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Method for detection of sheep fecundity by utilizing GnRHR gene

A technology for genetic testing and fecundity, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the correlation analysis of polymorphic genotype and litter size, influence the fecundity of Small-tailed Han sheep and Dorset sheep , Unable to determine the GnRHR gene and other problems

Inactive Publication Date: 2014-04-23
刘武军 +1
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  • Abstract
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  • Application Information

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Problems solved by technology

However, this study did not conduct an association analysis between polymorphic genotypes and litter size, and it is impossible to determine whether the GnRHR gene is the main gene that affects the fertility of Small-tailed Han sheep and Dorset sheep

Method used

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  • Method for detection of sheep fecundity by utilizing GnRHR gene
  • Method for detection of sheep fecundity by utilizing GnRHR gene
  • Method for detection of sheep fecundity by utilizing GnRHR gene

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Experimental program
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Embodiment

[0022] 1. Genomic DNA extraction:

[0023] Collect 5 mL of jugular vein blood samples from freshly slaughtered Cele black sheep and Turpan black sheep, anticoagulate with sodium citrate, and synthesize exon 2 of the amplified GnRHR gene by referring to the "Guidelines for Molecular Cloning Experiments"; use phenol-chloroform extraction method to extract Genomic DNA from blood samples should be stored at -20°C.

[0024] (1) Thaw the blood sample, transfer 1000L to a 2.0ml centrifuge tube, add an equal volume of PBS buffer, shake gently to mix; centrifuge at 12000r / min for 10min, discard the supernatant, repeat the above steps until the supernatant is transparent and the precipitate is light yellow .

[0025] (2) Add 500 l of DNA extraction solution into the centrifuge tube, shake to make the blood cell pellet detach from the wall of the centrifuge tube, and bathe in water at 37°C for 1 hour.

[0026] (3) Add proteinase K to 10l (20mg / mL) and mix well, overnight at 55°C until ...

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Abstract

The invention provides a method for detection of sheep fecundity by utilizing a GnRHR gene, and is used for finding out a molecular marker obviously affecting the sheep fecundity. The method comprises the steps: selecting the GnRHR gene as a candidate gene of the sheep fecundity, utilizing a PCR-SSCP technology, detecting genetic polymorphism of a GnRHR gene exon 2 in a Cele black sheep breed and a Turpan black sheep breed, and analyzing and studying influence of gene mutation on a sheep fecundity trait. Eventually, gene mutation G230C of the GnRHR gene exon 2 region is found to be capable of reducing the sheep fecundity, and at the same time, through selecting individual sheep with the genotype of GG and eliminating individual sheep with the genotypes of GC and CC, the sheep fecundity can be selectively bred and improved.

Description

technical field [0001] The invention relates to molecular marker technology, a method for detecting sheep fecundity by using GnRHR gene and its application in breeding to improve sheep fecundity, and has important production value. Background technique [0002] As one of the five major pastoral areas in my country, Xinjiang had a population of 41.6152 million sheep and goats in 2013. It has abundant sheep breed resources, especially the excellent local sheep breeds of Cele black sheep and Turpan black sheep. It is an effective means and fundamental method to meet the growing needs of the people to have germplasm resources, improve the degree of improved sheep varieties, and increase the output of mutton in Xinjiang. [0003] Gonadotropin Releasing Hormone Receptor (GnRHR) is a high-affinity G protein-coupled receptor located on the surface of pituitary gonadotrophs, which can transduce extracellular ligands and hypothalamic factors. The thalamic gonadotropin releasing hormon...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6888C12Q1/686C12Q2600/124C12Q2600/156
Inventor 刘武军邵勇钢王琼田佳努孜古丽·图尔荪
Owner 刘武军