Transgenic plant and preparation method thereof

A technology of transgenic plants and genes, applied in the field of plant genetic engineering, can solve the problems of undiscovered anti-oxidative stress, etc., and achieve the effect of accurate preparation and simple operation

Active Publication Date: 2019-11-12
JIANGHAN UNIVERSITY
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, it has been found that overexpressing the CBF1 gene and CBF3 gene of Brachypodium CBF / DREB can improve the ability of plants to resist cold and drought, but no gene has been found that can Improve plant resistance to oxidative stress

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
  • Transgenic plant and preparation method thereof
  • Transgenic plant and preparation method thereof
  • Transgenic plant and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0038] The embodiment of the present invention provides a preparation method for preparing the above-mentioned transgenic plants, the preparation method comprising:

[0039] Extract the RNA of Brachypodium distachyon;

[0040] Reverse transcription of RNA into cDNA;

[0041] Using cDNA as a template, using the first forward primer and the first reverse primer to carry out PCR amplification to obtain the amplified product, the sequence of the first forward primer is shown in SEQ ID NO: 2 in the sequence table, and the first reverse primer The sequence of the primer is shown in SEQID NO:3 in the sequence listing;

[0042] connecting the amplified product to a carrier to obtain a ligated product;

[0043] Transfer the ligated product into competent cells to obtain the transformed product;

[0044] Positive single clones were screened out from the transformation products to obtain recombinant plasmids;

[0045] Insert the DREB1-like gene on the recombinant plasmid into the bac...

Embodiment 1

[0064] An embodiment of the present invention provides a transgenic plant, the transgenic plant includes a DREB1-like gene, and the sequence of the DREB1-like gene is shown in SEQ ID NO: 1 in the sequence listing.

[0065] The transgenic plants provided by the embodiments of the present invention include the DREB1-like gene, which can endow the transgenic plants with the characteristics of anti-oxidative stress, and provide good climatic gene resources for plant breeding work, and at the same time utilize the transgene of the DREB1-like gene Studying the molecular mechanism of anti-oxidation in plants can facilitate the study of the function of their homologous genes.

Embodiment 2

[0067] The embodiment of the present invention provides a preparation method for preparing transgenic plants, the preparation method comprising:

[0068] The RNA of Brachypodium distachyon was extracted; at the time of realization, 0.1 g of fresh leaves of Brachypodium distachyon was taken, and the total RNA was extracted by the Trizol method. The source of Brachypodium distachyda is preserved by the laboratory of Jianghan University.

[0069] The RNA was reverse-transcribed into cDNA; at the time of implementation, the cDNA was obtained using a cDNA synthesis kit purchased from TaKaRa Company.

[0070] Using cDNA as a template, use the first forward primer and the first reverse primer to perform PCR (Polymerase Chain Reaction, polymerase chain reaction) amplification to obtain the amplified product pBdDREB1-like, the sequence of the first forward primer is shown in the sequence table Shown in SEQ ID NO: 2, the sequence of the first reverse primer is shown in SEQ ID NO: 3 in ...

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 discloses a transgenic plant and a preparation method thereof. The transgenic plant comprises a DREB1-like gene shown as SEQ ID NO:1 in a sequence table. The preparation method comprisesthe steps that RNA of brachypodium distachyon is extracted; the RNA is subjected to inverse transcription into cDNA; the cDNA is taken as a template, and a first forward primer and a first reverse primer are utilized for PCR amplification to obtain an amplification product; the amplification product is connected with a carrier to obtain a connected product; the connected product is transferred into a competent cell to obtain a converted product; positive monoclone is screened from the converted product to obtain recombinant plasmid; the DREB1-like gene on the recombinant plasmid is inserted into a skeleton carrier to obtain a recombinant carrier; the recombinant carrier is transferred into agrobacterium to obtain recombinant bacteria; the recombinant bacteria are transferred into a plantto obtain the T0 generation of transgenic plant; and the T0 generation of positive transgenic plant is screened. The transgenic plant comprises the DREB1-like gene, and the DREB1-like gene can give the transgenic plant the characteristic of resisting oxidative stress.

Description

technical field [0001] The invention relates to the field of plant genetic engineering, in particular to a transgenic plant and a preparation method thereof. Background technique [0002] Plants are often stressed by harsh environments such as low temperature, drought and high salinity during their growth. When plants are subjected to these stresses, they generally produce a large amount of reactive oxygen species (Reactive Oxygen Species, ROS). The accumulation of a large amount of reactive oxygen species will cause oxidative stress to plants, which will cause a series of redox reactions and change the antioxidant system. [0003] At present, it has been found that overexpressing the CBF1 gene and CBF3 gene of Brachypodium CBF / DREB can improve the ability of plants to resist cold and drought, but no genes have been found that can improve the ability of plants to resist oxidative stress. Contents of the invention [0004] In order to solve the problems of the prior art, t...

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): C12N15/82A01H5/00A01H6/82
CPCC12N15/8271C12N15/8205
Inventor 陈利红黄钢葛艳柯杰李论
Owner JIANGHAN UNIVERSITY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap