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Maternal genetic marker for identification of domestic cattle of bos and identification method and application thereof

A genetic marker and maternity technology, applied in the fields of application, botanical equipment and methods, biochemical equipment and methods, etc., can solve the problems of inconvenience, inaccurate appearance identification and cluster analysis, etc., and achieve the effect of simple identification method

Inactive Publication Date: 2012-05-09
YUNNAN AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0008] The technical problem to be solved by the present invention is to overcome the inaccurate and inconvenient technical defects of the existing appearance identification and cluster analysis used in the identification of cattle of the genus Bovis, and its purpose is to provide a simple, accurate and effective identification method. Maternal genetic markers of cattle breeds in the genus Bos (Bos taurus), zebu (Bos indicus), large-fronted cattle (Bos frontalis), yaks (Bos grunniens) and their hybrids

Method used

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  • Maternal genetic marker for identification of domestic cattle of bos and identification method and application thereof
  • Maternal genetic marker for identification of domestic cattle of bos and identification method and application thereof

Examples

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Effect test

Embodiment 1

[0028] Embodiment 1 A group of maternal genetic markers for differentiating cattle of the genus Bovis of the present invention and preparation method thereof

[0029] The preparation steps are as follows:

[0030] (1) Design primers according to the full sequence of common yellow cattle mtDNA (in GenBank accession number: V00654), the primers are composed of forward primers and reverse primers, the DNA sequence of the forward primers is as SEQ ID No. 5, the DNA sequence of the reverse primer is shown in SEQ ID No.6.

[0031] The above primers are conservative primers, which can simultaneously amplify the mtDNA sequences of 4 cattle species: yak, large-faced cattle, zebu and common yellow cattle. The above primers were synthesized by Sangon Bioengineering (Shanghai) Company.

[0032] (2) Through the appearance identification and breeding records of the sample source individuals, it is confirmed to select Bos (Bos) yaks (directly sourced from Diqing Prefecture, Yunnan Province...

Embodiment 2

[0038] Embodiment 2 Differentiate the method for each family cattle of the genus Bovis and hybrid individuals thereof

[0039] The cattle of the genus Bovis are common yellow cattle, zebu, large-faced cattle and yaks, and the steps are:

[0040] (1) Extract bovine genomic DNA from the bovine blood of the bovine species to be identified respectively, and its extraction method is to extract by phenol / chloroform conventional method, " Genetic Engineering ", Lou Shilin, 2002, Science Press.

[0041] (2) Use the primers designed in Example 1 to perform PCR amplification on the genomic DNA of the four kinds of cattle extracted in step (1). PCR reaction system 25μL, the concentration of each component in the system is: 50ng template DNA, 1×Taq buffer 2.5mmol / L, 1.5mmol / L MgCl 2 , 2.5mmol / L dNTPs, 0.5mmol / LPCR primers, 2U TaqDNA polymerase (MBI), the PCR operation program is as follows: pre-denaturation at 94°C for 4min; 94°C for 45s, 56°C for 40s, 72°C for 40s, 35 cycles; finally T...

Embodiment 3

[0044] Example 3 Consistency comparison between the bovine species-specific deletion genetic markers screened by the present invention and the clustering results

[0045]The sequences of yaks, large-faced cattle, zebu and common cattle containing the sequence fragments of the present invention were downloaded from GenBank, including 322 yaks, 76 large-faced cattle, 391 Zebu, and 1511 common cattle, a total of 2300 samples. Outgroups of 7 buffalo sequences downloaded for cluster analysis. These sequences were edited and compared by DNAstar software (DNASTAR, Madison, WI), and sequence polymorphism analysis was performed by MEGA 2.1 software (KumarS.et al.2001 MEGA.Molecular Evolutionary Genetics Analysis Software.Release 2.0.Arizona State University, Tempe A.Z., USA) completed, cluster analysis does not consider missing variants. The haplotype NJ tree and MP tree were completed by PAUP software (Swofford D.L.2000 PAUP.phylogenetic analysis using parsimony and other methods.Rel...

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Abstract

The invention discloses maternal genetic markers for identification of domestic cattle of bos and an identification method and application thereof, belonging to the technical field of genetic marker-assisted selection. The maternal genetic marker for ordinary cattle comprises specific deletion gap249, the maternal genetic marker for zebu cattle comprises specific deletion gap191, and the maternal genetic marker for gayal and yaks comprises specific deletion gap179-186. In the identification method, the genetic markers of specific deletion of the three types of cattle are used for identification, and the results are completely consistent with cluster results. Therefore, the identification method can completely substitute the complex cluster analysis for convenient and effective identification of zebu cattle, ordinary cattle or gayal and yaks, and can overcome the inaccuracy of cattle appearance identification. The primers provided by the invention for simultaneous expansion of the DNA of the four types of cattle are shown in SEQ ID No. 5 and SEQ ID No. 6.

Description

technical field [0001] The invention belongs to the technical field of bovine genetic marker-assisted selection, and in particular relates to a group of matrilineal genetic markers capable of distinguishing purebred cattle and hybrid cattle, primers thereof, and methods and applications for distinguishing cattle breeds. Background technique [0002] In terms of phenotype, domestic cattle can be divided into two main types: humped zebu and humpless taurine. The two types are two independent species (Bosindicus and Bos taurus) of the genus Bos (Bos) from two independent domestications of aurochs (Bos primigenius) (Loftus R.T. et al. Evidence for two independent domestications of cattle.Proc . Natl. Acad. Sci. 1994, 91: 2757-2761). The Bovine genus also includes two domestic cattle species, Yak (Bos grunniens) and Gayal (Bos frontalis). [0003] Yak (Bos grunniens) is a rare domestic cattle species distributed in the Qinghai-Tibet Plateau and its adjacent alpine regions. Exi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/12C12N15/11C12N15/10
Inventor 苟潇毛华明杨舒黎王永奇鲁绍雄邓卫东吕佳琪韩树鑫刘兴勇王秀娟
Owner YUNNAN AGRICULTURAL UNIVERSITY
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