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

Arabidopsis long-chain fatty alcohol oxidase gene as well as encoding protein and application thereof

A technology of alcohol oxidase and Arabidopsis thaliana, applied in the field of plant genetic engineering, to achieve the effect of improving plant cold resistance

Active Publication Date: 2015-06-03
杭州禾骑士未来生物科技有限公司
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The discovery of cold stress-induced CBFs or DRE-binding factors in Arabidopsis has given people a more specific and in-depth understanding of cold-regulated genes (COR), but whether the AtFao4b gene plays an important role in plant cold resistance remains to be seen. Research

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
  • Arabidopsis long-chain fatty alcohol oxidase gene as well as encoding protein and application thereof
  • Arabidopsis long-chain fatty alcohol oxidase gene as well as encoding protein and application thereof
  • Arabidopsis long-chain fatty alcohol oxidase gene as well as encoding protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] This example illustrates the cloning of the AtFao4b gene.

[0061] Plant material: Arabidopsis variety "col-0", seedlings 4 weeks old.

[0062] 1. AtFao4b RT-PCR primer design

[0063] According to the AtFao4b sequence published by Cheng Qi et al. in the gene bank (www.ncbi.nlm.nih.gov), primers P1 and P2 for AtFao4b gene amplification were designed. The primer sequences are as follows:

[0064] P1: 5'-agatctgatggaagacgttagaagaagaaacagagg-3 (SEQ ID No: 3)

[0065] P2: 5'-cagatctactatacttttgttttgttttgcagcgagtc-3' (SEQ ID No: 4)

[0066] 2. Cloning of AtFao4b cDNA sequence

[0067] Carry out the cloning of AtFao4b cDNA sequence with following method, concrete process comprises the following steps:

[0068] 1. Arabidopsis RNA extraction

[0069] Using TRIZOL from Invitrogen R The Reagent kit (Cat.No.15596-026) was used to extract the leaf RNA of the above-mentioned salicylic acid-induced Arabidopsis variety "col-0" according to the kit instructions. The specific pro...

Embodiment 2

[0097] This example illustrates the construction of AtFao4b plant expression vector pAtFao4b.

[0098] The expression cassette of the AtFao4b gene constructed in this example includes the following gene expression regulatory elements: 35S promoter at the 5' end, AtFao4b and NOS terminator at the 3' end. The specific process includes the following steps:

[0099] The construction process of AtFao4b plant expression vector pAtFao4b is as follows: Figure 5 As shown, the specific method is: the plasmid vector TAtFao4b is digested with the restriction endonuclease BglII, the 2247bp AtFao4b target fragment is recovered, and it is connected with the plant vector pCambia1302 digested by the same enzyme to obtain the AtFao4b plant expression vector, Named pAtFao4b. It was identified by restriction endonuclease BglII and BamH I respectively, and the results of enzyme digestion and identification were as follows: Figure 6 As shown, the fragment size of the digested product is 2247bp...

Embodiment 3

[0101] This example illustrates the molecular detection of AtFao4b transformed Arabidopsis and transgenic plants.

[0102] The plant expression vector pAtFao4b constructed in Example 2 was transformed into Agrobacterium tumefaciens LBA4404 by freeze-thaw method. The Agrobacterium tumefaciens LBA4404 transformant integrated with pAtFao4b was then transformed into Arabidopsis col-0 by staining method, and 8 AtFao4b transgenic Arabidopsis pure lines were obtained.

[0103] The primer sequences for PCR molecular detection are:

[0104] Detection primer 1N: 5'-GGTGCGTTGGATGAGAATGG-3' (SEQ ID No: 5)

[0105] Detection primer 1C: 5'-CCCGTCGTCCTTGAAGAAGAT-3' (SEQ ID No: 6)

[0106] Detection primer 2N: 5'-ATGGTGAGAGCTGGGAAGCA-3' (SEQ ID No: 7)

[0107] Detection primer 2C: 5'-TTGAAGAAGTCGTGCCGCT-3' (SEQ ID No: 8)

[0108] The identification results using different PCR primers are as follows Figure 7 shown.

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 relates to an arabidopsis long-chain fatty alcohol oxidase gene and an encoding protein amino acid sequence of the gene, and also relates to application of the gene on improvement of plant cold resistance and a method. The arabidopsis long-chain fatty alcohol oxidase gene has new application of the improvement of the plant cold resistance and cold stress treatment on the arabidopsis plant of over-expression AtFao4b is carried out. Compared with a contrast, the over-expression AtFao4b plant resistance is obviously enhanced; because of the research on the cold resistance function of the AtFao4b, the cold resistance reaction can be comprehensively known, and the gene has an important meaning and value.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, relates to a plant gene and its coded protein and its application, in particular to a long-chain fatty alcohol oxidase gene from Arabidopsis thaliana and its coded protein and its new application in improving plant cold resistance. Background technique [0002] Only one-third of the land on Earth can maintain a temperature above 0°C, and 42% of the land will experience low temperatures below -20°C. In these regions, plants need special mechanisms to adapt to low temperatures. Generally, cold stress can be divided into chilling injury (0-15°C) and freezing injury (<0°C) according to the temperature. Plants growing in temperate zones, such as Arabidopsis, spinach, etc., have different degrees of tolerance to chilling damage, and they can be placed in a low-temperature environment, and their tolerance to freezing damage can be improved after low-temperature domestication, while plants gr...

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): C12N15/53C12N9/04C12N15/82A01H5/00
Inventor 程奇孙文丽
Owner 杭州禾骑士未来生物科技有限公司
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