The method and application of detection of single nucleotide polymorphism of sheep pcnp gene by pcr-rflp

Abstract

The invention discloses a method for detecting single nucleotide polymorphism of a sheep PCNP (PEST-Containing Nuclear Protein) gene by using PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism) and application of the method. The method comprises the following steps: carrying out PCR amplification on a sheep PCNP gene segment by taking a sheep whole genome DNA to be measured as a template and taking a primer pair P as a primer; after a PCR amplification product is digested by a restriction enzyme SPhI, carrying out agarose gel electrophoresis on the digested amplified segment; authenticating the single nucleotide polymorphism of the 5019th site of the sheep PCNP gene according to an electrophoresis result. As a base mutation site is closely associated with the kidding number of sheep reproduction traits, the method is a method for detecting molecular genetic markers closely associated with the sheep reproduction traits at the DNA level, and can be used for auxiliary selection and molecular breeding of sheep and increasing the improved variety breeding speed of the sheep.

Description

technical field

[0001] The invention belongs to the field of molecular genetics, and relates to single nucleotide polymorphism (SNP) of sheep functional gene as molecular genetic marker, in particular to the single nucleotide polymorphism of sheep PCNP gene and a detection method thereof. Background technique

[0002] In sheep breeding, it is expected to achieve early selection and improve the accuracy of breeding values ​​through the selection of DNA markers that are closely related to reproductive traits and closely linked to quantitative traits, so as to obtain greater genetics in livestock and poultry breeding. progress.

[0003] Molecular breeding, that is, molecular marker-assisted selection breeding (Molecular Mark-Assist Selection, MAS), this technology is to use DNA molecular markers to select genetic resources or breeding materials, and to improve the comprehensive traits of livestock and poultry. The combination of biology and traditional genetics and breeding is...

Examples

Embodiment Construction

[0033] The present invention will be described in further detail below in conjunction with the accompanying drawings and embodiments, and the embodiments are intended to explain rather than limit the present invention.

[0034] The present invention uses PCNP gene conserved sequences to design primers to amplify the 686bp fragment of the second exon of PCNP gene, uses Hu sheep and small-tailed Han sheep genomes as templates, and conducts PCR amplification, and the amplified product is sequenced to find the single-point sequence of the amplified fragment. Nucleotide polymorphisms; carry out trait association analysis for discovered single nucleotide polymorphisms and provide detection methods, making the nucleotide polymorphism of PCNP gene a molecular genetic marker that can be quickly and easily detected, To provide a basis for accelerating the establishment of sheep populations with high-quality economic traits such as Hu sheep and Small-tailed Han sheep.

[0035] a. Detecti...

technical field

[0001] The invention belongs to the field of molecular genetics, and relates to single nucleotide polymorphism (SNP) of sheep functional gene as molecular genetic marker, in particular to the single nucleotide polymorphism of sheep PCNP gene and a detection method thereof. Background technique

[0002] In sheep breeding, it is expected to achieve early selection and improve the accuracy of breeding values ​​through the selection of DNA markers that are closely related to reproductive traits and closely linked to quantitative traits, so as to obtain greater genetics in livestock and poultry breeding. progress.

[0003] Molecular breeding, that is, molecular marker-assisted selection breeding (Molecular Mark-Assist Selection, MAS), this technology is to use DNA molecular markers to select genetic resources or breeding materials, and to improve the comprehensive traits of livestock and poultry. The combination of biology and traditional genetics and breeding is...

Examples

Embodiment Construction

[0033] The present invention will be described in further detail below in conjunction with the accompanying drawings and embodiments, and the embodiments are intended to explain rather than limit the present invention.

[0034] The present invention uses PCNP gene conserved sequences to design primers to amplify the 686bp fragment of the second exon of PCNP gene, uses Hu sheep and small-tailed Han sheep genomes as templates, and conducts PCR amplification, and the amplified product is sequenced to find the single-point sequence of the amplified fragment. Nucleotide polymorphisms; carry out trait association analysis for discovered single nucleotide polymorphisms and provide detection methods, making the nucleotide polymorphism of PCNP gene a molecular genetic marker that can be quickly and easily detected, To provide a basis for accelerating the establishment of sheep populations with high-quality economic traits such as Hu sheep and Small-tailed Han sheep.

[0035] a. Detecti...

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