Specific promoter Os023g37190 of rice root and application thereof

A technology of promoter and rice, applied in the field of genetic engineering

Inactive Publication Date: 2011-08-17
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] There are no related reports on the rice Os02g37190 promoter regulating the root-specific expression and application of exogenous genes

Method used

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  • Specific promoter Os023g37190 of rice root and application thereof
  • Specific promoter Os023g37190 of rice root and application thereof
  • Specific promoter Os023g37190 of rice root and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Example 1. Screening of root-specific expression genes:

[0065] 1. Sample preparation

[0066] Select plump Nipponbare wild-type rice seeds, peel them and rinse them with clean water, and then soak them in clean water, soak the seeds at 37°C until they are white (1-2 days), and change the water sooner or later during the period. Directly direct the germinated seeds into 3L rice total nutrient solution (pH 5.5). The mother liquor formula of rice total nutrient solution is shown in (Table 1). The specific preparation of rice total nutrient solution is as follows, that is, No. I-VI is added to every 4L of culture solution 5ml each of the stock solution, adjust the pH to 5.5. Grow for 10 days in a rice growth room with controlled light and temperature (30°C during the day, 22°C at night, 3000 lux light intensity, 12 hours light time). Sampling the ground and roots separately. In addition, the rice spikelets grown in the normal nutrient solution to the heading and flowering s...

Embodiment 2

[0126] Example 2. Expression analysis of rice root-specific promoter Os02g37190 regulating GUS reporter gene

[0127] 1. Construction of GUS vector fused with rice root-specific promoter

[0128] The Os02g37190 promoter primer was designed according to the rice genome sequence (TIGRE), and the restriction site HindIII (shown in lowercase font), BamHI (shown in lowercase italic font) and protected bases (underlined) were added Shown).

[0129] Os02g37190-3.5Kb promoter primer

[0130] F: 5’ aagcttTGACAATCCGCCGGAGTCGC-3’

[0131] R: 5’- ggatccCTGCATGCATATAGACGACA-3’,

[0132] Using genomic DNA as a template, the OsO2g37190 promoter was amplified with phusion ultra-fidelity DNA polymerase, with a total length of 3500 bp, as shown in SEQ ID NO:1. After electrophoresis recovery, the PCR product recovered by double digestion with HindIII and BamHI was connected to the GUS plus vector digested with HindIII and BamHI ( Image 6 ) To obtain the expression vector Os02g37190 promoter: GUS. Tra...

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PUM

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Abstract

The invention discloses a promoter Os023g37190 for regulating specific expression of exogenous gene in rice root. The promoter is a nucleotide sequence shown as SEQ ID NO:1. The promoter Os023g37190 disclosed by the invention can be used for constructing transgenic plant vectors specially expressed in root. The invention also provides a transgenic plant cell which contains the promoter sequence shown in the invention.

Description

Technical field [0001] The present invention belongs to the field of genetic engineering. Specifically, the present invention relates to the promoter sequence of the rice Os02g37190 gene, that is, the nucleotide sequence between -1 and 3500 bp upstream from the ATG of the gene, and the promoter sequence can be used in rice. The transgenic regulatory system drives the specific expression of target genes in roots. The invention also relates to the application of the sequence in genetic engineering and genetic improvement. Background technique [0002] The successful application of plant genetic engineering technology requires not only a large number of different levels of promoters to be regulated, but also promoters suitable for different plant backgrounds, different organs, tissues, and transgenic types to avoid adverse effects in the transgenic process. For a long time, non-plant endogenous constitutive promoters, derived from cauliflower mosaic virus (cauliflower mosaic virus)...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N5/10C12N15/82
Inventor 吴平李援亚张帆陈明秀吴运荣
Owner ZHEJIANG UNIV
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