A kind of plant senescence-specific promoter and application thereof

A plant senescence and promoter technology, applied in the biological field, can solve the problems of resource waste, unfavorable plant healthy development, and plant growth and development poisoning.

Inactive Publication Date: 2011-12-07
SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, genes regulated by constitutive promoters are continuously and efficiently expressed in various stages and tissues of plant development, without specificity in time and space, which is not only an unnecessary waste of resources fo...

Method used

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  • A kind of plant senescence-specific promoter and application thereof
  • A kind of plant senescence-specific promoter and application thereof
  • A kind of plant senescence-specific promoter and application thereof

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Experimental program
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Embodiment approach

[0066] The present invention provides a plant senescence-specific promoter, which has the following characteristics: (a) located upstream of the pheophorbide oxidase gene (called AtPaO in Arabidopsis) (preferably located at Pheophorbide oxidase upstream -2800 to the 200th position in the coding region); (b) has the function of responding to plant senescence signals and initiating the specific expression of the target gene; (c) has the necessary sites for initiating transcription and the transcription initiation point; And (d) the base length is 500-3000. As an embodiment of the present invention, the promoter has the nucleotide sequence shown in SEQ ID NO:1.

[0067] Hybridization of polynucleotides is a technique well known to those skilled in the art, and the hybridization properties of a particular pair of nucleic acids indicate their similarity or identity. Therefore, the present invention also relates to hybridization with the aforementioned specified nucleotide sequence...

Embodiment 1

[0109] Embodiment 1, amplification, cloning and sequence analysis of promoter

[0110] Using Arabidopsis genomic DNA as a template for PCR amplification, a DNA amplification product of about 2093bp was obtained, which was connected to pMD 19-T Vector, and the result was correct after sequencing. The full sequence of the promoter is shown in SEQ ID NO:1.

[0111] This sequence is the promoter of the key enzyme AtPaO in the chlorophyll degradation pathway in Arabidopsis. Therefore, the expression of AtPaO under dark induction conditions was detected by conventional Realtime PCR method.

[0112] It was found that under dark induction conditions, the expression level of AtPaO gene was up-regulated with the increase of dark induction days, see figure 1 . Therefore, it can be seen that the promoter of the AtPaO gene is a senescence-specific promoter.

Embodiment 2

[0113] Embodiment 2, the construction of promoter plant expression vector

[0114] First use Xhol I and BamH I to double digest and recover the 2Kb promoter sequence, then use Sal I and BamHI to double digest pBI 101.3 (purchased from Takara), recover the large fragment, connect the recovered two fragments, transform Escherichia coli, and extract The plasmid was digested and identified, and the promoter was correctly connected into the vector pBI101.3 to obtain pBI101.3-Promoter-GUS.

[0115] The recombinant vector pBI101.3-Promoter-GUS was transformed into Agrobacterium C58 by electric shock transformation method, and after PCR amplification, a band of about 2Kb was identified in 1% agarose gel electrophoresis, which proved that pBI101.3-Promoter-GUS The vector has been transformed into Agrobacterium C58.

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Abstract

The invention relates to a plant aging specific promoter and application thereof. In the invention, a promoter capable of responding to plant aging signals and regulating high expression of target gene specificity is separated for the first time. The promoter is very useful for regulating the expression of the target gene in a plant aging process, so as to change the plant characters, state and aging process.

Description

technical field [0001] The invention belongs to the field of biotechnology; more specifically, the invention relates to a plant senescence specific promoter and application thereof. Background technique [0002] Promoter refers to a DNA sequence in a gene that can accurately and effectively initiate gene transcription, usually located upstream of the gene. Promoter is an important element in the regulation of gene expression. It interacts with trans-acting factors (transcription factors) such as RNA polymerase and other protein cofactors to regulate the level of gene transcription and the specificity of its temporal and spatial expression. [0003] The most widely used constitutive promoter is the tobacco mosaic virus (CaMV) 35S promoter. At the same time, people attach great importance to cloning constitutive promoters from plants themselves and have achieved initial success. The promoters of genes such as actin (Actin) and ubiquitin (Ubiquitin) have been cloned. Mariani ...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/63C12N1/15C12N1/19C12N1/21C12N5/10A01H5/00A01H4/00
Inventor 郭房庆刘芳
Owner SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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