Detection method for cattle RET (Reticulocyte) gene single nucleotide polymorphism site and detection kit and application thereof

A single nucleotide polymorphism and detection kit technology, applied in the field of biological genetic engineering, can solve problems such as disease in offspring, loss of household or feedlot, inability to screen for HSCR pathogenic mutation sites, etc., and achieves important economic value. low cost effect

Active Publication Date: 2018-11-20
XINYANG NORMAL UNIVERSITY
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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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  • Detection method for cattle RET (Reticulocyte) gene single nucleotide polymorphism site and detection kit and application thereof
  • Detection method for cattle RET (Reticulocyte) gene single nucleotide polymorphism site and detection kit and application thereof
  • Detection method for cattle RET (Reticulocyte) gene single nucleotide polymorphism site and detection kit and application thereof

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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-month-old red cattle.

[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 ...

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Abstract

The invention discloses a detection method for cattle RET (Reticulocyte) gene single nucleotide polymorphism sites. The detection method comprises the following steps: by taking whole genome DNA (Deoxyribonucleic Acid) of cattle to be detected with an RET gene as a template, a primer group P as a primer, carrying out PCR (Polymerase Chain Reaction) amplification on the cattle RET gene; digesting aPCR amplification product with a restriction enzyme MaeI (XspI), and carrying out native polyacrylamide gel electrophoresis on segments after digestion; according to polyacrylamide gel electrophoresis results, together with sequencing, judging the genotype of a single nucleotide polymorphism site in a 34690 position of the cattle RET gene. The invention further discloses application of the detection kit for cattle RET gene single nucleotide polymorphism sites in molecular breeding of cattle. Because of significant biological functions of the RET gene, the single nucleotide polymorphism of thegene has remarkable influence upon dip lengths, heights and chest circumferences of cattle at early stages and is applicable to auxiliary selection on molecular markers for meat and growth propertiesof the cattle, and furthermore cattle groups with excellent genetic resources can be rapidly established.

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 Applications(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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