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

Firefly luciferase gene and application thereof

A luciferase gene and luciferase technology, applied in the application field of the gene in the production of firefly luciferase, can solve the problems of affecting enzyme activity, difficult separation, low enzyme activity, etc., and achieve easy purification and high enzyme purity , highly active effect

Active Publication Date: 2015-02-04
GENOMEDI TECH SHANGHAI
View PDF1 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Jeffrey constructed the recombinant vector pkw101 containing the firefly luciferase gene in 1985. Production using the system constructed by Jeffrey requires heat induction at 45 degrees for 30 minutes, which seriously affects the enzyme activity. Other production methods also have difficulties in separation. Low yield, low enzyme activity and other serious problems

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
  • Firefly luciferase gene and application thereof
  • Firefly luciferase gene and application thereof
  • Firefly luciferase gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0020] The present invention will be described in detail below with reference to the accompanying drawings and in combination with embodiments.

[0021] A firefly luciferase gene is a mutLuc gene obtained by mutating the 31st lysine of the wild-type firefly luciferase gene into alanine.

[0022] Further, the nucleotide sequence of the wild-type firefly luciferase gene is shown in SEQ.ID.NO.1, and the amino acid sequence of the alanine is shown in SEQ.ID.NO.2.

[0023] A firefly luciferase gene is used in the production of firefly luciferase.

[0024] see figure 1 As shown, in the Pet-mutLuc, since the firefly luciferase mutation is under the control of the high-efficiency promoter T7 gene and the lactose operon Lac, a certain concentration of isopropyl-b-D-thiogalactoside (IPTG ) exists, the firefly luciferase gene of the present invention can be efficiently induced to transcribe.

[0025] Utilize described mutLuc gene to load the Xho1 I of recombinant expression vector pet...

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 firefly luciferase gene and an application thereof. The firefly luciferase gene is a mutLuc gene which is obtained through mutating the 31st-bit lysine of a wild type firefly luciferase gene into alanine and is used for producing firefly luciferase. Compared with a convention firefly luciferase production method, the wild type firefly luciferase gene is mutated and a Luc codon is optimized, so that the mutated firefly luciferase has the characteristics of easiness in purification, high yield (each litre of zymocyte can obtain more than 5mg of apoenzyme), high enzymic purity (enzymic specific activity is not less than 1*10<11> remaining useful life (RUL) / mg protein), high activity, strong stability and the like; and the method can efficiently produce the firefly luciferase, and is suitable for industrial production and applied to individual diagnosis.

Description

technical field [0001] The invention relates to a new firefly luciferase gene, in particular to the application of the gene in producing firefly luciferase. Background technique [0002] Luciferase is a general term for enzymes that can produce bioluminescence in nature, the most representative of which is the luciferase in a firefly named Phot inus pyralis. In the corresponding chemical reaction, the generation of fluorescence comes from the oxidation of luciferin, and in some cases, adenosine triphosphate (ATP) is also included in the reaction system. In the absence of luciferase, luciferin reacts very slowly with oxygen, and the presence of calcium ions often speeds up the reaction further (similar to what happens with muscle contraction). The chemical reaction is as follows: [0003] Luciferin+ATP+O2→Oxyluciferin+PPi+AMP+CO2+light. This reaction is so energy-efficient that almost all of the energy that goes into the reaction is converted into light. In the presence o...

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): C12N15/53C12N9/02
Inventor 李润生
Owner GENOMEDI TECH SHANGHAI
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