot (from the experimental pig farm of Huazhong Agricultural University, which is a conventionally reported variety, the same below). As an anticoagulant (according to the volume ratio of blood volume 1 / 10), and shake well to prevent coagulation.

[0025] Leukapheresis:

[0026] (1) 4°C, 6000rpm, centrifuge for 10min, remove serum.

[0027] (2) Add 2 times the volume of double-distilled water, shake gently to precipitate for 10 minutes, and break the red blood cells.

[0028] (3) 4°C, 6000rpm, centrifuge for 10min, remove the upper red blood cell plasma.

[0029] (4) The precipitate was washed with 0.9% NaCl and shaken gently for 10 min.

[0030](5) 4°C, 6000rpm, centrifuge for 10min, discard the sup...

Embodiment 2

[0039] Example 2: Acquisition of a specific DNA fragment for the 5' flanking promoter region of pig MLC2 and establishment of a SNP detection method

[0040] The foreign ancestry pig "Dabai pig" and the Chinese local pig breed "Meishan pig" (from the experimental pig farm of Huazhong Agricultural University, which is a routinely reported variety) were selected as the experimental materials, and the mRNA sequence of the pig MLC2 gene (accession number AY754870) was used as the seed , compared in the pig genome, and designed the following primers according to its 5' flanking sequence:

[0041] Forward primer F: 5'TATCCTTCTGTCCTCCTG 3'

[0042] Reverse primer R: 5'CTCTATGACCTTGGACTTG 3'

[0043] Using large white pig (foreign blood-related pig breed) and Meishan pig (Chinese local pig breed) blood genome DNA as templates, PCR amplification was carried out with the above primers (see figure 1 ).

[0044] The PCR reaction system is shown in Table 1.

[0045] Table 1 PCR reacti...

Embodiment 3

[0052] Example 3: Association analysis and application of genetic markers cloned in the present invention and pig carcass traits

[0053] Selected 249 Large White × Meishan pigs established in 2000, 2003 and 2004 by the Key Laboratory of Pig Genetics and Breeding of the Ministry of Agriculture, Huazhong Agricultural University, Wuhan, Hubei Province, China 2 (Liu et al.Association of MYF5 and MYOD1 gene polymorphisms and meat quality traits in Large White×Meishan F 2 pig populations.Biochem Genet.2008, 46:720-732) generation resource population as experimental materials According to the typing results of HinfI-RFLP method, the relationship between different genotypes of HinfI-RFLP in the 5' flanking promoter region of pig MLC2 gene and pig carcass traits was analyzed relationship. The SAS statistical software glm program was used to perform single-marker analysis of variance, and the model was as follows:

[0054] Model Y ijk =μ+G i +S j +Y k +βcov ijk +e ijk

[0055...

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Abstract

The invention belongs to the technical field of pig molecular marker preparation and particularly relates to a genetic marker using pig MLC(myosin light chain)2 5' flanking promoter segments as pig carcass traits as well as a preparation method and application of the genetic marker. The genetic marker has the nucleotide sequence shown as SEQ ID NO (sequencer identifier number):1 and SEQ ID NO:2 in a sequence table. One A/G mutation and one G/A mutation respectively exist at the 53bp part of the sequence shown as SEQ ID NO:1 and SEQ ID NO:2, and the polymorphism of DraIII-RFLP is caused by the allele mutation. The genetic marker is used for carrying out pig carcass traits correlation analysis on large white pig and meishan pig F2 generations, and the transcriptional activity of promoters in different genotypes is detected. The invention also discloses a parting detection method of the genetic marker, and the novel genetic marker and the detection method are provided for the pig carcass traits molecular marker auxiliary selection.

Description

technical field [0001] The invention relates to the fields of pig breeding and pig molecular marker-assisted selection, in particular to the cloning of a gene MLC2 5' flanking promoter fragment related to pig carcass traits and its application as a genetic marker for pig carcass traits. Background technique [0002] It has been thousands of years since human beings have been unconsciously or consciously selecting the production performance of livestock and poultry, while the discussion from phenotype selection practice to genotype selection theory has only been done in recent decades. When selecting the production performance of livestock and poultry, it mainly relies on the markers of its target traits, while the previous biochemical immune markers and cytogenetic markers have limited detection sites and poor polymorphisms. After the 1980s, the birth and gradual improvement of molecular genetic marker technology enabled animal breeders to quickly apply DNA molecular markers...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/113
Inventor 刘敏徐德全孙小瑞刘倩夏晓亮熊远著
Owner HUAZHONG AGRI UNIV
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