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Rapid detection method and application of single nucleotide polymorphism site of cattle cidec gene

A single nucleotide polymorphism, cattle technology, applied in the field of molecular genetics, can solve the problems of underdeveloped hindquarters, the speed of fattening and weight gain cannot keep up, and the meat production is less.

Inactive Publication Date: 2016-08-17
NORTHWEST A & F UNIV
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Problems solved by technology

The main disadvantages are that the hindquarters are underdeveloped, less meat is produced, the speed of fattening and weight gain cannot keep up with foreign specialized beef cattle breeds, and the output of high-quality and high-grade meat is less

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  • Rapid detection method and application of single nucleotide polymorphism site of cattle cidec gene
  • Rapid detection method and application of single nucleotide polymorphism site of cattle cidec gene
  • Rapid detection method and application of single nucleotide polymorphism site of cattle cidec gene

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Embodiment Construction

[0034] The present invention utilizes the PCR-RFLP method to detect the single nucleotide polymorphisms at the -974th, -956th, and -501st positions of the cattle CIDEC gene. The invention is explained rather than limited.

[0035] a. Design of PCR primers for cattle CIDEC gene containing 5-terminal flanking region

[0036] Using the bovine (AC_000179.1) sequence published by NCBI as a reference, Primer5.0 was used to design PCR primers capable of amplifying the 5-terminal flanking region of the CIDEC gene in cattle.

[0037] The primers for amplifying the 5-terminal flanking region are:

[0038] Upstream primer: aggagaacatttcagggtg 19bp;

[0039] Downstream primer: gcagtggaagttgggag 17bp.

[0040] The cattle genome was amplified with the above primers to amplify the fragment containing the 5-terminal flanking region of the cattle CIDEC gene. After sequencing and identification of the amplified fragment, a total of ten mutation sites were found: AC_000179.1:g.-501G>A; -546T...

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Abstract

The invention discloses a method for detecting polymorphism of multiple single nucleotides in a 5' control region of a cattle CIDEC gene. The method comprises the following steps of: with a to-be-detected cattle complete genome DAN containing a CIDEC gene as a template and primer pairs P (P1, P2 and P3) as primers, performing amplification on the cattle CIDEC gene; respectively digesting PCR amplification products with restriction enzymes such as HaeIII, AccI and HaeIII, then carrying out agarose gel electrophoresis on a digested amplified fragment, and identifying polymorphism of single nucleotides at a 974th site, a 956th site and a 501st site of the CIDEC gene of the cattle according to results of agarose gel electrophoresis, wherein polymorphism of the single nucleotide at the 956th site is also the polymorphism of single nucleotides at a 841st site, a 763rd site, 727th site and a 546th site, the polymorphism of the single nucleotide at the 501st site is also the polymorphism of a single nucleotide at a 643rd site owning to complete linkage, and thus polymorphism information of nine sites can be obtained by detecting polymorphism of three sites. The detection method provided by the invention lays a foundation for establishing a relation between the polymorphism of single nucleotides of the CIDEC gene and growth traits, so as to be convenient for marker assisted selection (MAS) on the growth traits for cattle beef in China, and a cattle population with excellent genetic resources can be quickly established.

Description

technical field [0001] The invention belongs to the field of molecular genetics and relates to the detection of gene single nucleotide polymorphism (SNP), in particular to a method and application for detecting the single nucleotide polymorphism of cattle CIDEC gene. Background technique [0002] DNA molecular markers can be used for the construction of livestock gene maps, marker-assisted protection of genetic resources, prediction of livestock heterosis, identification of individuals or breeds, genetic differentiation and analysis of kinship. With the in-depth development of the livestock genome project, livestock molecular marker breeding has become the main tool of livestock genetics and breeding and has brought huge economic value. [0003] Marker-assisted selection has gone through three stages of development: the first stage is the genetic analysis among various traits of livestock; the second stage is the stage of protein (enzyme) markers for quantitative traits; the...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 陈宏王璟曹修凯潘虹滑留帅蓝贤勇胡沈荣雷初超
Owner NORTHWEST A & F UNIV
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