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Casuarina equisetifolia gene CeDREB1 and application thereof

A casuarina and genetic technology, applied in the field of genetic engineering, to achieve wide application value, improve tolerance, and improve drought resistance

Active Publication Date: 2020-08-07
SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the important anti-stress genes of casuarina have yet to be excavated and developed

Method used

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  • Casuarina equisetifolia gene CeDREB1 and application thereof
  • Casuarina equisetifolia gene CeDREB1 and application thereof
  • Casuarina equisetifolia gene CeDREB1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1 Casuarina drought-resistant gene CeDREB1 and its expressed protein

[0050] In this example, a casuarina drought-resistant gene CeDREB1 has a cDNA sequence as shown in SEQ ID NO.1, or a sequence that is completely complementary to SEQ ID NO.1, or an encoded amino acid sequence as shown in SEQ ID NO.2 the sequence of.

[0051] The amino acid sequence of the expressed protein of the Casuarina drought-resistant gene CeDREB1 is shown in SEQ ID NO.2.

[0052] The cDNA of the above-mentioned casuarina drought-resistant gene CeDREB1 was obtained by the following method:

[0053] 1. Extraction of Casuarina RNA

[0054] Casuarina RNA was extracted using the Plant RNA Kit of Beijing Huayueyang Biotechnology Co., Ltd., an ultra-fast plant RNA extraction kit.

[0055] (1) Weigh 100 mg of fresh roots and leaves of Casuarina elata, put them into a mortar quickly, add liquid nitrogen and grind to crush.

[0056] (2) Put the ground plant tissue into a centrifuge tube and ...

Embodiment 2

[0097] Example 2 A recombinant expression vector pYES-DEST52-CeDREB1 of the CeDREB1 gene

[0098] A recombinant expression vector of the CeDREB1 gene in this embodiment, its structure schematic diagram is as follows figure 2 As shown, it is obtained by inserting the casuarina drought-resistant gene CeDREB1 described in Example 1 into the pYES-DEST52 yeast expression vector, and the specific construction method includes the following steps:

[0099] 1. Obtaining pYES-DEST52 enzyme-cut vector

[0100] Use Thermo Scientific restriction enzymes and buffers

[0101] (1) Prepare pYES-DEST52 digestion reaction solution: 5.8 μL of yeast plasmid, 1 μL of BamHI, 1 μL of EcoRI, 5 μL of K buffer (1×), add distilled water to make up to 50 μL.

[0102] (2) Place the enzyme digestion reaction solution in a 37°C water bath for 3 hours.

[0103] 2. The casuarina CeDREB1 gene was connected to the pYES-DEST52 vector after digestion.

[0104] Use Sangon Biotechnology Co., Ltd. Ready-to-Use Sea...

Embodiment 3

[0136] Example 3 A recombinant expression vector pCAMBIA1300-CeDREB1 of the CeDREB1 gene

[0137] A recombinant expression vector of the CeDREB1 gene in this embodiment, its structure schematic diagram is as follows image 3 As shown, it is obtained by inserting the casuarina drought-resistant gene CeDREB1 described in Example 1 into the pCAMBIA1300 plasmid, specifically comprising the following steps: reacting the DNA solution obtained by recovering the glue in Example 1 and the pCAMBIA1300 vector through the Infusion system, and then the Infusion product Import into competent cells, pick positive single colonies, expand culture, extract plasmids, and send the obtained plasmids to a sequencing company for sequencing. The sequencing is completed by Guangzhou Qingke Biological Company. The plasmid with the correct sequencing result is the recombinant expression vector pCAMBIA1300 of the CeDREB1 gene. -CeDREB1.

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Abstract

The invention provides a casuarina equisetifolia gene CeDREB1 and an application of the casuarina equisetifolia gene CeDREB1. The cDNA sequence of the casuarina equisetifolia drought-resistant gene CeDREB1 is shown as SEQ ID NO.1, or is a sequence which is completely complementarily paired with the SEQ ID NO.1, or is a sequence with a coded amino acid sequence shown as SEQ ID NO.2. Functional research is carried out through gene cloning, and the CeDREB1 gene can improve drought resistance of organisms, can be widely applied to improvement of sexual drought resistance of the organisms, comprises construction of drought-resistant saccharomyces cerevisiae engineering strains, cultivation of drought-resistant plant varieties, improvement of drought resistance of plants and the like, and has awide application value in the fields of microorganisms and agriculture.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a casuarina gene CeDREB1 and its application. Background technique [0002] Plants grow in a fixed location for a long time and are more susceptible to adversity stress. Abiotic stress is a harmful factor affecting the growth and productivity of plants and crops, mainly including drought stress, water stress, salt stress and heavy metal stress. Drought stress and salt stress are common types of stress. Water loss will cause changes in plant cell osmotic pressure and damage to plasma membrane peroxidation cell structure, which will affect plant physiological activities and inhibit plant growth. Global climate change leads to frequent occurrence of extreme weather, and crops will face the potential threat of environmental stress. Increasing crop production is particularly important for ensuring food security. The development of crop varieties with strong s...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415C12N15/82C12N15/81A01H5/00A01H6/46A01H6/54
CPCC07K14/415C12N15/81C12N15/8273
Inventor 张世柯刘楠罗鸣周雯雯吴海鹏
Owner SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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