Method for detecting single nucleotide polymorphism site of cattle ret gene, detection kit and application thereof

A single nucleotide polymorphism, detection kit technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problem of offspring sickness, inability to screen for HSCR pathogenic mutation sites, household heads or feedlots Losses and other issues, to achieve important effects of economic value

Active Publication Date: 2021-07-09
XINYANG NORMAL UNIVERSITY
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  • Summary
  • Abstract
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  • Claims
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Problems solved by technology

However, in the selection of breeding stock, the recessive pathogenic mutation cannot be directly reflected in the appearance, so people cannot screen the HSCR pathogenic mutation site
Once an individual carrying a recessive disease-causing mutation is selected as a breeding stock, its offspring will be at risk of disease, which will bring losses to the head of the household or the farm to a certain extent

Method used

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  • Method for detecting single nucleotide polymorphism site of cattle ret gene, detection kit and application thereof
  • Method for detecting single nucleotide polymorphism site of cattle ret gene, detection kit and application thereof
  • Method for detecting single nucleotide polymorphism site of cattle ret gene, detection kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1 Amplification of Cattle RET Gene Sequence and Detection of Polymorphic Sites

[0048] 1.1 Design of PCR primers for cattle RET gene

[0049] Taking the bovine RET (AC_000185.1) sequence published by NCBI as a reference, the 219bp gene fragment of the RET gene from 34669bp to 34887bp is as follows figure 1 shown. Use Primer 5.0 to design a PCR primer pair that can amplify the 34690th region of the cattle RET gene. The primer sequences are as follows:

[0050] Upstream primer P3: 5'-TCTCTGGGCTGAGTGGGAAA-3';

[0051] Downstream primer P4: 5'-CAAGGAAGCCCACCAGTCAG-3';

[0052] Using P3 and P4 primers to carry out PCR amplification on the cattle genome, a 486bp gene fragment including the 34690th position of the cattle RET gene (AC_000185.1 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 34690 of the RET gene.

[0053] Since there is no natural...

Embodiment 2

[0112] Example 2 The application of the single nucleotide polymorphism site detection method of yellow cattle RET gene in molecular breeding

[0113] Genotype data: genotypes (AG, GG and AA) recognized by MaeI (XspI).

[0114] 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 months of age.

[0115] Correlation analysis model: First, describe and analyze the data to determine whether there are outliers, and then use the least square method to analyze and correct the data; according to the data characteristics, use SPSS (18.0) software to analyze the effects of production traits among genotypes. A fixed model was used in the analysis of genotype effects:

[0116] Y ijkl =μ+BF i +Month j +G k +e ijkl

[0117] 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, Mo...

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Abstract

The invention discloses cattle RET The detection method of gene single nucleotide polymorphism site comprises the following steps: to contain RET The whole genome DNA of the cattle to be tested is used as a template, and the primer pair P is used as a primer to amplify the cattle by PCR. RET Gene; with restriction endonuclease MaeI (XspI) Digest the PCR amplification product, and then perform non-denaturing polyacrylamide gel electrophoresis on the digested fragment; determine the yellow cattle according to the results of polyacrylamide gel electrophoresis combined with sequencing RET The genotype of the single nucleotide polymorphism site at position 34690 of the gene. The invention also discloses RET The application of the detection kit of single nucleotide polymorphism site in yellow cattle molecular breeding. because RET The gene has important biological functions. The single nucleotide polymorphism of the gene has a significant impact on the early body length, body height, and bust of cattle. It can be used for molecular marker-assisted selection of cattle and growth traits, and then quickly establish excellent genetic resources. cattle population.

Description

technical field [0001] The invention belongs to the technical field of biogenetic engineering, and in particular relates to a method for detecting a single nucleotide polymorphism site of a cattle RET gene, a detection kit and an application thereof. Background technique [0002] Cattle are one of the main livestock resources in my country, but now they are facing the pressure of insufficient provenance of excellent cattle breeds and poor genetic quality of the population. Traditional breeding methods are based on determining the phenotype of animals and using the genetic information of their parents, ancestors and other relatives in animal models to achieve genetic evaluation. Generally, it is assumed that traits are affected by genetic differences of many genes, so each gene is considered There are relatively small contributions to the trait, with the result that the estimates of the trait are inevitably somewhat biased. Molecular genetic markers are one of the fastest-gr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6858
CPCC12Q1/6858C12Q2531/113C12Q2565/125
Inventor 黄洁萍马云赵明辉张明明郝瑞杰魏雪锋
Owner XINYANG NORMAL UNIVERSITY
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