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

Difunctional thiolase gene MvaE, expression vector, recombinant strain and application thereof

A thiolase and dual-function technology, applied in the direction of carrier, application, nucleic acid carrier, etc., can solve problems that have not been completely solved

Active Publication Date: 2021-03-26
扬州麦塔瑞思生物科技有限公司
View PDF1 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the key technology that restricts the microbial synthesis efficiency of fatty acids has not been completely resolved.

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
  • Difunctional thiolase gene MvaE, expression vector, recombinant strain and application thereof
  • Difunctional thiolase gene MvaE, expression vector, recombinant strain and application thereof
  • Difunctional thiolase gene MvaE, expression vector, recombinant strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: The artificial design and synthesis of the bifunctional thiolase gene MvaE based on Escherichia coli codon bias

[0029] According to the DNA sequence of the MvaE gene shown in SEQ ID NO: 1, the codon of the MvaE gene was optimized to facilitate its expression in Escherichia coli. The optimized nucleic acid sequence is shown in SEQ NO: 2 after adding the ribosome binding site sequence. The codon-optimized gene sequence was synthesized by Sangon Bioengineering (Shanghai) Co., Ltd., and the 5' and 3' ends of the synthesized gene sequence had BamH I and Hind III restriction sites, respectively.

Embodiment 2

[0030] Embodiment 2: The construction of the expression vector pSTmvaE that is used for Escherichia coli expression MvaE gene

[0031] A DNA sequence such as SEQ NO: 2 is constructed on an expression vector, such as pSTV28 vector. The specific implementation steps are as follows:

[0032] The DNA fragment synthesized in Example 1 and the carrier plasmid pSTV28 were double-digested with BamH I and Hind III respectively; the digested DNA fragment and the large vector fragment were recovered by agarose electrophoresis, and the digested gene fragments were sequentially placed at 16 by DNA ligase. ℃ Ligated to the corresponding digested product of pSTV28 to form pSTmvaE. Escherichia coli MG1655 was transformed with each ligation reaction product, and then spread on a petri dish containing 30 μg / mL Cmp (chloramphenicol), and incubated at 37° C. for 10-12 hours. The next day, a single colony was picked to extract the plasmid using the Shanghai Sangon plasmid mini-extraction kit. T...

Embodiment 3

[0033] Example 3 Construction of Escherichia coli recombinant bacteria overexpressing bifunctional thiolase gene MvaE

[0034] The pSTV28 and pSTmvaE plasmids in Example 2 were respectively electrotransformed into Escherichia coli MG1655 to obtain the Escherichia coli engineering strains MG: pSTV28 and pSMG: pSTmvaE respectively.

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 difunctional thiolase gene MvaE. The invention further discloses difunctional thiolase, an expression cassette, a recombinant vector, recombinant bacteria or cells and application thereof. The invention further discloses a method for producing fatty acid. The difunctional thiolase gene MvaE disclosed by the invention can utilize rich culture media containing glucose, glycerol or other carbon sources as raw materials under facultative anaerobic conditions, so that the fatty acid content of C<14:0>, C<16:0> and C<18:0> in host bacteria can be remarkably increased, and the increasing amplitude of the difunctional thiolase gene MvaE is different from 27% to 213% according to different host strains.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a bifunctional thiolase gene MvaE, an expression vector thereof, a recombinant bacterial strain and applications thereof. Background technique [0002] Fatty acids are important platform chemicals that can be widely used in the synthesis of biofuels, organic detergents, rubber compounds, plastic products, textile chemicals, and food additives. The traditional ways to obtain fatty acids include direct extraction and separation from animal and plant cells and chemical synthesis from petroleum products. These methods have disadvantages such as low efficiency, high cost, high land occupation rate and unfriendly environment. Compared with traditional methods, microbial synthesis of fatty acids has inherent advantages in terms of cost, land occupation rate and environmental protection, and is considered to be the most potential method that can replace traditional methods. Howe...

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): C12N15/54C12N15/53C12N9/10C12N9/04C12N15/70C12N1/21C12P7/64C12R1/19
CPCC12N9/1029C12N9/0006C12Y101/01034C12N15/70C12P7/6409C12N2800/22
Inventor 周佳田宝霞李相前
Owner 扬州麦塔瑞思生物科技有限公司
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