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An active larch enhancer, its acquisition and identification method

An acquisition method, larch technology, applied in chemical instruments and methods, biochemical equipment and methods, microbe determination/inspection, etc., to achieve rapid identification methods and promote transcription

Active Publication Date: 2022-06-14
YANGZHOU UNIV
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  • Abstract
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  • Claims
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AI Technical Summary

Problems solved by technology

The enhancer sequence is not conserved, and the direction is not fixed, which brings certain difficulties to the enhancer prediction work

Method used

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  • An active larch enhancer, its acquisition and identification method

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Embodiment Construction

[0020] The enhancer acquisition and identification method of this embodiment is as follows:

[0021] 1. How to obtain LkENH1

[0022] (1) Larch RNA-seq: Transcriptome sequencing was performed on the larch needles grown for 30 days and the larch callus induced for 14 days, and the transcripts were assembled using the no-reference genome method, and then combined with the protein database, the expression Transcripts are annotated. ATAC-seq: ATAC-seq sequencing was carried out on larch callus induced and cultured for 14 days to obtain genome open chromatin sequences. Histone ChIP-seq: H3K27acChIP-seq sequencing was performed on larch callus induced and cultured for 14 days to obtain the enriched region sequence of H3K27ac in the genome. In this study, the sequencing data were integrated, and a long fragment (2346bp, SEQ ID NO.2) of larch sequence was obtained by assembly. This sequence includes a transcript with a high expression level and an FPKM value of 0.6046. Because enh...

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Abstract

The invention discloses an active larch enhancer, its acquisition and identification method. The enhancer DNA sequence is as SEQ ID NO.1. Using specific primers to amplify the genomic DNA of Japanese larch as a template to obtain an amplified product of 560 bp. According to the characteristics that enhancers can promote transcription at a long distance, the mini35s promoter alone cannot initiate the expression of the reporter gene GFP. When a functional enhancer is connected to the vector, it can promote the transcription of fluorescent proteins, so that fluorescence can be observed for verification . The larch enhancer sequence provided by the invention is essentially a DNA molecule, 560 bp, which can promote transcription.

Description

technical field [0001] The invention relates to biotechnology, in particular to an active larch enhancer, its acquisition and identification method. Background technique [0002] Larch is a deciduous tree belonging to the family Pinaceae. It has the characteristics of resistance to barrenness and drought, and can grow rapidly in the northern mountains. Therefore, it is one of the main tree species planted in mountainous areas of our country, and it is also the main tree species for papermaking and landscaping. Enhancers are a class of cis-regulatory elements that promote transcription of target genes over long distances. Enhancers act through the promoter to enhance transcription. Enhancer sequences are not conserved and their directions are not fixed, which brings certain difficulties to the work of enhancer prediction. The excavation and identification of enhancers in larch will provide important evidence support for the study of functional genome of larch. Contents ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/10C12N15/82C12N15/65C12Q1/6897
CPCC07K14/415C12N15/10C12N15/8212C12N15/65C12Q1/6897C12Q2531/113
Inventor 张韬刘鹏刘冠卿
Owner YANGZHOU UNIV
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