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Promoter of wheat root specific expression expansin gene and application thereof

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

Inactive Publication Date: 2015-09-23
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the cloning and application of the promoter of the expansin gene specifically expressed in wheat root hairs

Method used

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  • Promoter of wheat root specific expression expansin gene and application thereof
  • Promoter of wheat root specific expression expansin gene and application thereof
  • Promoter of wheat root specific expression expansin gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Specific expression of expansin gene in wheat root TaExpansin expression analysis of

[0022] Material

[0023] The seeds of wheat variety 3338 germinated normally, and the roots and leaves of wheat seedlings were collected after one week. At the heading stage of wheat (when half of the leaf sheaths are pulled out from the young ears), the meristems and young ears of the lower node 0-1 cm of the wheat ears were taken and stored in liquid nitrogen.

[0024] Extraction of Wheat Total RNA by Trizol Method

[0025] 1. Put the tissue material into a liquid nitrogen pre-cooled mortar, and fully grind it into powder in liquid nitrogen;

[0026] 2. After the liquid nitrogen evaporates to dryness, immediately transfer to a 2ml centrifuge tube, add about 1ml of Invitrogen’s TRIzol extract for every 100mg of material, shake and mix the sample vigorously to fully lyse the sample, and place it at room temperature for 5 minutes;

[0027] 3. Add 0.2ml of chloroform (chloroform)...

Embodiment 2

[0080] Cloning of Wheat Root-Specific Expressing Expansin Gene Promoter TaREP

[0081] Extraction of Wheat Genomic DNA

[0082] 1. Genomic DNA was extracted from wheat young leaves by CTAB method. The steps are:

[0083] (1) Take about 0.5g of wheat root system and grind it in liquid nitrogen, put it in a 1.5ml centrifuge tube, add 700μl extraction buffer (100mmol / L Tris-HCl, 50mmol / L EDTA-Na 2 , 100mmol / L NaCl, 1.5% CTAB, 2% mercaptoethanol), 65 o Incubate in C water bath for 30-60min, during which time mix by inverting 3-4 times;

[0084] (2) Take out the centrifuge tube, add 700 μl phenol: chloroform: isoamyl alcohol (25:24:1) solution to each tube, and mix for 10 minutes;

[0085] (3) Centrifuge at 10000rpm for 10min;

[0086] (4) Use a pipette tip (cut off the front 1 / 3) to slowly extract the supernatant into another 1.5ml centrifuge tube;

[0087] (5) Add an equal volume of chloroform:isoamyl alcohol (24:1) and repeat the above extraction process. Slowly remove...

Embodiment 3

[0145] Wheat expansin gene promoter expression vector ( pCAM1391Z / TaREXP) build

[0146] According to the promoter TaREXP sequence design containing Hind III and EcoR The upstream and downstream primers of the promoter of I, the primer name and sequence are REXP1: 5′-CCC AAGCTT CCTTCTGGTCCGAATTGGAC-3' ( Hind III),

[0147] REXP2:5ˊ-CG GAATTC TGATAATCTAGCTATCAGTCAG-3'( EcoR I).

[0148] The plasmid containing the promoter sequence was used as a template for PCR amplification. After electrophoresis and recovery, the PCR amplification product was ligated with the pGEM-T easy vector, transformed into Escherichia coli, and the plasmid was extracted. use Hind III and EcoR I carried out double enzyme digestion on the pCAM1391Z vector and the pGEM-T easy vector containing the TaREXP promoter respectively, recovered the large fragment of the pCAM1391Z vector and the small fragment of the TaREXP promoter, and transformed E. coli DH5α competent cells after ligation wi...

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Abstract

The invention relates to a promoter of a wheat root specific expression expansin gene and an application thereof. The invention aims at providing a promoter of specific expression in a monocotyledon root (especially in root hair). The promoter promotes the specific expression of a target gene in the root. The promoter can be fused with salt-resistant, anti-drought and root secretory protein genes to form a root specific expression carrier, and important significance for breeding of the salt-resistant and anti-drought gene engineering of plants and especially wheat as well as obtaining a root specific expression protein product can be realized.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, and relates to a promoter for specifically expressing expansin gene in wheat roots and an application thereof. Background technique [0002] Promoters are important regulatory elements for gene expression. It directly determines the spatiotemporal expression characteristics and expression amount of genes, so isolating suitable promoters is an important research content of genetic engineering breeding. Tissue-specific expression promoters are a special type of inducible promoters. Under the regulation of these promoters, genes are often expressed only in certain organ tissues. In-depth study of these promoters can not only avoid the waste caused by the continuous expression of constitutive expression promoters, but also help to clarify the basic theories of plant morphology, development, and metabolic pathways, which has a wide range of application values. Example Dapena et al...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/82A01H5/00
Inventor 秦余香刘秀芝
Owner UNIV OF JINAN
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