Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

A method for detecting recessive lethal mutation of bovine apaf1 gene

A lethal mutation and gene technology, applied in the field of molecular biology, can solve the problems of high cost, achieve high accuracy, improve the efficiency of species conservation, and reduce costs

Active Publication Date: 2019-12-06
CHINA AGRI UNIV +1
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method is not only expensive due to the dependence on microarray detection, but also needs to establish a genotype database of the reference population in advance; in addition, because it is essentially an indirect inference of the genotype of the disease-causing mutation site using the surrounding SNP markers, the result is not accurate. 100% accurate

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A method for detecting recessive lethal mutation of bovine apaf1 gene
  • A method for detecting recessive lethal mutation of bovine apaf1 gene
  • A method for detecting recessive lethal mutation of bovine apaf1 gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Establishment of Pyrosequencing Method for Detecting Recessive Lethal Mutation of Bovine APAF1 Gene

[0033] 1. Genomic DNA extraction

[0034] Genomic DNA was extracted from frozen bovine semen by high-salt method.

[0035] 2. Primer screening and PCR amplification

[0036] The target DNA sequence (UMD3.1:5:63150200:63150600:1) was obtained from the ensembl database (http: / / asia.ensembl.org / ), and PCR amplification primers and sequencing primers were designed. The position of the following primers and sequencing sequences on the bovine APAF1 gene sequence is shown in figure 1 .

[0037] Table 1 Primer combination for detection of recessive lethal mutation of bovine APAF1 gene

[0038]

[0039]

[0040] Among them, the upstream primer (APAF1-F) and the downstream primer (APAF1-R) will amplify the APAF1 gene fragment containing the recessive lethal mutation site; the sequence of the universal primer (APAF1-U) and the sequence of the downstream primer ...

Embodiment 2

[0058] The clinical application of embodiment 2 inventive method

[0059] Using the method described in Example 1 of the present invention, 152 Holstein bulls in the Beijing Dairy Center were clinically tested, and a total of 11 lethal mutation carriers were found, accounting for 7.2% of the carriers. Based on the test results, a selection and mating plan for these bulls was formulated, thereby gradually eliminating carriers of harmful genes and significantly improving the efficiency of selection.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides a method for detecting bovine APAF1 gene recessive lethal mutation, and relates to the field of molecular biology. The method is based on pyrosequencing techniques and is implemented by the aid of a pair of amplification primers, a universal primer and a sequencing primer. Nucleotide sequences are shown as SEQ ID No.1, 2, 3, and 4, single nucleotide polymorphism of 63150400bp sites of bovine chromosomes 5 is analyzed, as discovered in results, peaks are available at C base positions of wild type homozygotes of bovine APAF1 genes and are unavailable at T base positions of the wild type homozygotes of the bovine APAF1 genes, but peaks are available at C base positions and T base positions of lethal mutation carriers. The method has the advantages that the method is easy to implement, high in sensitivity, accuracy and flux and low in detection cost, and has important application value in bovine breeding procedures, and the individual APAF1 gene recessive lethal mutation carriers can be quickly detected by the aid of the method.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a SNP molecular marker for detecting the bovine APAF1 genotype and a method for detecting the recessive lethal mutation of the bovine APAF1 gene. Background technique [0002] Single nucleotide polymorphism markers (SNPs) mainly refer to DNA sequence polymorphisms caused by single nucleotide mutations on the genome. Detection of genotype by detection of single nucleotide polymorphism markers is a method that has emerged in recent years. Molecular markers have been used in animal breeding for some time. Compared with traditional breeding methods, molecular breeding greatly speeds up the efficiency of breeding, saves breeding time, and enables breeders to continuously explore and select better animals at the molecular level. Livestock breeds. [0003] A variety of methods have been used for SNP detection. Currently, gene chip methods, DNA sequencing methods, mass spectr...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156
Inventor 张毅劳兰兰肖炜俞英唐韶青刘林王雅春孙东晓张胜利
Owner CHINA AGRI UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com