Method for screening activity enhancer by utilizing vector based on random sequence

A technology of random sequences and vectors, applied in the field of bioengineering

Pending Publication Date: 2020-03-17
AGRI GENOMICS INST CHINESE ACADEMY OF AGRI SCI
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  • Application Information

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Problems solved by technology

As we all know, due to the structural characteristics of nucleosomes in chromosomes, these traditional methods cannot achieve sequences shorter than 147bp, and it is almost impossible to identify core enhancers in the genome with existing technologies

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  • Method for screening activity enhancer by utilizing vector based on random sequence
  • Method for screening activity enhancer by utilizing vector based on random sequence
  • Method for screening activity enhancer by utilizing vector based on random sequence

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Embodiment Construction

[0048] The following will clearly and completely describe the technical solutions in the embodiments of the present invention with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only some, not all, embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without creative work, any modifications, equivalent replacements, improvements, etc., shall be included in the protection scope of the present invention Inside.

[0049] Such as figure 1 As shown, the present invention homologously recombines the random synthetic sequence (20-50bp) to the upstream of the mini promoter, if the random sequence has enhancer activity, it will activate the expression of the downstream reporter gene mChery, and enrich the enhancer-positive cells by flow cytometry After screening, the enhancer sequence was obtained by bu...

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Abstract

The invention discloses an enhancer screening expression vector and a method for screening an enhancer by utilizing a random sequence. The enhancer screening expression vector is formed by modifying ashuttle plasmid vector, wherein a core component on the enhancer screening expression carrier comprises a mini promoter and a downstream mcherry reporter gene, and the upstream of the mini promoter comprises a recombination site of a random sequence. The invention provides the enhancer screening expression vector and the method for screening the enhancer by using the random sequence. According tothe method, the enhancer sequence is effectively identified by utilizing the enhancer screening expression vector and a screening library; compared with the prior art, the method has the advantages that the identification accuracy of the enhancer in the genome is greatly improved, meanwhile, the enhancer in a dozens of bp sequences can be effectively identified, and the method has important significance in deeply understanding a gene expression regulation mechanism.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to an enhancer screening expression vector and a method for screening enhancers using random sequences. Background technique [0002] Gene expression in eukaryotes is a delicate, complex and dynamic process, which is complexly regulated by multiple factors such as genomic transcription factors, epigenetic modification and various regulatory elements to perform spatiotemporal specific expression of various genes. Among them, cis-regulatory elements change the high-level structure of chromatin by recruiting different trans-acting factors, thereby initiating or silencing gene expression, which is a hot field of gene regulation research in recent years. Identification has important implications for a deeper understanding of the mechanisms underlying gene expression regulation. [0003] Enhancers are a class of cis-regulatory elements that play an important role in the regulation of gene e...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/63C12N15/65C12Q1/6869
CPCC12N15/63C12N15/65C12Q1/6869C12Q2535/122
Inventor 张玉波朱秀生黄雷李清
Owner AGRI GENOMICS INST CHINESE ACADEMY OF AGRI SCI
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