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Method for identifying transcription regulation relationship of plant endogenous siRNA

An identification method and transcriptional regulation technology, applied in the field of molecular genetics, can solve problems such as low efficiency, no analysis method, and inability to perform high-throughput analysis, and achieve the effect of overcoming cumbersome, important theoretical significance and practical value

Active Publication Date: 2020-10-23
BEIJING FORESTRY UNIVERSITY
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Problems solved by technology

At present, the research on the function of siRNA is mainly carried out through experimental methods such as genetic transformation, but the efficiency is too low to carry out high-throughput analysis, and the research on identifying the transcriptional regulation target genes of plant endogenous siRNA by high-throughput sequencing technology has so far been limited. No valid analysis method found

Method used

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  • Method for identifying transcription regulation relationship of plant endogenous siRNA
  • Method for identifying transcription regulation relationship of plant endogenous siRNA
  • Method for identifying transcription regulation relationship of plant endogenous siRNA

Examples

Experimental program
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Embodiment 1

[0046] Experimental materials: Populus tomentosa annual clone '1316' was obtained from the National Populus tomentosa Germplasm Bank;

[0047] The specific operation steps are as follows:

[0048] Select individual leaves of Populus tomentosa for RNA extraction for small RNA sequencing, perform quality control on the original reads obtained by sequencing, and remove adapter sequences and low-quality sequences (including ambiguous base N, sequences with base quality less than 10 and length less than 18nt ) to get clean reads. CLC genomics_workbench 5.5 software was used to align the preprocessed sequences with the Sanger miRBase database (19.0): base mismatches were not allowed during the alignment process. In addition, it will be compared with the data in other non-coding databases ncRNA, piRNA, and Rfam: 2 base mismatches with the target sequence are allowed, and 2 bases are allowed to be shortened or extended from both ends of the target sequence. In the clean reads, the s...

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Abstract

The invention provides a method for identifying the transcription regulation relationship of plant endogenous siRNA, and relates to the technical field of molecular genetics. According to the identification method disclosed by the invention, target genes of plant endogenous siRNA are mined in batches by utilizing a miRNA sequencing and degradation group sequencing data system, so that important information is provided for functional research of the small interfering RNA. According to the identification method disclosed by the invention, a second-generation high-throughput sequencing technologyis fully utilized, high-throughput screening can be carried out on the siRNA and the target genes thereof, and the complexity of a genetic transformation means is overcome; and the method can identify the target genes for siRNA transcriptional regulation more accurately, and has important theoretical significance and practical value for functional research of the siRNA.

Description

technical field [0001] The invention belongs to the technical field of molecular genetics, and in particular relates to an identification method for plant endogenous siRNA transcription regulation relationship. Background technique [0002] Gene expression in eukaryotes is affected by many factors, among which small interfering RNA (small interfering RNA, siRNA) is an important regulator affecting gene expression and plays an important role at the post-transcriptional level. siRNA stimulates the complementary target mRNA to be silenced through the RNA interference (RNAi) mechanism. siRNA is cut into 21-23bp double-stranded RNA by RNase III in the cell from double-stranded RNA (double strand RNA, dsRNA), and siRNA enters the cell and integrates with other proteins to form a silencing complex (RISC). Directed targeted cleavage of mRNA molecules encoding target genes to silence certain gene expression. Compared with miRNA, siRNA usually works by cleaving mRNA before translati...

Claims

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Application Information

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IPC IPC(8): C12Q1/6869G16B30/10
CPCC12Q1/6869G16B30/10C12Q2535/122
Inventor 张德强轩安然宋跃朋
Owner BEIJING FORESTRY UNIVERSITY
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