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Application of Arabidopis thaliana transcription factor at3g46080 gene

A transcription factor, technology of Arabidopsis thaliana, applied in the field of genetic engineering, can solve problems such as secondary pollution and ecological hazards, and achieve the effect of enhancing resistance

Pending Publication Date: 2019-10-08
XUZHOU NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the control of crops mainly adopts chemical agents or biological control methods, but these two common methods will cause secondary pollution, or some other ecological hazards

Method used

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  • Application of Arabidopis thaliana transcription factor at3g46080 gene
  • Application of Arabidopis thaliana transcription factor at3g46080 gene
  • Application of Arabidopis thaliana transcription factor at3g46080 gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1 Obtaining of Arabidopsis transcription factor at3g46080 gene sequence and construction of plant expression vector

[0019] Log in to the Arabidopsis information resource database website: http: / / www.arabidopsis.org / servlets / TairObject? type=sequence&id=30040, find the sequence of transcription factor at3g46080 gene. Design PCR amplification primers according to the CDS of the at3g46080 gene,

[0020] at3g46080-F:5'-ATGGTTGCGAGAAGTGAGGAA-3'

[0021] at3g46080-R:5'-TGAAGAAAATCGTTCCCAACTC-3';

[0022] The wild-type Arabidopsis Col-0 was used as material, and the leaves of Arabidopsis thaliana grown for 3 weeks under short-day sunlight were used as materials. Total RNA was extracted by TRIzol method, and cDNA was obtained by reverse transcription. The reaction steps are as follows: Grind the leaves of Arabidopsis thaliana under the condition of liquid nitrogen. After the liquid nitrogen is completely volatilized, divide the sample into a centrifuge tube contain...

Embodiment 2

[0025] The acquisition of embodiment 2 transgenic Arabidopsis plants

[0026] (1) Cultivation of plants:

[0027] Arabidopsis thaliana seeds were surface-sterilized with 75% ethanol for 30 seconds, then rinsed once with sterile water, then surface-sterilized with 1% NaClO for 5-8 minutes, and rinsed three times with sterile water. After surface disinfection is completed, the treated seeds are planted on 1 / 2 MS medium plate. The well-seeded plates were sealed with parafilm and placed in the dark at 4°C. After 2-4 days, the plates were placed in a plant light incubator, and the light conditions were 12h light / 12h dark alternately. Eight days after the seeds germinated, the Arabidopsis seedlings were transferred to nutrient soil, grown under the same light conditions, and watered regularly. When the above-mentioned cultivated Arabidopsis grows to 6-8 weeks, the mature fruit pods are cut off for transformation.

[0028] (2) Arabidopsis transformation:

[0029] Transformation ...

Embodiment 3

[0033] Example 3 Validation of PCR detection of fusion gene at3g46080-flag insertion into genome in transgenic Arabidopsis thaliana.

[0034] (1) Extraction of genomic DNA

[0035] Take 50 mg of normal growing Arabidopsis leaves to be tested and add 400 μL plant genomic DNA extraction solution after grinding. The components of the extract are 200mM Tris-Cl pH7.4, 250mM NaCl, 25mM EDTA, 1% SDS. After vortexing and mixing, centrifuge at high speed for 5 minutes, absorb the supernatant and add an equal volume of isopropanol, and precipitate at room temperature for 30 minutes. Discard the supernatant after high-speed centrifugation for 5 minutes, wash the pellet twice with 70% ethanol, and dissolve the pellet in 20 μL of nuclease-free ddH 2 O.

[0036](2) Utilize the specific primer of PGWB11 carrier to carry out PCR amplification

[0037] PCR was performed using the above-mentioned extracted genomic DNA as a template, and the system included 4 μL of 5* buffer, 1 μL of upstrea...

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Abstract

The invention relates to an application of an Arabidopis thaliana transcription factor at3g46080 gene. By constructing a CDS sequence of the Arabidopsis thaliana at3g46080 gene into the upstream of aFLAG protein label by a Gateway technology, Arabidopsis thaliana is conversed, which affects the disease phenotype of Arabidopsis thaliana, screening and identification are carried out, and finally the transgenic plants can be obtained. The analysis of the screened plants carried out so that the plant has enhanced resistance to Pseudomonas lycoposum by Arabidopis thaliana, the gene has the same function for homologous genes in crops, and thus has important significance in production.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and in particular relates to the application of Arabidopsis transcription factor at3g46080 gene. Background technique [0002] As a large agricultural and populous country, my country has a large demand for food every year. The output of crops directly affects the supply of agricultural production to the market. Although with the application of traditional breeding technology and modern breeding technology, the yield of crops is increasing year by year, but the market demand for crop yield is also relatively large. However, the yield of crops in agricultural production is always reduced by various factors, and there are many factors affecting crop yield, among which crop diseases and insect pests are one of the important factors affecting global agricultural production. At present, the control of crops mainly adopts chemical agents or biological control methods, but these two common methods w...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/29C12N15/62C12N15/82A01H5/00A01H6/20
CPCC07K14/415C12N15/8281C07K2319/43
Inventor 潘巧娜崔北米加利·约翰·洛克许师文
Owner XUZHOU NORMAL UNIVERSITY
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