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Method for detecting single nucleotide polymorphism of cattle chemerin gene

A single nucleotide polymorphism and detection method technology, which is applied in the direction of microbial determination/inspection, biochemical equipment and methods, etc., can solve the problems of no yellow cattle, etc., and achieve low-cost and high-precision effects

Inactive Publication Date: 2012-07-04
NORTHWEST A & F UNIV
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Problems solved by technology

There is only one study on genetic variation of chemerin gene in humans (Mussig K, StaigerH, Machicao F, Thamer C, Machann J, Schick F, Claussen CD, Stefan N, FritscheA, Haring HU. RARRES2, encoding the novel adipokine chemerin, is a genetic determinant of disproportionate regional body fat distribution: A comparative magnetic resonance imaging study. Metabolism. 2009; 58: 519-524.), but so far there has been no report on the genetic variation or SNP study of the cattle chemerin gene

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  • Method for detecting single nucleotide polymorphism of cattle chemerin gene
  • Method for detecting single nucleotide polymorphism of cattle chemerin gene
  • Method for detecting single nucleotide polymorphism of cattle chemerin gene

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Embodiment Construction

[0036] 1. Design of PCR primers for the region containing the ninth exon of cattle chemerin gene

[0037] Taking the bovine (NC_007302.4) sequence published by NCBI as a reference, Primer 5.0 was used to design PCR primers capable of amplifying the ninth exon region of the ox chemerin gene. The primer sequences are as follows:

[0038] Forward primer: 5'-CGGAGCCCACCAGACAGG-3'18nt;

[0039] Reverse primer: 5'-CAGCGGGCAGCAGCACAC-3'18nt;

[0040] Using the above primers to amplify the cattle genome, it is possible to amplify a 441bp gene fragment including the first exon region of the cattle chemerin gene (NC_007302.4 sequence) and part of the intron 750-1190bp; sequence the amplified fragment After identification, it was found that when the G mutation at the 868th bp (that is, the 12th position of the CDS region of the chemerin gene) was changed to A, the amino acid codon encoding the 4th nt was mutated from CTG to CTA, thereby forming a synonymous mutation of leucine; and Thi...

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Abstract

The invention discloses a method for detecting single nucleotide polymorphism of a cattle chemerin gene. Gene polymorphism comprises A or G base polymorphism in the 868th site of the cattle chemerin gene and G or A base polymorphism in the 1021st site of the cattle chemerin gene. The detection method for the single nucleotide polymorphism of the cattle chemerin gene comprises the following steps of: by using a cattle entire genome DNA (deoxyribonucleic acid) to be detected, containing a chemerin gene, as a template, and a primer pair (comprising P and newP) as primers, carrying out PCR (Polymerase Chain Reaction) amplification on the cattle chemerin gene; after a PCR amplified product is digested by using restriction enzyme Pvu II, carrying out agarose gel electrophoresis on an amplified fragment subjected to enzyme digestion; and identifying the single nucleotide polymorphism of the 868th site and the 1021st site of the cattle chemerin gene according to an agarose gel electrophoresisresult. The method is used for screening and detecting a molecular genetic marker which is closely related to cattle production trait on the basis of a DNA level, carrying out marker assisted selection (MAS) on Chinese cattle beef growth trait and quickly establishing cattle specie groups with favorable genetic resources.

Description

technical field [0001] The invention belongs to the field of molecular genetics, relates to the detection of gene single nucleotide polymorphism (SNP), in particular to a method for detecting the 868th and 1021st SNPs of cattle chemerin gene. Background technique [0002] Single nucleotide polymorphism (SNP) refers to the polymorphism caused by the substitution of a single nucleotide (A / T / C / G) in the genomic DNA sequence. Therefore, commonly referred to as SNPs include base substitutions, insertions, deletions, and changes in the copy number of repeated sequences. A SNP indicates that there is a nucleotide change at a certain position in the genome, which is mainly caused by the conversion or transversion of a single base; SNPs with conversion mutations account for about 2 / 3, and other SNPs are in similar level. The cytosine of the CpG dinucleotide is the most mutated site in the genome, most of which are methylated and can be spontaneously deaminated to form thymine. [...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 陈宏张亚朱金龙徐瑶蓝贤勇雷初朝
Owner NORTHWEST A & F UNIV
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