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Molecular marker related to sedimentary character of pork fat, and application thereof

A technology of molecular markers and pig fat, which is applied in the determination/inspection of microorganisms, DNA preparation, DNA/RNA fragments, etc., can solve the problems of lack of functional main effect genes and molecular markers, shorten the generation interval and speed up the breeding process Effect

Inactive Publication Date: 2013-12-18
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, in the practice of molecular breeding of pigs, there is still a lack of major genes and molecular markers with clear functions and significant effects.

Method used

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  • Molecular marker related to sedimentary character of pork fat, and application thereof
  • Molecular marker related to sedimentary character of pork fat, and application thereof
  • Molecular marker related to sedimentary character of pork fat, and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Cloning and sequencing of the porcine ECI1 gene of embodiment 1

[0042] Using the electronic cloning method, in the NCBI (http: / / www.ncbi.nlm.nih.gov / ) website, the ECI1 gene (NM_001919 and NM_010023) of humans and mice was homologously compared, and the porcine expressed sequence tag (EST) was searched. ), and then splice. After multiple alignments and splicing extensions, a sequence of 1401 bp was obtained, as shown in SEQ ID NO.2.

[0043] In order to verify the sequence obtained by electronic cloning, we designed two pairs of primers: primer pair ECI1-1 and primer pair ECI1-2 (see Table 1); among them, the primer pair ECI1-1 amplifies the 18th to 1165 bp of the sequence, and the expected product length 1148bp; ECI1-2 primer pair amplifies the 1037th to 1397th bp of the sequence, and the expected product length is 361 bp. Primers were diluted to 10ppm / μL with sterile double distilled water.

[0044] Liver tissue samples of Tibetan pigs were collected at the teach...

Embodiment 2

[0049] Example 2 Porcine ECI1 gene SNPs site screening

[0050] The porcine ECI1 gene mRNA sequence was BLATed in the porcine genome database of the UCSC website (http: / / genome.ucsc.edu / ) to obtain the genomic DNA sequence, and then three pairs of primers ECI1-3, ECI1-4 and ECI1-5 were designed (see Table 1), all exons and part of intron sequences of porcine ECI1 gene were amplified.

[0051] The Chinese local breeds "Tibetan pig" (collected from the teaching and practice ranch of Tibet Agriculture and Animal Husbandry College), "Southern Diannan small-eared pig" (collected from the resource conservation field of Diannan small-eared pig in Xishuangbanna Prefecture, Yunnan Province) and the introduced breed "Darkie pig" ( Beijing Shunyi pig farm) ear tissue samples, tissue genomic DNA was extracted by the phenol / chloroform method.

[0052] Using the genomic DNA of Tibetan pig, Diannan small-eared pig, and Yorkshire pig as templates, the porcine ECI1 gene sequence was amplified...

Embodiment 3

[0053] Example 3 Establishment of the PCR-RFLP detection method for the Intron3-C54T site of the porcine ECI1 gene

[0054] In order to quickly and conveniently detect the genotype of the Intron3-C54T mutation site of the porcine ECI1 gene, the DNAMAN software was used to analyze the restriction enzyme analysis and found that the mutation site can be recognized by the Bsm Ⅰ endonuclease (the recognition sequence is AATG); when the site When the point base is T, there is a Bsm Ⅰ restriction site; when the site is mutated to C, the Bsm Ⅰ restriction site disappears. For the Intron3-C54T mutation site of the porcine ECI1 gene, primers ECI1-6 (see Table 1) were designed to amplify the DNA sequence containing the Intron3-C54T site. The nucleotide sequence is shown in the sequence table SEQ ID NO.1. Wherein the 157th bp described in the sequence table SEQ ID NO.1 is a C / T mutation site.

[0055] Use ECI1-6 primers to perform PCR amplification on pig genomic DNA. The PCR reaction sy...

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Abstract

The invention provides a molecular marker, and particularly relates to cloning of a pig ECI1 gene segment and application as the molecular marker. A nucleotide sequence of the molecular marker is shown in SEQ ID NO.1; a C157-T157 base mutation is formed at the 157th bit, resulting in Bsm I-RFLP (restricted fragment length polymorphisms).

Description

technical field [0001] The invention relates to the field of animal molecular markers, in particular to the application of an ECI1 gene related to pig fat deposition traits as a molecular marker. Background technique [0002] Pig farming is the main body of my country's animal husbandry production, and its output value has always been the first in the total output value of animal husbandry for a long time. With the improvement of people's living standards, the demand for the quantity and quality of meat products continues to increase. How to breed pigs with high lean meat percentage and high intramuscular fat is a hot spot in current pig breeding work. [0003] my country is rich in pig breed resources, which provide good materials for pig breeding, and also provide good materials for the study of genes and molecular markers of important economic traits. Tibetan pig is a unique plateau pig breed in my country. It has a small body, thin skin, tender and delicious meat, rich ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12N15/10C12Q1/68
Inventor 张浩鲁云风强巴央宗李庆岗吴克亮张博王志秀
Owner CHINA AGRI UNIV
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