Recombinant vector over-expressing malic enzyme gene, and its application

A malic enzyme gene, recombinant vector technology, applied in the direction of recombinant DNA technology, microorganism-based methods, introduction of foreign genetic material using vectors, etc., can solve the problems of decreased ME activity, reduced ME activity, etc., to achieve favorable expression, high start-up Efficiency, size reduction effect

Inactive Publication Date: 2014-11-05
JINAN UNIVERSITY
View PDF2 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0015] On the contrary, studies have shown that the ME activity of Mucor circinelloide added with inhibitors is significantly reduced, because inhibitors can inhibit lipid metabolism and thus reduce ME activity (Kendrick, et al.1992)

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
  • Recombinant vector over-expressing malic enzyme gene, and its application
  • Recombinant vector over-expressing malic enzyme gene, and its application
  • Recombinant vector over-expressing malic enzyme gene, and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] 1. Construction of expression vector pHY18

[0057] 1. Analysis, cloning and connection of expression vector-related elements

[0058] The recombinant vector for overexpressing the malic enzyme gene of the present invention is obtained by transforming the commercialized vector pMD19-T (Takara, Dalian, China), and mainly has two expression cassettes, the resistance screening gene CAT expression cassette and the expression cassette of the target gene box.

[0059] The original expression control sequence was obtained by using NEB high-fidelity Pfu polymerase PCR and overlapping PCR (for specific PCR system and procedures, refer to Bryksin and Matsumura, 2010). The primers for the expression control sequence elements are: NR promoter (Pt13, Pt14), NR terminator (Pt79, Pt80). The CAT expression cassette was cloned from plasmid pHY11 with primers Pt83 and Pt84 (see Niu et al., 2012 for details). The PNR-PP3-SD fragment was amplified with primers Pt13 and Pt82. The Pt83, ...

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 recombinant vector over-expressing a malic enzyme gene, and its application. The recombinant vector comprises a screening labeled gene expression cassette and a target gene expression cassette; the screening labeled gene expression cassette comprises a resistance screening gene, the upstream of the resistance screening gene is fused with an Escherichia coli P1 promoter, and the downstream of the resistance screening gene is fused with an Escherichia coli ccdB terminator; and the target gene expression cassette comprises a Phaeodactylum tricornutum fcpC gene promoter, the malic enzyme gene, a Phaeodactylum tricornutum fcpA gene terminator and an Omega leader sequence, and the Omega leader sequence is in the downstream of the Phaeodactylum tricornutum fcpC gene promoter. The recombinant vector is small, is suitable for converting, is easy to operate, is a homologous sequence of the Phaeodactylum tricornutum, and benefits for expression. The over-expression of the malic enzyme gene can substantially improve the content of lipids in micro-algal cells, and lays the foundation for the researches of biological energy.

Description

Technical field: [0001] The invention belongs to the field of microalgae genetic engineering, and in particular relates to a recombinant vector for overexpressing malic enzyme gene and its application. Background technique: [0002] The non-renewability of traditional energy determines the trend of its depletion and causes increasingly serious environmental problems. The increasingly serious energy crisis and environmental pollution have aroused people's awareness of energy worries and the concept of zero pollutant discharge, strengthened the recycling and scientific utilization of waste oil and industrial waste, and promoted the development and utilization of clean new energy. Seeking a safe, reliable and clean renewable energy has become the focus of common attention in the world today. Bioenergy refers to the energy produced by using biorenewable raw materials and solar energy, including biomass energy, bioliquid fuel and energy produced by using biomass such as fuel alc...

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): C12N15/79C12N1/13C12R1/89
Inventor 李宏业薛姣杨维东刘洁生
Owner JINAN 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