Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Tetragonia tetragonioides L. TtASR gene as well as encoded protein and application thereof

A gene technology, applied in the field of biological genetic engineering

Active Publication Date: 2019-04-05
GUANGZHOU UNIVERSITY +1
View PDF1 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report on the development and utilization of ASR gene resources in Apricot apricot

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Tetragonia tetragonioides L. TtASR gene as well as encoded protein and application thereof
  • Tetragonia tetragonioides L. TtASR gene as well as encoded protein and application thereof
  • Tetragonia tetragonioides L. TtASR gene as well as encoded protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Obtaining the full-length cDNA of the apricot abscisic acid / stress / maturation-inducing protein gene TtASR by monoclonal sequencing of the apricot cDNA yeast expression library

[0037] 1.1 Construction of the full-length cDNA expression library of Apricot apricot

[0038] The construction of the apricot cDNA library mainly refers to the instruction manual of CloneMiner II cDNA Library Construction Kit, using (Invitrogen) technology. Specifically, the method comprises the following steps: extraction of total RNA, separation of mRNA, construction of a primary cDNA library and construction of a secondary cDNA library. The main steps are summarized as follows:

[0039] (1) Total RNA extraction

[0040] Take 2g of the same amount of apricot leaves, leaf buds, vines and young roots, grind them into powder with liquid nitrogen in a pre-cooled mortar, transfer the powder to multiple RNase-free 1.5mL centrifuge tubes, each centrifuge tube Add 1mL Trizol Reagent, ...

Embodiment 2

[0065] Example 2: Overexpression of TtASR gene in yeast improves salt tolerance and tolerance to oxidative stress of yeast

[0066] 2.1 Transformation of yeast strains with TtASR-pYES-DEST 52

[0067] Adjust the concentration of the TtASR-pYES-DEST 52 recombinant plasmid verified by the sequencing results to 0.1 μg / μL, and use the lithium acetate method to transform the wild-type yeast strain W303 and the yeast pair H 2 o 2 Sensitive mutants yap1Δ and skn7Δ and corresponding wild-type yeast strain WT. At the same time, the yeast expression vector empty vector pYES2 was used as a control, and the above yeast strains were transformed respectively.

[0068] 2.2 The expression of TtASR gene in yeast wild-type strain W303 can improve the salt tolerance of transgenic yeast

[0069] Pick the yeast strain W303 to transform the single clone of empty vector pYES2 and TtASR gene overexpression vector TtASR-pYES-DEST52, inoculate in 2ml of yeast limit liquid medium with galactose added...

Embodiment 3

[0074] Example 3: Overexpression of TtASR gene in Escherichia coli improves salt tolerance and high osmotic pressure tolerance of Escherichia coli 3.1 Construction of Escherichia coli recombinant protein expression vector TtASR-pGEX 6p-1

[0075] With the yeast expression vector pYES-DEST 52 recombinant plasmid containing abscisic acid / stress / maturation-inducing protein gene cDNA as a template, with SEQ ID NO.3 (5'-GGGGCCCCTGGGATCCATGGCCGAAAGCCGTGACAG-3') and SEQ ID NO.4 (5' -GGAATTCCGGGGATCCTTAAAAGAAGTGGTGCTTCT-3') was used as a primer, and the full-length cDNA reading frame of the TtASR gene was amplified by PCR using high-fidelity Taq enzyme. For the PCR system used, refer to the instruction manual of PrimeSTAR HSDNA Polymerase with GC Buffer from TaKaRa Company. The amplified DNA fragments were used according to the instructions of Magen HiPure Gel PureDNA Kits. The recovered fragment was used for insertion into the Escherichia coli recombinant protein expression vector p...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides a tetragonia tetragonioides L. TtASR gene as well as an encoded protein and application thereof, relates to the field of molecular biology and provides a tetragonia tetragonioides L. salt stress response-related gene TtASR. An encoded amino acid sequence of the tetragonia tetragonioides L. salt stress response-related gene TtASR is as shown in SEQ ID NO. 1, and a complete nucleotide sequence of cDNA of the tetragonia tetragonioides L. salt stress response-related gene TtASR is as shown in SEQ ID NO.2. According to the invention, an abscisic acid / stress / maturation inducedprotein TtASR encoded by the TtASR gene is related to the improvement of salt and drought resistance of brewer's yeast, escherichia coli and plants; transgenic overexpression vectors of the TtASR gene in the yeast, escherichia coli and plants are built, so that overexpression of the TtASR gene in the yeast, escherichia coli and arabidopsis is achieved, and the resistance of the yeast, escherichiacoli and arabidopsis to salt stress and drought stress can be improved. The gene can be applied to genetic engineering breeding of engineering bacteria and plants in response to high salt and droughtstress, and has great application value.

Description

technical field [0001] The invention belongs to the field of biological genetic engineering, in particular to TtASR (TtASR) in Tetragonia tetragonioides L. A bscisic acid-, s tress-, and r ipening-induced gene) gene and its encoded protein and its application in regulating salt and drought tolerance of organisms. Background technique [0002] Apricot (Tetragonia tetragonioides L.), also known as New Zealand spinach, wild spinach, sea spinach, and amaranth, is an annual or biennial semi-climbing herb of the Apricot family. It is widely distributed in coastal salt-rich areas in subtropical and temperate regions. , is a saline vegetable with economic and ecological value. In addition, apricot has high nutritional value, rich in minerals and multivitamins, rich in nutrition, fresh and tender fleshy stems and leaves, and refreshing taste. The whole plant of apricot can be used as medicine, which has the effects of clearing away heat and detoxifying, and has certain relieving ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/81C12N15/70C12N1/19C12N1/21C07K14/415A01H5/00A01H6/20C12R1/865C12R1/19
CPCC12N15/70C12N15/81C12N15/8273C07K14/415
Inventor 杨礼香叶玉妍陈红锋罗鸣张美
Owner GUANGZHOU UNIVERSITY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com