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Goat miR-27a (micro Ribonucleic Acid-27a) site-specific modification system and application thereof

A mir-27a, fixed-point modification technology, applied in the goat miR-27a fixed-point modification system and its application field, can solve the problems of low heritability, restricting the economic benefits of the sheep breeding industry, and slow breeding progress

Active Publication Date: 2017-10-24
湖北名羊农业科技发展有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Reproductive traits are quantitative traits, which have defects such as low heritability, long selection cycle, and slow breeding progress, which have become the main reasons for restricting the economic benefits of the sheep breeding industry

Method used

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  • Goat miR-27a (micro Ribonucleic Acid-27a) site-specific modification system and application thereof
  • Goat miR-27a (micro Ribonucleic Acid-27a) site-specific modification system and application thereof
  • Goat miR-27a (micro Ribonucleic Acid-27a) site-specific modification system and application thereof

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Experimental program
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Effect test

Embodiment 1

[0048] 1. Analysis and amplification of goat miR-27a promoter region

[0049] 1.1 Extraction and detection of goat tissue DNA

[0050] The cell / tissue genomic DNA extraction kit (spin column type) provided by Beijing Biotech Biotechnology Co., Ltd. was used to extract the whole genome DNA of goat follicle tissue, and the extracted DNA was stored at -20°C for future use.

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Abstract

The invention belongs to the technical field of animal gene engineering, and particularly relates to a goat miR-27a (micro Ribonucleic Acid-27a) site-specific modification system and application thereof. The goat miR-27a site-specific modification system constructed by the invention can be used for effectively identifying and modifying a binding site of a transcription factor CP2 in a goat miR-27a promoter region. The binding site of the miR-27a promoter of the goat miR-27a site-specific modification system has a nucleotide sequence as shown in SEQ ID NO: 1 in a sequence table. The goat miR-27a site-specific modification system is used for analyzing the binding site of the transcription factor CP2 in the sequence of the goat miR-27a promoter region; a wild dual-luciferase reporter vector pGL3-W of the binding site is constructed; afterwards, the base mutation modification is carried out on the binding site; a mutant dual-luciferase reporter vector pGL3-M is constructed and obtained. The two dual-luciferase reporter vectors constructed by the goat miR-27a site-specific modification system can be used for researching the regulation and control mechanism of the follicular development of a goat, and have important significance for breeding a goat variety with a high-production lamb quantity.

Description

technical field [0001] The invention belongs to the technical field of animal genetic engineering, and specifically relates to a goat miR-27a site-directed modification system and its application, in particular to the editing of the binding site sequence between some specific functional transcription factors and the target gene promoter, for example, in this According to the invention, the binding site of transcription factor CP2 in the miR-27a promoter region is modified, thereby changing the binding ability of CP2 and miR-27a promoter. Background technique [0002] MicroRNA (miRNA) is a single-stranded non-coding RNA encoded by an endogenous gene, generally about 16-29 nucleotides (nucleotide, nt) in length, and regulates its post-transcriptional function by complementary pairing with the 3'UTR of the target gene mRNA. Expression level of (Gu, 2009.Gu S, Jin L, Zhang F, Sarnow P, Kay MA. Biological basis for restriction of microRNA targets to the 3'untranslated region in m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/63C12N15/66C12N15/11
CPCC12N15/11C12N15/63C12N15/66
Inventor 陶虎陈明新熊琪李晓锋索效军张年杨前平刘洋田宏张鹤山熊军波
Owner 湖北名羊农业科技发展有限公司
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