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Novel milchgoat multiparous and monotocous ovary differential expression associated gene, screening method thereof, identification method thereof and functional analysis method thereof

A differential expression and dairy goat technology, applied in the field of molecular genetics, can solve problems such as low heritability and increase lambing rate

Inactive Publication Date: 2011-02-09
NORTHWEST A & F UNIV
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AI Technical Summary

Problems solved by technology

[0004] The multiple lamb trait is a quantitative trait controlled by a micro-effect polygene, with a very low heritability (about 0.23), and it is difficult to rapidly increase the lambing rate in a short period of time using traditional breeding methods

Method used

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  • Novel milchgoat multiparous and monotocous ovary differential expression associated gene, screening method thereof, identification method thereof and functional analysis method thereof
  • Novel milchgoat multiparous and monotocous ovary differential expression associated gene, screening method thereof, identification method thereof and functional analysis method thereof
  • Novel milchgoat multiparous and monotocous ovary differential expression associated gene, screening method thereof, identification method thereof and functional analysis method thereof

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Experimental program
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Embodiment Construction

[0034] A. Extraction of total RNA from ovarian tissue

[0035](1) Take the ovarian tissue sample out of the liquid nitrogen tank, place it in a mortar pre-cooled with liquid nitrogen, add a small amount of liquid nitrogen and quickly grind it (the mortar should be kept in the presence of liquid nitrogen during the grinding process) until the tissue is When ground into a fine powder, transfer it to a 1.5mL centrifuge tube treated with 0.1% DEPC water and sterilized by high pressure steam (wash the mortar and dry it, burn it with anhydrous ethanol for 5-10min, use after cooling).

[0036] (2) Immediately add Trizol A+ (30-50 mg tissue sample / mL Trizol), invert and mix, and leave at room temperature for 5 minutes to fully lyse the tissue sample. Also, the tissue volume should not exceed 10% of the Trizol volume, otherwise it will lead to DNA contamination.

[0037] (3) Add chloroform according to 0.2 mL chloroform / mL Trizol, shake vigorously for 15 seconds, and leave at room ...

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Abstract

The invention discloses a novel milchgoat multiparous and monotocous ovary differential expression gene, and a screening method thereof, an identification method thereof and a functional analysis method thereof. Xinong Saanen milchgoats are taken as experimental materials, the forward and reverse subtracted cDNA library of ovarian tissue in a milchgoat multiparous (3 goats) and monotocous anestrus is constructed by using suppression subtractive hybridization technology, differential expression gene fragments are screened, the differential expression gene is verified by real-time quantitative polymerase chain reaction (PCR) and the gene structure and potential function are analyzed by bioinformatics.

Description

technical field [0001] The invention belongs to the field of molecular genetics, and specifically relates to a new gene differentially expressed in the ovaries of multiple (three lambs) and singleton dairy goats and its screening, identification and functional analysis methods. Six new genes that are closely related to the growth and development of follicles have been found Gene fragments, which have not been reported in goats. In addition, the two new gene fragments do not match any sequences in NCBI. Background technique [0002] It is well known that "a cup of milk strengthens a nation", however, due to genetic differences between Asians and Westerners, 70% of Chinese people who eat milk suffer from lactose intolerance (He Wei, 2006), but those who eat goat milk do not Lactose intolerance, therefore, goat milk has become a consumption fever in China, and the low lambing rate of dairy goats has become a bottleneck problem restricting the development of the dairy goat indus...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/12C12N15/10
Inventor 曹斌云安小鹏侯金星王建刚宋宇轩杨明明
Owner NORTHWEST A & F UNIV
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