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Method for identifying bull breeds with genetic markers of mononucleotides of Y chromosomes of Bos taurus

A Y-chromosome, single-nucleotide technology, applied in the field of molecular marker-assisted identification in introduction and breeding, can solve problems such as inability to distinguish varieties finely

Active Publication Date: 2017-07-07
NORTHWEST A & F UNIV
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Problems solved by technology

[0004] In view of the current research results, only a limited number of SNP sites have been found on common cattle (Bos taurus), and ordinary cattle can only be divided into two paternal origins, Y1 and Y2, and the problems of different breeds cannot be finely distinguished. The invention provides a method for identifying bull breeds by using common bovine Y chromosome single nucleotide genetic markers

Method used

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  • Method for identifying bull breeds with genetic markers of mononucleotides of Y chromosomes of Bos taurus
  • Method for identifying bull breeds with genetic markers of mononucleotides of Y chromosomes of Bos taurus
  • Method for identifying bull breeds with genetic markers of mononucleotides of Y chromosomes of Bos taurus

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example 1

[0030] Example 1. The method of identifying bull breeds using the common bovine Y chromosome single nucleotide genetic marker (SNP): DNA sequencing method

[0031] Taking the identification of Holstein bulls as an example, using the bovine Y chromosome reference genome sequence, select one or more of SNP1, SNP2 and SNP3, and extend a certain length on both sides, and use this as a template to design PCR amplification primers. Using the genomic DNA of the bull to be tested as a template and the DNA of a cow as a recessive control, PCR amplification was performed, and the amplified products were sequenced. Then use DNASTAR software to compare the sequencing results with the reference genome sequence to confirm whether the selected SNP site is a reference base or a mutation base. If it is a mutation base, the cow is a Holstein bull, otherwise it is not a Holstein bull. The specific steps are as follows , see figure 1 :

[0032] 1) extract the genomic DNA of the cattle to be tes...

example 2

[0042] Example 2, the method of identifying bull breeds using the common bovine Y chromosome single nucleotide genetic marker (SNP): SNP chip method

[0043] see figure 2 ,Specific steps are as follows:

[0044] 1) Design a 50bp single-stranded probe primer according to each SNP site in Table 1.

[0045] 2) Fix these primers on the chip in a certain order.

[0046] 3) The genomic DNA of the bovine sample to be tested is subjected to whole genome amplification.

[0047] 4) The amplified product is digested and fragmented with a random endonuclease.

[0048] 5) Hybridizing the DNA fragments with the chip, so that the genomic DNA digestion product can specifically bind to the probe.

[0049] 6) Two kinds of fluorescently labeled dideoxytriphosphate nucleotides (ddATP, ddGTP) are added for single base extension, and only the probes complementary to gDNA can be extended.

[0050] 7) The chip is scanned after cleaning.

[0051] 8) Detect the fluorescence intensity and wavelengt...

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Abstract

The invention discloses a method for identifying bull breeds with genetic markers of mononucleotides of Y chromosomes of Bos Taurus. In the method, according to a genome resequencing result, DNAs of Y chromosomes of main beef cattle and dairy cattle breeds are scanned, so that SNP (single nucleotide polymorphism) sites of specific Y chromosomes of Holstein cattle, Red Angus cattle, Charolais cattle, Gelvieh cattle, Limousin cattle, Simmental cattle, Han cattle and Hareford cattle breeds are discovered, and different breeds of bulls are identified in the DNA level according to the SNP sites. Therefore, the method can be effectively used for introduction and molecular mark assisted breeding of all the breeds, and a foundation is laid for fast establishment of a Bos Taurus population with excellent genetic resources.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for identifying bull breeds by using a common bovine Y chromosome single nucleotide genetic marker (SNP), which can be used for molecular marker-assisted identification of introduction and breeding. Background technique [0002] Single nucleotide polymorphism (SNP, single nucleotide polymorphism) is a phenomenon in which a single nucleotide differs in the allelic sequence of the genomic DNA of different individuals of the same species. The single-stranded DNA of the same length will have different conformations due to the difference in sequence or single base difference. The Human Genome Project has identified many SNPs, which are the most widespread type of variation in the genome of organisms, which are caused by insertions, deletions, transitions, and transversions of single nucleotides. [0003] Mammalian Y chromosome male-specific region (Male specific region, Y chromoso...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 姜雨李冉王喜宏曹春娜
Owner NORTHWEST A & F UNIV
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