Supercharge Your Innovation With Domain-Expert AI Agents!

A kind of cotton dehydrin protein and its coding gene and application

A dehydrin protein and gene technology, applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve the problems of response mechanism that needs to be further studied and complicated mechanism

Active Publication Date: 2018-03-16
BIOCENTURY TRANSGENE CHINA
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, as far as the current research status is concerned, due to the complexity of the mechanism, the biochemical and physiological response mechanisms of many plants to stress still need to be further studied.

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
  • A kind of cotton dehydrin protein and its coding gene and application
  • A kind of cotton dehydrin protein and its coding gene and application
  • A kind of cotton dehydrin protein and its coding gene and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Cotton SSH library construction under drought stress in embodiment 1:

[0021] The specific method is:

[0022] According to Clontech's PCR-select TM The method shown in the cDNA Subtraction Kit kit manual was used to construct the SSH library (subtractive library) by suppression subtractive hybridization method. During the experiment, the mRNA of the leaves of the drought-treated cotton seedlings was used as the sample (Tester), and the mRNA of the leaves of the untreated cotton seedlings was used as the control (Driver). Specific steps are as follows:

[0023] (1) Test materials:

[0024] Ji Mian 14 (National Cotton Medium-term Bank, acquired by China Cotton Research Institute, uniform number: ZM-30270) was sown on sterilized vermiculite, and cultivated under light at 25°C (photoperiod 16h light / 8h dark), every week Pour 1 / 2MS liquid medium (containing 9.39mM KNO 3 , 0.625mM KH 2 PO 4 , 10.3mM NH 4 NO 3 , 0.75mM MgSO 4 , 1.5mM CaCl 2 , 50 μM KI, 100 μM HO ...

Embodiment 2

[0036] Cloning of embodiment 2 cotton dehydrin gene dehydrin09

[0037] After the redundant DNA was removed from the sequencing results of the positive clone numbered Gh-D222, the sequence was SEQ ID No: 3. Sequence analysis showed that the protein encoded by this sequence was a dehydrin protein. In this paper, the full-length corresponding to SEQ ID No: 3 The coding gene is named Ghdehydrin09, and its corresponding protein is named dehydrin09.

[0038] SEQ ID No: 3

[0039]

[0040] Cloning of the full-length coding gene of Ghdehydrin09

[0041] The obtained Ghdehydrin09 gene fragment (SEQ ID No: 3) already has a stop codon TGA, so it only needs to do 5' RACE to clone its full-length coding gene.

[0042] According to the obtained sequence of SEQ ID No: 3, three specific primers were designed as reverse transcription primers and 3' end-specific primers of 5' RACE.

[0043] Ghdehydrin09 GSP1: SEQ ID NO: 4:

[0044] TATGATTTGT ACTAAAAATG

[0045] Ghdehydrin09 GSP2: SEQ ...

Embodiment 3

[0067] Embodiment 3 Ghdehydrin09 gene plant expression vector construction

[0068] The plant binary expression vector pCAMBIA2300 (purchased from Beijing Dingguo Changsheng Biotechnology Co., Ltd.) was selected as the plant expression vector, and the 35S promoter of the NPTII gene containing double enhancers was replaced with the Pnos promoter to reduce the expression of NPTII protein in plants . The inducible promoter rd29A and the Tnos terminator were selected as the promoter and terminator of the Ghdehydrin09 gene, respectively, and the construction process is shown in Figure 1.

[0069] Primers SEQ ID NO: 9 and SEQ ID NO: 10 were used to amplify Pnos with the plant expression vector PBI 121 (purchased from Beijing Huaxia Ocean Technology Co., Ltd.) as a template, and TaKaRa's PrimeSTAR HS DNA polymerase was used. 50 μl PCR reaction system: 10 μl 5×PS Buffer, 3 μl 2.5 mM dNTP, 1.0 μl PBI121, 1.0 μl PrimeSTAR, 10 μM primers SEQ ID NO: 9 and 2.0 μl each of SEQ ID NO: 10, an...

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

A cotton-sourced dehydrin protein dehydrin09, a coding gene of same, and an application thereof in cultivating a transgenic plant of increased drought tolerance.

Description

technical field [0001] The invention relates to plant protein and its encoding gene and application, in particular to a dehydrin protein dehydrin09 derived from cotton and its encoding gene, as well as its application in cultivating transgenic plants with improved drought tolerance. technical background [0002] Dehydrin is a kind of LEA protein, which widely exists in various tissues and organs of plants and in the later stage of plant embryonic development. Dehydrin is a kind of highly hydrophilic protective protein synthesized by plants under abiotic stresses such as low temperature, drought and high salinity. It has the function of protecting nucleic acid, intracellular protein and membrane structure from damage. Many studies have confirmed that there is a close relationship between the expression and accumulation of plant dehydrin and plant stress resistance under abiotic stress. [0003] Abiotic stresses, such as drought, salinity, extreme temperature, chemical pollut...

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
IPC IPC(8): C07K14/415C12N15/05C12N15/29C12N15/63A01H5/00
CPCC07K14/415C12N15/8273
Inventor 陈文华孙超何云蔚崔洪志
Owner BIOCENTURY TRANSGENE CHINA
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

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

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com