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Barbadosnut cold-induced transcription factor, its encoding gene and uses

A transcription factor, cold-induced technology, applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve the problems of plant stress resistance, plant stress resistance has not been fully improved, single-function genes, etc., to achieve high practicality The effect of application value, improvement of stress resistance, and broad application prospects

Inactive Publication Date: 2006-07-12
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the complexity of plant stress resistance traits, it is very difficult to improve plant stress resistance using traditional breeding methods. With the development of molecular biology, improving plant stress resistance through genetic processes has opened up a new way of plant stress resistance breeding. , but the isolation of high-efficiency stress-resistant genes has become the main factor limiting plant stress-resistant genetic engineering
In the past, single functional genes were mainly cloned and applied, such as betaine synthase gene and proline synthase gene, etc. Although some effects have been achieved, the stress resistance of plants has not been fully improved

Method used

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  • Barbadosnut cold-induced transcription factor, its encoding gene and uses
  • Barbadosnut cold-induced transcription factor, its encoding gene and uses
  • Barbadosnut cold-induced transcription factor, its encoding gene and uses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1, the acquisition of the full sequence of Jatropha curcas cold-induced transcription factor gene JcDREB cDNA

[0044]The acquisition of the full-length cDNA sequence of Jatropha curcas cold-induced transcription factor gene JcDREB comprises the following steps:

[0045] 1. Cloning of Jatropha curcas cold-induced transcription factor gene JcDREB 3' end sequence

[0046] 1. Plant material processing and total RNA extraction

[0047] Firstly, the Jatropha curcas seedlings were used as materials, and the total RNA was extracted after low-temperature treatment at 4°C for 12 hours, and then detected by 1% agarose gel electrophoresis. The electrophoresis bands are 28s RNA and 18s RNA respectively from top to bottom, indicating that the total RNA with higher purity and integrity has been obtained.

[0048] 2. Cloning of Jatropha curcas cold-induced transcription factor gene JcDREB 3' end sequence

[0049] Compare the amino acid residue sequences of the published ...

Embodiment 2

[0065] Example 2, Bioinformatics analysis of JcDREB and its encoded protein

[0066] 1. Sequence analysis of JcDREB gene and prediction of structure and function of its encoded protein

[0067] The full-length cDNA sequence of JcDREB obtained in Example 1 was used for bioinformatics analysis using DNAMAN software, and its structural diagram is shown in Figure 5 (single line indicates 3'-UTR; box indicates open reading frame, containing 2 functional motifs , the black box represents the AP2 domain, and the hatched line on the right represents the acidic activation region), the total length of the sequence is 768bp, from the 1st to the 768th base of the 5' end is ORF, and the encoding consists of 256 amino acid residues Protein, its estimated molecular weight is 31.731kDa, and its isoelectric point pI value is 9.37. Using the SMART tool to predict the function of the deduced amino acid residue sequence, the result is that the protein contains a typical EREBP / AP2 domain, that is...

Embodiment 3

[0072] Embodiment 3, the structural analysis of JcDREB genomic DNA

[0073] Extract the genomic DNA of the cold-treated Jatropha curcas seedlings in Example 1 and use it as a template, and under the guidance of primers JcDREBW-1 and JcDREBW-2, carry out PCR amplification analysis of the structure of the JcDREB gene. After the reaction, the The PCR products were detected by 1% agarose gel electrophoresis, and the detection results are shown in Figure 9 (lane M is the molecular weight standard Marker III, and lane 1 is the PCR amplification product). As a result, a specific band with a length of about 800 bp was amplified by PCR. Using the cDNA of JcDREB as a template, carry out PCR amplification with the above-mentioned identical primers. After the reaction, the PCR product is detected by 1% agarose gel electrophoresis, and the detection results are as shown in Figure 9 (swimming lane M is the molecular weight standard Marker III , Lane 2 is the PCR amplification product), and ...

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Abstract

The invention discloses a Jatropha Curcas transcription factor, its encoding gene and use in cultivating plants with increased resistance. The transcription factor is the protein having one of the following amino acid residue sequences: (1) SEQ ID No.1 in the sequence table, (2) protein with plants resistance regulation of transcription activation function obtained through subjecting SEQ ID No.1 amino acid sequences in the sequence table to 1-10 amino acid sequences substitution, deletion or addition.

Description

technical field [0001] The present invention relates to a stress-related transcription factor and its coding gene and application in plants, in particular to a Jatropha cold-induced transcription factor and its coding gene derived from the EREBP / AP2 family and its improvement in stress resistance in cultivation application in plants. Background technique [0002] Unfavorable natural environments such as low temperature, high temperature, drought, and salinity act on plants, which will cause a series of physiological metabolic reactions in plants, manifested as reversible inhibition of metabolism and growth, and even cause irreversible damage in severe cases, resulting in the entire Plants die. Among various stresses, drought, salinity, and cold (freezing) damage have particularly prominent effects on plants, and are the most important environmental factors affecting plant growth and crop yield. Using genetic engineering as soon as possible and effectively to improve crop d...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N1/21A01H4/00A01H5/00
Inventor 沈世华唐明娟刘杰
Owner INST OF BOTANY CHINESE ACAD OF SCI
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