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Method for detecting single nucleotide polymorphism of cattle SH2B1 gene

A single nucleotide polymorphism, gene technology, applied in the field of molecular genetics

Inactive Publication Date: 2011-06-15
NORTHWEST A & F UNIV
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Problems solved by technology

At home and abroad (except SH2B1 and human obesity) there is no research on the genetic variation of SH2B1 gene in animals

Method used

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  • Method for detecting single nucleotide polymorphism of cattle SH2B1 gene
  • Method for detecting single nucleotide polymorphism of cattle SH2B1 gene
  • Method for detecting single nucleotide polymorphism of cattle SH2B1 gene

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Embodiment Construction

[0027] a. Design of PCR-RFLP primers for the first exon region of cattle SH2B1 gene

[0028] Taking the bovine (NW_001494303) sequence published by NCBI as a reference, Primer 5.0 was used to design PCR primers capable of amplifying the SNP site in the first exon region of the cattle SH2B1 gene. The primer sequences are as follows:

[0029] Upstream primer: cagcaactcc aactcctctg gc22;

[0030] Downstream primer: ctccccgggc cactccg17;

[0031] Among them, the upstream primer introduces mismatched bases, so as to form with the mutated SNP site and be recognized by endonucleases. Using the above primers to amplify the cattle genome, it is possible to amplify a 224bp gene fragment comprising the first exon region 2590bp to 2813bp of the cattle SH2B1 gene (NW_001494303 sequence), and the electrophoretic detection of the amplified fragment is as follows: figure 1As shown, wherein, lanes 1 to 6 are detection fragments, and lane M is Marker; after sequencing and identifying the ampl...

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Abstract

The invention discloses a method for detecting single nucleotide polymorphism of a cattle SH2B1 gene, which comprises the following steps: by using the whole genome DNA (deoxyribonucleic acid) of cattle to be detected, which contains an SH2B1 gene, as the template and using a primer pair P as the primers, carrying out PCR (polymerase chain reaction) amplification on the cattle SH2B1 gene; digesting the PCR amplification product by using a restriction endonuclease BglI, and carrying out agarose gel electrophoresis on the amplified segment which is subjected to enzyme digestion; and identifying the single nucleotide polymorphism of the 2795th site of the cattle SH2B1 gene according to the result of the agarose gel electrophoresis. Since the functions of the SH2B1 gene are related to four important growth traits, including body weight, daily weight gain, body oblique length and chest circumference, the method disclosed by the invention lays foundation for the establishment of the relationship between the SNP (single mucleotide polymorphism) of the SREBPlc gene and the growth traits, thereby facilitating the marker-assisted selection of growth traits for Chinese cattle beef and quickly establishing cattle populations with excellent genetic resources.

Description

technical field [0001] 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 2795th single nucleotide polymorphism of cattle SH2B1 gene. Background technique [0002] Gene polymorphism refers to the difference in genome sequence between different species or different individuals within the same species. These differences are caused by nucleotide changes in DNA alleles in chromosomes, mainly including base substitutions, insertions, deletions and duplications. Changes in sequence copy number. [0003] Single nucleotide polymorphism (Single Nucleotide Polymorphism, SNP) is a type of genetic marker system proposed by Lander (1996), a scholar at the Human Genome Research Center of the Massachusetts Institute of Technology. Polymorphism caused by acid (A / T / C / G) substitution. Its variant forms include: transversion, conversion, insertion and deletion, etc., w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 陈宏杨明娟屈炼刘俊霞蓝贤勇雷初朝
Owner NORTHWEST A & F UNIV
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