Method for detecting single nucleotide polymorphism of cattle AdPLA gene

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

Inactive Publication Date: 2010-12-22
NORTHWEST A & F UNIV
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Problems solved by technology

[0007] At present, the research on AdPLA gene is mostly on mice and humans, and ...

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

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Embodiment Construction

[0025] The present invention uses the PCR-RFLP method to detect the single nucleotide polymorphism of the 23076th synonymous codon mutation of the cattle AdPLA gene. The present invention will be further described in detail below, which is an explanation of the present invention rather than a limitation.

[0026] A. Design of PCR primers for the fourth exon region of cattle AdPLA gene

[0027] Taking the bovine (NC_007330.4) sequence published by NCBI as a reference, Primer 5.0 software was used to design PCR primers capable of amplifying the fourth exon region of the cattle AdPLA gene. The primer sequences are as follows:

[0028] Upstream primer: 5'-AGATACTCGCCATTGCCTCC-3' 20bp;

[0029] Downstream primer: 5'-GATGGGGGCACACTGAGGACCCACCTGATC-3' 30bp.

[0030] Among them, the third position from the 3' end of the downstream primer introduces a mismatched base G->A, thereby artificially constructing a FbaI restriction site TGATCA, which can be used to determine the genotype by PC...

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Abstract

The invention discloses a method for detecting the single nucleotide polymorphism of a cattle AdPLA gene, which comprises the following steps of: amplifying the cattle AdPLA gene through PCR (Polymerase Chain Reaction), wherein the DNA of a cattle genome to be testing, which comprises the AdPLA gene, is used as a template and a primer pair P is used as primers; digesting the PCR products by using restriction enzyme FbaI, and then performing agarose gel electrophoresis; and identifying the genetype of the 23075th bit of the cattle AdPLA gene according to the electrophoresis result. Because the functions of the AdPLA gene relate to the growth and body measurement traits of birth weight, weight, daily gain, oblique body length, chest circumference and the like, correlation analysis on genetype data and the traits shows that the oblique body length of 18-month and 24-month TT genetype individuals is obviously greater than that of CC genetype individuals (P is smaller than 0.05); the chest circumference of the 24-month TT genetype individuals is obviously greater than that of the CC genetype individuals (P is smaller than 0.05). The results indicate that the CC genetype can be used as a candidate molecular genetic marker for increasing the oblique body length and the chest circumference of cattle. The detection method can be applied to the marker assisted selection (MAS) of meat growth traits of Chinese cattle, and a cattle population with excellent genetic resources is quickly established.

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 23076th position SNP of cattle AdPLA 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. [0...

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 陈宏朱金龙张亚蓝贤勇侯飞雷初朝
Owner NORTHWEST A & F UNIV
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