Gene promoter for specific expression of plant trichome

A promoter and trichome technology, applied in the field of plant molecular biology and plant genetic engineering, can solve the problem of little similarity of nucleotide sequences, reduce physical space constraints, simplify recovery and purification, and avoid accumulation Effect

Inactive Publication Date: 2005-08-24
SUN YAT SEN UNIV
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  • Abstract
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Problems solved by technology

These promoters can drive the specific expression of the GUS reporter gene in trichomes, but the nucleotide sequences of these promoters have little similarity

Method used

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  • Gene promoter for specific expression of plant trichome
  • Gene promoter for specific expression of plant trichome

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Experimental program
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Embodiment 1

[0020] Adapter-ligation PCR (Adaptor-ligation PCR) method was used for genomic DNA walking, and the promoter sequence of SaPIN2b gene was amplified and cloned from Solanum nigrum. The kit used for genome walking is GenomeWalker from Clontech TM Kit. Specific steps are as follows:

[0021] 1. Extraction of Solanum nigrum Genomic DNA

[0022] Get 5 grams of Solanum nigrum leaves and grind them into powder in liquid nitrogen, then transfer them to a 30ml centrifuge tube, add 15ml of extraction buffer [100mM Tris (pH8.0), 50mM EDTA (pH8.0), 500mM NaCl, 10mM β-mercapto ethanol], and mix well. Then add 1 ml of 20% SDS to the tube, mix thoroughly, and incubate at 65°C for 10 minutes. Then add 5ml of 5M potassium acetate, mix thoroughly and place on ice for 20 minutes. Centrifuge at 25,000×g for 20 minutes, transfer the supernatant to another clean centrifuge tube, add 10ml of isopropanol, mix well and let stand at -20°C for 30 minutes. Centrifuge at 20,000×g for 15 minutes, re...

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Abstract

A genetic promoter with specific expression to vegetative trichome is disclosed. The sequence of the trichome-specific hydratropic proteinase inhibitor (SaPIN2b) gene promoter is cloned. The start site of SaPIN2b transcription is determined. Its plant expression carrier is configured. The test to its transgenic plant shows its specific expression in trichome.

Description

technical field [0001] The present invention relates to the fields of plant molecular biology and plant genetic engineering, in particular to a gene promoter. Background technique [0002] Like other eukaryotes, the expression of plant genes is regulated at multiple levels such as DNA level, transcription level, post-transcription level, translation level and post-translation level. From the effect point of view, regulation at the transcriptional level is the most economical and effective, so transcriptional regulation is the most important way to control plant gene expression. The transcription of genes into mRNA is regulated by a certain region of the gene, and the region that regulates gene transcription before the transcription start site is usually called a promoter. The region proximal to the transcription initiation site constitutes the "core promoter region"; it typically contains the TATA box and CAAT box sequences, as well as the transcription initiation site. Th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/63C12N15/70C12N15/82
Inventor 徐增富刘进邓宇歌黄小乐
Owner SUN YAT SEN UNIV
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