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

Liriodendron chinensis cellulose synthase gene LcCESA1 as well as expression protein and application thereof

A technology of cellulose synthase and Liriodendron, applied in the field of plant gene technology, can solve the problem of insufficient research on catalytic synthesis products, and achieve the effect of promoting the accumulation and wide application of cellulose

Active Publication Date: 2018-02-27
NANJING FORESTRY UNIV
View PDF0 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the research on cellulose synthase is relatively mature, the research on all catalytic synthesis products of cellulose and cellulose-like enzymes between the two is not enough.

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
  • Liriodendron chinensis cellulose synthase gene LcCESA1 as well as expression protein and application thereof
  • Liriodendron chinensis cellulose synthase gene LcCESA1 as well as expression protein and application thereof
  • Liriodendron chinensis cellulose synthase gene LcCESA1 as well as expression protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 LcCESA1 clones and LcCESA1 Overexpression vector construction

[0039] 1. Primer design: According to the transcriptome data of Liriodendron tali, compare Arabidopsis thaliana CESA1 CDS sequence prediction of genes LcCESA1 Gene sequence and design specific CDS full-length primers suitable for Gibson Assembly.

[0040] LcCESA1 The CDS full-length primers: Primer-F: 5'-GAGAACACGGGGGACTCTAGAATGGAGGCGAATGCTGGAATGGT-3'; Primer-R: 5'-ATAAGGGACTGACCACCCGGGGCAGTTGATGCACATTGGTTTGC-3'.

[0041] 2. Cloning of the target gene: PCR amplification was performed on the cDNA sample of Liriodendron tulipifera using Q5 ultra-fidelity enzyme to obtain the target gene band.

[0042] (1) System: 3 μL ddH 2 O, 5 μL Q5 ® Ultra-fidelity 2×Master Mix, 0.5 μL Primer-F (10 μM), 0.5 μL Primer-R (10 μM), 1 μL cDNA.

[0043] (2) PCR reaction program: 98°C for 30s; 98°C for 10s, 72°C for 30s, 72°C for 2min, 35 cycles;

[0044] 72°C for 2min; 4°C preservation.

[0045] 3. Connect t...

Embodiment 2

[0058] Example 2 LcCESA1 Identification and Screening of Transgenic Arabidopsis

[0059] 1. Arabidopsis culture

[0060] 1) Sterilization of Arabidopsis thaliana seeds: Take an appropriate amount of wild-type Arabidopsis thaliana seeds and put them in a sterilized EP tube, add an appropriate amount of 75% ethanol, shake for 30s, and then suck out; add the same amount of 0.1% mercuric chloride, and sterilize for 2.5min Aspirate out (discard it in the special waste liquid tank); add the same amount of deionized water, aspirate after shaking and washing, and put the seeds in a new EP tube; repeat the water washing four times.

[0061] 2) Cultivation of Arabidopsis thaliana: put the sterilized seeds in the suspension, and evenly spread them into 1 / 2 MS medium with a pipette gun; seal the medium and put it in a 4°C refrigerator for vernalization for 3 days; The vernalized medium was placed in a constant temperature culture room at 23°C for 7 days; when the Arabidopsis seedlings g...

Embodiment 3

[0092] Example 3 LcCESA1 Observation on the Microstructure of Transgenic Arabidopsis Regenerated Plants

[0093] Prepared with Example 2 LcCESA1 Transgenic Arabidopsis T3 seeds were used as plant material, and wild-type Arabidopsis was used as a control. LcCESA1 Comparison of microstructural observations of transgenic Arabidopsis regenerated plants.

[0094] 1. Arabidopsis culture

[0095] 1) Sterilization of Arabidopsis thaliana seeds: Take an appropriate amount of wild-type Arabidopsis thaliana seeds and put them into a sterilized EP tube, add an appropriate amount of 75% ethanol, shake and sterilize for 30s, and then suck out; add an equal amount of 0.1% mercuric chloride, and shake and sterilize for 2.5 minutes Then suck it out (discard it in the special waste liquid tank); add the same amount of deionized water, shake and wash, suck out and put the seeds in a new EP tube, and repeat the water washing four times.

[0096] 2) Cultivation of Arabidopsis thaliana: put the...

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 liriodendron chinensis cellulose synthase gene LcCESA1 and as well as expression protein and application thereof. The nucleotide sequence of LcCESA1 is as shown in SEQ ID NO.1; the amino acid sequence of the expression protein is as shown in SEQ ID NO.2. By taking a model plant arabidopsis as a research object, a Chinese liriodendron chinensis cellulose synthase gene LcCESA1 obtained by cloning is mediated and transformed into arabidopsis, and a T3 generation homozygosous gene plant is finally obtained by means of screening and cultivation; by observing microstructures of the wild arabidopsis and transgenetic arabidopsis, and the condition that cell walls of histocytes of vascular bundles of stem sections and the blade histocytes of the transgenetic arabidopsis are different from those of the wild arabidopsis is found, so that the LcCESA1 gene affects the cell walls of histocytes of vascular bundles of stem sections and the blade histocytes of the transgeneticarabidopsis, can promote accumulation of cellulose and has wide application.

Description

technical field [0001] The invention belongs to the technical field of plant genes. Specifically related to a Liriodendron cellulose synthase gene LcCESA1 and its expression proteins and applications. Background technique [0002] Cellulose in plants is synthesized from polyprotein complexes, which are composed of hexamers in the plasma membrane, resembling a rosette structure. The single unit of each hexameric component is called cellulose synthase-A (cellulose synthase-A, CESA ), where the letter A represents the catalytic subunit. The basic structural unit of cellulose molecule is D-glucose pyranose, which is a high molecular polymer connected by β-1,4 glycosidic bonds, and forms a helical structure through intramolecular hydrogen bonds. Its glucose residues are about 2000-25000. Hemicellulose and pectin precursor oligomers are produced in the Golgi apparatus in cells, and then secreted to the plasma membrane by exocytosis to synthesize cellulose on the plasma membr...

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): C12N15/54C12N9/10A01H5/00A01H5/12A01H6/20
CPCC12N9/1059C12N15/8261
Inventor 陈金慧金磊磊施季森钟山成铁龙杨立明
Owner NANJING FORESTRY UNIV
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