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A rice sugar-inducible promoter srn1 and its application

A promoter and rice technology, applied in the field of genetic engineering, can solve the problem that the nucleic acid sequence has not been discovered

Active Publication Date: 2022-03-18
CHINA NAT RICE RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the nucleic acid sequence of a highly efficient sucrose-inducible promoter has not been found

Method used

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  • A rice sugar-inducible promoter srn1 and its application
  • A rice sugar-inducible promoter srn1 and its application
  • A rice sugar-inducible promoter srn1 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1: Sequence Analysis of Rice Promoter SRN1

[0031] Using the plant promoter prediction analysis website PlantCARE ( http: / / bioinformatics.psb.ugent.be / webtools / plantcare / ) and New PLACE ( https: / / www.dna.affrc.go.jp / PLACE / ) to analyze and organize the cis-acting elements of the promoter region of the SRN gene 2000bp (ie promoter SRN1), the results are shown in figure 1 and Table 1.

[0032] Table 1 Annotation of sequence information of cis-acting elements in rice promoter SRN1

[0033]

[0034]

[0035] Depend on figure 1 It can be seen from Table 1 that there are 14 cis-acting elements involved in the expression of 8 kinds of responses in the 2000 bp promoter region of the SRN gene (promoter SRN1), and 74% of the cis-acting elements participate in the sugar response and light response processes. Since there is a great correlation between sugar content and light status in plants, the above results indicate that the promoter SRN1 (ie, SRN1 ge...

Embodiment 2

[0036] Example 2: Construction and transformation of rice proSRN1::SRN1-GFP recombinant vector

[0037] (1) Genomic DNA of rice Nip extracted by CTAB method.

[0038] (2) Using rice Nip cDNA as a template for amplifying the coding sequence of the SRN gene, the primer sequences are as follows:

[0039] SRN F: tcaccaaatccctgcaagaaatggtgagcaagggcgaggag; SEQ ID No. 6;

[0040] SRN R: ctgtacatggtagatcttgcgaagatctaccatgtacagctcgt; SEQ ID No. 7.

[0041] (3) According to the 2000 bp sequence upstream of the ATG transcription initiation site of the SRN gene found on the RGAP (http: / / rice.plantbiology.msu.edu / ) website, the promoter SRN1 primer was designed. Use the high-fidelity enzyme KOD FX to amplify the promoter SRN1, and the primer sequences are as follows:

[0042] SRN1pro F 1 : tctctagaactagtggatccatggcgatgacaccgcagct; SEQ ID No.2;

[0043] SRN1pro R 1 : ataagcttgatatcgaattcctagccaccatggtttct; SEQ ID No.3.

[0044] (4) Utilizing the GFP vector as a template for amplifyin...

Embodiment 3

[0048] Example 3: Observation of sugar-induced rice SRN1 gene (promoter SRN1) expression under a fluorescence microscope

[0049] The construction method of pro35s::GFP is as follows:

[0050] (1) According to the 35s promoter sequence found on the NCBI website (http: / / www.ncbi.nlm.nih.gov / nuccore / AF234297 / ), design primers for the 35s promoter. Use the high-fidelity enzyme KOD FX to amplify the promoter 35S, and the primer sequences are as follows:

[0051] 35s pro F: acgaattcgagctcggtacccatggagtcaaagattcaa; SEQ ID No. 10;

[0052] 35s pro R: agtcccccgtgttctctccaaatgaa; SEQ ID No. 11.

[0053] (2) Utilizing the GFP vector as a template for amplifying the coding sequence of the GFP gene. The GFP gene coding sequence was amplified by the high-fidelity enzyme KOD FX, and the primer sequences were as follows:

[0054] GFP F 2 :agtcccccgtgttctctatgacaccgcagcta; SEQ ID No.12;

[0055] GFP R 2 : ctgtacatggtagatcttgcgaagatctaccatgtacagctc; SEQ ID No. 13.

[0056] (3) Finally ...

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Abstract

The invention discloses a rice sugar-induced promoter SRN1 and its application. It belongs to the technical field of genetic engineering. The nucleotide sequence of the rice sugar-inducible promoter SRN1 is shown in SEQ ID No.1. The promoter SRN1 belongs to a tissue-specific promoter induced by sucrose and glucose, and has important application value in the field of plant genetic engineering and the study of the transduction mechanism of sugar signal molecules in organisms. Promoter SRN1 provides a tool for directional expression of genes for important agronomic traits of rice or other food crops and genetic engineering breeding for adversity stress, and can also be used for scientific research to indicate the level of soluble sugar in plants.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, and more specifically relates to a rice sugar-induced promoter SRN1 and its application. Background technique [0002] The important agronomic traits of crops and the regulation of gene expression in response to adversity stress are one of the research focuses of plant genetic engineering, and the promoter, as an important regulatory sequence to regulate gene expression, is a research hotspot in plant genetic engineering. Under the stimulation of certain physical or chemical signals, the inducible promoter can greatly increase or inhibit the transcription level of the driver gene. Inducible promoters often have sequence structures such as enhancers and silencers, and some of these promoters exhibit tissue-specific expression or rhythmic expression. [0003] According to different induction conditions, inducible promoters can be divided into light-inducible promoters, temperature-indu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/11C12N15/82A01H5/00A01H6/46
CPCC07K14/415C12N15/8223C12N15/8238
Inventor 张健李志永童晓红王以锋应杰政
Owner CHINA NAT RICE RES INST