Method for detecting cattle Six6 gene single nucleotide polymorphism

A single nucleotide polymorphism and gene technology, applied in the field of molecular genetics, can solve problems such as blanks and lack of research

Inactive Publication Date: 2010-01-20
NORTHWEST A & F UNIV
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Problems solved by technology

At present, there is a lack of research in the field of genetic variation of the Six6 gene in Chinese cattle. The functional research of this gene locus and its genetic variat

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  • Method for detecting cattle Six6 gene single nucleotide polymorphism
  • Method for detecting cattle Six6 gene single nucleotide polymorphism
  • Method for detecting cattle Six6 gene single nucleotide polymorphism

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[0026] The present invention utilizes the RFLP-PCR method to detect the single nucleotide polymorphism that may result in a change in the conformation of the encoded protein due to the nonsense codon mutation at the 2015th position of the Six6 gene of cattle. The present invention is further described in detail below. It is to explain rather than limit the present invention.

[0027] a. Design of PCR primers for the region containing the stop codon of the Six6 gene of cattle

[0028] Taking the Hereford cattle (NC_007308) sequence published by NCBI as a reference, using Primer 5.0 to design PCR primers that can amplify the stop codon region of the cattle Six6 gene, the primer sequences are as follows:

[0029]Upstream primer: gggctgactg ctgggctta 19;

[0030] Downstream primer: agaccaagca acccagcg 18;

[0031] Amplification of the cattle genome with the above primers can amplify the gene fragment of 383bp comprising the stop codon region of the cattle Six6 gene (NC_007308 se...

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Abstract

The invention discloses a method for detecting cattle Six6 gene single nucleotide polymorphism, comprising the following steps: taking the to-be-detected cattle entire genome DNA as a template, taking primer pair P as primers, and carrying out PCR amplification on the cattle Six6 gene; after using restriction endonuclease Hhal to digest the PCR amplified product, carrying out polyacrylamide gel electrophoresis on the amplified fragment after enzyme cutting; identifying the single nucleotide polymorphism of the 2015th of the cattle Six6 gene according to result of the polyacrylamide gel electrophoresis. As the Six6 gene has the functions that relate to growth traits, such as daily gain, body depth, weight, body slant length, ischium width and the like, the detection method provided by the invention lays out a foundation for the establishment of the relation between SNP and growth traits of the Six6 gene, so that the invention can be used for marker-assisted selection (MAS) of growth traits for meat of cattle of China and quickly establishing cattle population with excellent genetic resources.

Description

[0001] biotechnology field [0002] The invention belongs to the field of molecular genetics and relates to the detection of gene single nucleotide polymorphism (SNP), in particular to a method for detecting the 2015th SNP of cattle Six6 gene. Background technique [0003] 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. [0...

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Application Information

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IPC IPC(8): C12Q1/68G01N27/447
Inventor 陈宏淮永涛蓝贤勇王璟马亮屈玉娇任刚赖新生胡沈荣雷初朝
Owner NORTHWEST A & F UNIV
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