Drought-resistant and salt-tolerant gene IpNY-B1 as well as encoding protein and application thereof

An ipny-b1, protein technology, applied in the field of plant genetic engineering and molecular biology, can solve problems to be developed and so on

Active Publication Date: 2019-03-22
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 NY-B-encoded protein and the role of the gene of Austenitum have yet to be explored

Method used

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  • Drought-resistant and salt-tolerant gene IpNY-B1 as well as encoding protein and application thereof
  • Drought-resistant and salt-tolerant gene IpNY-B1 as well as encoding protein and application thereof
  • Drought-resistant and salt-tolerant gene IpNY-B1 as well as encoding protein and application thereof

Examples

Experimental program
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Effect test

experiment example 1

[0023] Experimental Example 1: Cloning of the Austeng IpNY-B1 gene and construction of the pYES2-IpNY-B1 vector.

[0024] Specifically, the method comprises the following steps: three steps of total RNA extraction, cDNA library construction and gene cloning. The main steps are summarized as follows:

[0025] (1) Total RNA extraction

[0026] Take about 2g of the mixed sample of thick rattan root, leaves, rattan and leaf buds, add liquid nitrogen to grind thoroughly, and use the MAGEN RNA extraction kit to extract RNA according to the instructions. The obtained RNA was dissolved with 50 μl RNase-free water. The quality and concentration of RNA were determined with a Nanodrop micro-spectrophotometer.

[0027] (2) Construction of cDNA primary library

[0028] Use PromegaM-MLV Reverse Transcriptase Reverse Transcription Kit to synthesize the first strand of cDNA, take about 2 μg RNA, add 0.5 μg Oligo(dT)Primer and RNase-free H2O to a total volume of 15 μL; incubate at 70°C for...

experiment example 2

[0033] Experimental example 2: Overexpression of IpNY-B1 gene in yeast improves the salt tolerance and oxidative stress tolerance of yeast

[0034] (1) Yeast strain transformed with pYES2-IpNY-B1

[0035] The concentration of pYES2-IpNY-B1 recombinant plasmid was adjusted to 1 μg / μL, and the salt-sensitive yeast mutant strain AXT3 and the corresponding wild-type yeast strain (W303) were respectively transformed by lithium acetate transformation method. In addition, using the same method to convert to H 2 o 2 Sensitive yeast mutants skn7Δ and yap1Δ and corresponding wild-type yeast strains (WT). At the same time, the yeast expression vector empty vector pYES2 was used as a control to transform the above yeast strains respectively.

[0036] (2) Overexpression of IpNY-B1 in yeast salt-sensitive mutant strain AXT3 and wild-type strain W303 increased the resistance of transgenic yeast

[0037] Pick the single clones of yeast strain AXT3 transformed with the IpNY-B1 gene overexp...

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Abstract

The invention relates to a drought-resistant and salt-tolerant gene IpNY-B1 as well as an encoding protein and an application thereof. The gene IpNY-B1 is obtained by cloning from Ipomoea pes-caprae as an important wild plant in sand beach, and drought resistance and salt tolerance of plants can be improved by transferring the gene into cells, tissue or individuals of host plants; resistance of engineered strains to high salinity and oxidative stress can be improved by transferring the gene into saccharomyces cerevisiae; the gene has remarkable application prospects in the field of improvementof resistance of the plants and the engineered strains.

Description

technical field [0001] The invention belongs to the technical fields of plant genetic engineering and molecular biology, and relates to a drought-resistant and salt-tolerant gene IpNY-B1 and its encoded protein and application. Background technique [0002] Unfavorable factors such as abiotic stresses (such as drought, salt, heavy metals, and extreme temperatures) have severely jeopardized the survival of many plants, causing hundreds of billions of dollars in crop losses worldwide each year. In addition, the emergence of problems such as population growth, reduction of arable land, and agricultural production reduction has made it urgent to increase crop production. Understanding the formation mechanism of plant stress resistance, mining important stress resistance gene resources, and cultivating new crop varieties with stress resistance have important application value and strategic significance. my country is one of the countries with the most serious soil salinization. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82C12N1/19A01H5/00A01H6/46A01H6/54A01H6/20C12R1/865
CPCC07K14/415C12N15/8273
Inventor 罗鸣张美杨超徐颖超储刘甜周晓晨
Owner SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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