Detection method for single nucleotide polymorphism of cattle PPARbeta gene and molecular breeding method
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A technology of single nucleotide polymorphism and detection method, which is applied in the field of detection of single nucleotide polymorphism of cattle PPARβ gene
Active Publication Date: 2016-05-04
XINYANG NORMAL UNIVERSITY
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There is no literature report on the genetic variation of PPARβ gene in Chinese native yellow cattle
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Embodiment 1
[0045] Example 1: Detection method of single nucleotide polymorphism of cattle PPARβ gene
[0046] 1. Design of PCR primers for cattle PPARβ gene
[0047] Taking the bovine PPARβ (AC_000180.1) sequence published by NCBI as a reference, the 195bp gene fragment of PPARβ gene from 71594bp to 71788bp Figure 4 and SEQ ID NO:5. Use Primer5.0 to design PCR primers that can amplify the 71616th region of the cattle PPARβ gene, and the primer sequences are as follows:
[0048] P3 (upstream primer): 5'-TCCTGTCTTCCCTTTCGTCC-3'20bp, see SEQ ID NO: 3;
[0049] P4 (downstream primer): 5'-GGAGACAACTCGCCCAAGAT-3'20bp, see SEQ ID NO: 4;
[0050] Using P3 and P4 primers to carry out PCR amplification on the cattle genome, a 427bp gene fragment including the 71616th position of the cattle PPARβ gene (AC_000180.1 sequence) can be amplified. After sequencing and identification of the amplified fragments, after analysis, It was found that there was a SNP polymorphism at position 71616 of the PP...
Embodiment 2
[0111] Example 2: Molecular breeding method based on single nucleotide polymorphism of cattle PPARβ gene
[0112] Genotype data: genotypes recognized by PvuII (AG and GG)
[0113] Production data: Birth weight of Nanyang and Jiaxian red cattle, as well as body weight, body height, body oblique length, bust, ischial end width, and daily gain at 6, 12, 18, and 24-month-old cattle.
[0114] Correlation analysis model: Descriptive analysis of the data is performed first to determine whether there are outliers, and then the least squares analysis is used to correct the data; according to the characteristics of the data, the SPSS (17.0) software is used to analyze the effects of production traits among genotypes. A fixed model was used in the analysis of genotype effects:
[0115] Y ijkl =μ+BF i +Month j +G k +e ijkl
[0116] Among them: Y ijkl is the observed value of traits, μ is the overall mean, BF i is the fixed effect of the i-th variety and farm, Month j is the fixe...
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Abstract
The invention discloses a detection method for single nucleotide polymorphism of a cattle PPARbeta gene. The detection method comprises the steps that PCR amplification is performed on the cattle PPARbeta gene by taking to-be-detected cattle whole genomeDNA containing the PPARbeta gene as a template and taking a primer pair (P) as primers; a PCR amplification product is digested with a restriction enzyme (Pvu II), and unmodified polyacrylamidegel electrophoresis is performed on segments obtained through enzymedigestion; the polymorphism of the 71616 single nucleotide of the cattle PPARbeta gene is identified according to an electrophoresis result. The invention further relates to a method for constructing a cattle molecular breedingindex system by means of the single nucleotide polymorphism of the PPARbeta gene. Due to the fact that the PPARbeta gene has the important biological function, the gene single nucleotide polymorphism has the significant influence on the weight, the withers height and the ischium end width of a cattle in the early stage and can be used for cattle beef and molecular marker-assisted selection of growth characteristics, and then a cattle population which has excellent genetic resources can be quickly constructed.
Description
technical field [0001] The invention relates to a method for detecting gene single nucleotide polymorphism and a molecular breeding method, in particular to a method for detecting single nucleotide polymorphism of cattle PPARβ gene and a method for molecular breeding. Background technique [0002] SNP (Single Nucleotide Polymorphism, SNP) mainly refers to the polymorphism of DNA sequence caused by the variation of a single nucleotide at a specific site on the genome level. The polymorphism shown by SNP is caused by the conversion or transversion of a single base. In genomic DNA, any base may be mutated. Therefore, SNPs may exist in both gene sequences and non-coding sequences. SNPs occur most frequently on CG-rich sequences, and most of them are cytosine C converted to thymine T, because C in CG will spontaneously deaminate to thymine after methylation. [0003] As a third-generation molecular genetic marker, SNP has the following characteristics: (1) High density. SNP is...
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