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

Application of glms gene in biosafety screening marker for lactic acid bacteria

A biosafety and screening marker technology, which is applied in the application field of glms gene in lactic acid bacteria biosafety screening markers, can solve the problems of large biological safety and hidden dangers, and achieve the effect of easy cloning, low cost and realization of homologous recombination

Inactive Publication Date: 2020-01-03
SHANGHAI ACAD OF AGRI SCI
View PDF4 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, some plasmid vectors and corresponding selection markers and other molecular elements used in lactic acid bacteria contain antibiotic resistance genes such as chloramphenicol and erythromycin, which bring great biological safety hazards

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
  • Application of glms gene in biosafety screening marker for lactic acid bacteria
  • Application of glms gene in biosafety screening marker for lactic acid bacteria
  • Application of glms gene in biosafety screening marker for lactic acid bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0044] The present invention will be further described below in conjunction with specific examples.

[0045] The source of the expression vector pUCERY: pUC19 is a laboratory-preserved plasmid, which was cloned and transformed to obtain pUCERY.

[0046] Example Construction of the glms gene-deficient strain of Lactococcus lactis L.lactis NZ9700

[0047] 1. Extraction and detection of genomic DNA of Lactococcus lactis

[0048] According to the literature (Ausubel F.M., Brent R.and Kingston R.E, et al.Short ProtocolsIn Molecular Biology.3rd ed.John Wiley&Sons, Inc., 1995), the Lactococcus lactis genomic DNA was extracted, and the specific process was as follows:

[0049] Cultivate 5ml of Lactococcus lactis bacteria solution to saturation, take 1.5ml of the bacteria solution and centrifuge for 2min; add 567μl of TE buffer to the precipitate, resuspend it by pipetting repeatedly, add 30μl of 10% SDS and 3μl of 20mg / ml Proteinase K, mix well, and incubate at 37°C for 1h; add 100μ...

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 provides an application of a glms gene in a biosafety screening marker for lactic acid bacteria. The glms gene is used as a biosafety screening marker for lactic acid bacteria, has no antibiotic pollution and no drug resistance diffusion, and is beneficial to the environment, convenient to screen and low in cost. The glms gene of the lactic acid bacteria is knocked out through homologous recombination to obtain a defective strain, the strain cannot survive without glucosamine and can normally grow with glucosamine, and growth condition of the strain cells is close to that of unknockout original strain. The glms gene, as the biosafety screening marker for lactic acid bacteria, has extremely high biological safety, is different from conventional selective marker genes derived from metabolic pathways, has advantages of convenient operation process, low cost, convenient screening, no limitation by culture medium components and the like, and provides a new important means forgenetic engineering of food microorganisms.

Description

technical field [0001] The invention belongs to the technical field of microbial genetic engineering, and in particular relates to the application of glms gene in biological safety screening markers of lactic acid bacteria. Background technique [0002] Glutamine, namely 6-phosphate fructose aminotransferase (L-Glutamine D-Fructose 6-phosphateAmidotransferase, GFAT, EC2.6.1.16), also known as 6-phosphate glucosamine synthetase (Glms), is an intracellular enzyme , Catalytic substrate glutamyl (glutamine) and 6-phosphate fructose (fructose-6-p) to form 6-phosphate glucosamine (glucosamine-6-p) and glutamic acid (glumate). [0003] GFAT catalyzes the first step reaction in the hexosamine synthesis metabolic pathway, and is the rate-limiting enzyme in this pathway. The catalyzed reaction is usually irreversible. The reaction catalyzed by GFAT consists of two combined enzymatic reactions. The first step reaction It is the hydrolysis of glutamine to produce a glutamic acid and a ...

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): C12N1/21C12R1/01
CPCC12N1/20C12N9/1096C12Y206/01016
Inventor 武国干唐雪明孙宇邓廷山曾海娟
Owner SHANGHAI ACAD OF AGRI SCI
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