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Target gene binding sequence of miR-25, recombinant vector, transformant and application thereof

A mir-25 and combined sequence technology, applied in the field of genetic engineering, can solve unreported problems and achieve the effect of promoting fat deposition, large-throughput screening and evaluation platforms and means

Pending Publication Date: 2019-01-18
INST OF ANIMAL SCI & VETERINARY HUBEI ACADEMY OF AGRI SCI
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  • Target gene binding sequence of miR-25, recombinant vector, transformant and application thereof
  • Target gene binding sequence of miR-25, recombinant vector, transformant and application thereof
  • Target gene binding sequence of miR-25, recombinant vector, transformant and application thereof

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[0022] A target gene binding sequence of miR-25 provided by the present invention, the nucleotide sequence of the target gene binding sequence is shown in SEQ ID NO.1, and the target gene binding sequence is located at the 3'UTR part of the bovine ACOX1 gene , wherein the nucleotide sequence of the 3'UTR part of the bovine ACOX1 gene is shown in SEQ ID NO.2.

[0023] In order to verify the binding effect of the target gene binding sequence of miR-25, we constructed the recombinant pmirGLO-3'UTR wild-type vector containing the binding sequence, the deletion vector lacking the binding sequence, and the target gene binding to miR-25 A mutant carrier with mutated sequences, and the regulation of the binding sequence on the 3'UTR of the bovine ACOX1 gene was observed using the luciferase reporter gene system. The specific steps are as follows:

[0024] 1. Bioinformatics analysis and amplification of 3'UTR of bovine ACOX1 gene

[0025] 1. ACOX1 gene 3'UTR bioinformatics analysis

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Abstract

The invention belongs to the technical field of genetic engineering and provides a target gene binding sequence of miR-25, a recombinant vector, a transformant and application thereof. The nucleotidesequence of the target gene binding sequence is shown in SEQ ID NO. 1. The recombinant vector comprises the miR-25. The transformant comprises the recombinant vector. The invention defines miR-25 target that 3 'UTR sequence of the ACOX1 gene, and the miR-25 can bind to the binding sequence of 3 'UTR of ACOX1 gene and inhibit the expression of ACOX1 significantly. Not only can it clarify miR-25 onthe post-transcriptional level of ACOX1 gene, which is helpful for the development of inhibitors of ACOX1 gene and the further study of miR-25 in livestock meat quality traits laid the foundation forregulation.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a target gene binding sequence of miR-25, a recombinant vector, a transformant and applications thereof. Background technique [0002] MicroRNAs (miRNAs) are a class of endogenous small single-stranded non-coding RNA molecules with a length of about 22 nt. The functions of different miRNAs in biological systems are determined. They usually bind to the 3'UTR of mRNAs, causing mRNAs to be Degrade or inhibit the translation of mRNA, thereby negatively regulating gene expression (Bartel 2004; Carthew and Sontheimer 2009; Malan-Muller et al., 2013). Currently, thousands of miRNAs have been identified to participate in many biological processes, such as cell proliferation, differentiation, development, and metabolism (Ning et al., 2014). [0003] As a member of the miR-106b-25 cluster, miR-25 is involved in many biological processes. Our previous research results showed t...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12N15/85C12N5/10C12N15/53C12Q1/02
CPCC12N9/0069C12N15/11C12N15/85C12N2310/10C12Y113/12007G01N33/5008
Inventor 张凤陈明新陶虎熊琪李晓锋索效军张年杨前平刘洋张鹤山田宏熊军波
Owner INST OF ANIMAL SCI & VETERINARY HUBEI ACADEMY OF AGRI SCI
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