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

Phaeodactylum tricomutum diacylglycerol acyltransferase and encoding gene and application thereof

A technology of diacylglycerol and Phaeodactylum sp. is applied in the field of genetic engineering, which can solve the problems of unsatisfactory and complicated oil content, and achieve the effect of promoting development and improving oil content.

Inactive Publication Date: 2013-04-10
SICHUAN UNIV
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, it is a very complicated work to screen out algae species with high algae oil content and suitable for local environment cultivation from these microalgae
[0005] Phaeodactylum tricornutum is a marine microalgae that can be artificially cultivated on a large scale at present, and its oil content reaches 20-30%. There is still a long way to go for the industrial application of algae energy
[0006] At present, several DAGAT-encoding cDNAs have been identified in these organisms mouse (Mus musculus), Arabidopsis thaliana, tobacco (Nicotiana tabacum), but there are few reports on the actual production and application of transgenic plants, especially the use of DAGAT to cultivate There is no relevant report on the genetic engineering breeding of algae with high oil content

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
  • Phaeodactylum tricomutum diacylglycerol acyltransferase and encoding gene and application thereof
  • Phaeodactylum tricomutum diacylglycerol acyltransferase and encoding gene and application thereof
  • Phaeodactylum tricomutum diacylglycerol acyltransferase and encoding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Molecular Cloning of Ptdagat Gene from Phaeodactylum tricornutum

[0024] The acquisition of the full-length cDNA of Phaeodactylum tricornutum diacylglycerol transferase (Ptdagat) gene mainly includes the following steps:

[0025] Using diacylglycerol transferase (AY054480) from Arabidopsis thaliana in the Phaeodactylum tricornutum genome database that has completed whole genome sequencing (the USDepartment of Energy Joint Genome Institute (http: / / www.jgi.doe.gov / ) In Blast analysis, a partial sequence encoding diacylglycerol transferase was found, and a pair of primers were designed according to the sequence:

[0026] P1 (upstream primer), 5'-CACGCTACGATA ACA TGGATGAA-3';

[0027] P2 (downstream primer), 5'-CAATCTTTGTAGAACACACGATCTC-3';

[0028] The total RNA of Phaeodactylum tricornutum in logarithmic growth phase was extracted (RNA extract kit, Watson, Shanghai, China), and the RNA was used as a template to synthesize cDNA with AMV reverse transcriptase (TaKaRa, Da...

Embodiment 2

[0030] Expression of Ptdagat Encoding Gene in Several Different Growth Conditions of Phaeodactylum tricornutum

[0031] figure 1 Among them, A means to cultivate Phaeodactylum tricornutum with normal silicon-containing f / 2 medium, and B means to cultivate Phaeodactylum tricornutum with f / 2 medium without silicon element nutrient salt. Depend on figure 1 From A and B, it can be seen that the expression level of Ptdagat gene in the former is not as high as that in the latter. By extracting and analyzing the oil content of the two groups, the result is that the oil content of the latter is correspondingly higher. This shows that the expression level of the gene of the present invention is positively correlated with the oil content of Phaeodactylum tricornutum. figure 1 C, D, E, and F in the table indicate that different concentrations of abscisic acid ABA (0mg / L, 1mg / L, 5mg / L, 15mg / L) were added to the medium of Phaeodactylum tricornutum. Concentrations of ABA caused differenc...

Embodiment 3

[0033] Antisense suppression of Ptdagat-encoding gene expression and RNA interference caused significant changes in lipid content in genetically engineered algae

[0034] Antisense RNA (antisense) suppression of Ptdagat gene in Phaeodactylum tricornutum: Using the 34th to 984th base of the Ptdagat coding gene as a material, a total of 951 nucleotide sequences were used to reverse clone this sequence into Phaeodactylum tricornutum Algae expression vectors such as the multiple cloning site of pPha-T1, thus constructing an antisense RNA expression vector that is reverse complementary to the Ptdagat coding gene. With a gene gun ( PDS-1000 / He Particle Delivery System) was used to transform Phaeodactylum tricornutum, positive clones were screened with Zeocin antibiotics, and then positive clones were further detected by PCR, and the correctness of the structure was analyzed by sequencing. Grease content. The experimental results showed that: the antisense RNA inhibited the expres...

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 Phaeodactylum tricomutum diacylglycerol acyltransferase, an encoding gene and application thereof, and belongs to the technical field of genetic engineering. An amino acid residue sequence of the Phaeodactylum tricomutum diacylglycerol acyltransferase is shown as SEQ ID No: 2. The gene for encoding the Phaeodactylum tricomutum diacylglycerol acyltransferase is one of the following nucleotide sequences: 1, SEQ ID No: 1 in a sequence table, 2, a nucleotide sequence for encoding a protein sequence shown as SEQ ID No: 2 in the sequence table, and 3, a deoxyribonucleic acid(DNA) sequence which has more than 90 percent of homology with a nucleotide sequence defined in the SEQ ID No: 1 in the sequence table and encodes the same functional protein. Through the experiment,the transgenosis can remarkably improve the content of grease in algae, so that a high-quality gene is provided for improving the content of the grease in the algae; and the Phaeodactylum tricomutum diacylglycerol acyltransferase has broad application prospect and important application value.

Description

technical field [0001] The invention relates to a plant gene technology, in particular to a Phaeodactylum tricornutum diacylglycerol transferase (Ptdagat) related to the synthesis of triacylglycerides and its coding gene and application, belonging to the technical field of genetic engineering. Especially the construction of genetically engineered algae strains involving high oil content energy microalgae. Background technique [0002] Energy is the lifeblood of national economic and social development. With the depletion of fossil energy resources, the price of crude oil has been rising all the way, and the combustion of fossil energy has led to the CO in the atmosphere 2 As a result of the net increase in production and the deterioration of the environment and climate, countries around the world have to consider accelerating the pace of research and development of petroleum alternative raw materials, among which biodiesel is regarded as a renewable alternative energy sourc...

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 Patents(China)
IPC IPC(8): C12N9/10C12N15/54C12N15/63C12N5/10C12N15/79C12N15/82C12R1/91
Inventor 卿人韦涂丽兰利琼曲芳兵李湘汪乐霓傅华龙
Owner SICHUAN 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