A kind of casuarina gene cesc20 and its application

A casuarina, genetic technology, applied in the field of genetic engineering

Active Publication Date: 2020-12-25
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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  • A kind of casuarina gene cesc20 and its application
  • A kind of casuarina gene cesc20 and its application
  • A kind of casuarina gene cesc20 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1 Casuarina drought-resistant gene CeSC20 and its expressed protein

[0049] In this example, a casuarina drought-resistant gene CeSC20 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.

[0050] The amino acid sequence of the expressed protein of the Casuarina drought-resistant gene CeSC20 is shown in SEQ ID NO.2.

[0051] The cDNA of the above-mentioned casuarina drought-resistant gene CeSC20 is obtained by the following method:

[0052] (1) Take the fresh roots and leaves of casuarina to extract RNA

[0053] Casuarina RNA was extracted using the Plant RNA Kit of Beijing Huayueyang Biotechnology Co., Ltd., an ultra-fast plant RNA extraction kit.

[0054] 1) Weigh 100 mg of fresh roots and leaves of Casuarina elata, quickly put them into a mortar, add liquid nitrogen and grind to crush.

[0055] 2) Put the ground plant tissue int...

Embodiment 2

[0095] Example 2 A recombinant expression vector pYES-DEST52-CeSC20 of the CeSC20 gene

[0096] The recombinant expression vector of a kind of CeSC20 gene of this embodiment, its structure schematic diagram is as follows figure 2 As shown, it is obtained by inserting the casuarina drought-resistant gene CeSC20 described in Example 1 into the pYES-DEST52 yeast expression vector, and the specific construction method includes the following steps:

[0097] 1. Obtaining pYES-DEST52 enzyme-cut vector

[0098] Use Thermo Scientific restriction enzymes and buffers

[0099] (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.

[0100] (2) Place the enzyme digestion reaction solution in a 37°C water bath for 3 hours.

[0101] 2. The casuarina CeSC20 gene was connected to the pYES-DEST52 vector after digestion.

[0102] Use Sangon Biotechnology Co., Ltd. Ready-to-Us...

Embodiment 3

[0134] Example 3 A recombinant expression vector pCAMBIA1300-CeSC20 of the CeSC20 gene

[0135] The recombinant expression vector of a kind of CeSC20 gene of this embodiment, its structure schematic diagram is as follows image 3 As shown, it is obtained by inserting the casuarina drought-resistant gene CeSC20 described in Example 1 into the pCAMBIA1300 vector, 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 CeSC20 gene. -CeSC20.

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Abstract

The invention provides a casuarina equisetifolia gene CeSC20 and an application of the casuarina equisetifolia gene CeSC20. A cDNA sequence of the casuarina equisetifolia gene CeSC20 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 which codes an amino acid sequence shown as SEQ ID NO.2. Functional research is carried out through gene cloning, and the CeSC20 gene can improve the drought resistance of organisms, can be widely applied to improvement of the drought resistance of the organisms, comprises construction of drought-resistant saccharomyces cerevisiae engineering strains, cultivation of drought-resistant plant varieties, improvement of the drought resistance of plants and the like, and has wide 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 CeSC20 and an application thereof. Background technique [0002] Environmental factors for plant growth, such as heavy metals, drought, high salinity, high temperature and low temperature, etc. will affect the growth and development of plants. Among the many factors affecting plant growth and development, drought and high salinity are the most common types. Drought stress affects the distribution and transportation of plant photosynthetic products, resulting in a decrease in plant productivity. Salt stress breaks the ion balance of plant cells and destroys the balance of production and removal of reactive oxygen species in plant cells. The arid and high-salt regions of the world are huge, and the survival, growth and development of plants and crops are faced with challenges of drought and salt stress. The study of stress-resistant genes ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/82C12N15/81A01H5/00A01H6/46A01H6/54
CPCC12N15/81C12N15/8273
Inventor 罗鸣张世柯刘楠周雯雯
Owner SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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