Five microRNA genes adopting populus cellulose synthase gene as target gene and applications thereof

A technology of cellulose synthase and gene, applied in DNA/RNA fragments, applications, genetic engineering, etc., can solve problems such as functions and regulatory mechanisms that have not been elucidated clearly

Inactive Publication Date: 2015-01-21
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Many studies have shown that miRNAs play an important regulatory role in the process of wood formation. Although multiple miRNAs have been discovered and identified in poplar, their exact functions and regulatory mechanisms have not yet been elucidated, and are still worthy of further study.

Method used

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  • Five microRNA genes adopting populus cellulose synthase gene as target gene and applications thereof
  • Five microRNA genes adopting populus cellulose synthase gene as target gene and applications thereof
  • Five microRNA genes adopting populus cellulose synthase gene as target gene and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 miRNA acquisition

[0047] 1. RNA extraction

[0048] The upper and lower parts of the side branches of the perennial Populus tomentosa and the xylem of the trunk of the same tree were selected, and the materials were collected during the vigorous growth period (July), and quickly put into liquid nitrogen, and RNA was extracted according to the Olebo RNA kit.

[0049] 2. Acquisition and screening of small RNA (small RNA) sequences

[0050] 18-26nt small RNA was recovered with 20% PAGE gel. Then Shanghai Bohao Biotechnology Co., Ltd. completed library construction, 3' and 5' adapter construction and sequencing.

[0051] 3. Identification of miRNA

[0052] Use the BLAST program (NCBI, www.ncbi.nlm.nih.gov) to map the small RNA Signature sequence to the poplar chromosome (JGI annotation information, http: / / genome.jgi.doe.gov / programs / plants / index. jsf), to remove sequences that do not have a match on the genome.

[0053] Compare the miRBase (v18) database of...

Embodiment 2

[0067] Example 2 detection of miRNA expression abundance

[0068] Through high-throughput sequencing, the expression abundance of miRNAs of SEQ ID NO: 1 to SEQ ID NO: 5 in poplar was obtained, and the analysis results are shown in Table 3.

[0069] table 3

[0070]

Embodiment 3

[0071] Embodiment 3 target gene analysis

[0072] Use http: / / plantgrn.noble.org / psRNATarget / website, Maximum expectation is set to 3.0, Length for complementarity scoring (hspsize) is set to 20, Target accessibility is set to 25, and Flanking length around target site for target accessibility analysis is set to 17bp (upstream) and 13bp (downstream), Range of central mismatch leading to translational inhibition is set to 9-11nt, upload miRNAs sequences from SEQ ID NO:1 to SEQ ID NO:5, and select Phytozome v7.0 genome information as the source of target genes , and the analysis results are shown in Table 4.

[0073] Table 4

[0074] SEQ ID NO: Target gene ID 1 POPTR_0001s20920.1 1 POPTR_0008s13360.1 1 POPTR_0004s15490.1 1 POPTR_0019s05670.1 1 POPTR_0010s23340.1 1 POPTR_0014s13350.1 1 POPTR_0005s08970.1 1 POPTR_0009s05370.1 1 POPTR_0013s12080.1 1 POPTR_0017s14310.2

[0075] 1 POPTR_0017s1099...

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Abstract

Five microRNA genes adopting a populus cellulose synthase gene as a target gene and applications thereof are provided. The micRNAs have sequences shown as SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4 or SEQ ID NO:5. The miRNAs have specific expression parts in plants and influence synthesis of cellulose in the specific parts of the plants. After precursor genes of the miRNAs, or mature miRNAs are directly put into expression vectors and transformed into the plants, precursors with occipital ring structures are formed in the plants, and is processed and modified to form mature miRNAs, thus inhibiting or influencing gene expression and achieving an objective of gene modification.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to five microRNA genes whose target gene is the poplar cellulose synthase gene and their application. Background technique [0002] MicroRNAs (miRNAs) are a class of endogenous regulatory non-coding RNAs discovered in eukaryotes in recent years, with a length of about 20-25 nucleotides. miRNA has a variety of important regulatory functions in cells, and is an important class of gene expression regulators. It binds to the corresponding region of mRNA through incomplete base complementation, thereby inhibiting protein translation; in plants, it can also It degrades mRNA and acts as a negative regulator of target gene post-transcription. With the gradual deepening of research on miRNA regulation of gene expression, it is helpful to understand the complexity of higher eukaryotic genomes and the complex gene expression regulatory network. [0003] Wood is the most important renewable resou...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/54C12N15/11C12N15/63
Inventor张德强陈金辉王博文
OwnerBEIJING FORESTRY UNIVERSITY