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

Gene editing vector for corynebacterium glutamicum, preparation method, system and application thereof

A technology of gene editing and glutamic acid bar, applied in the field of gene editing, can solve the problems of time-consuming efficiency and low efficiency, and achieve the effect of short cycle and cost-saving efficiency

Inactive Publication Date: 2017-11-24
JIANGNAN UNIV
View PDF4 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Currently, there are only a few vectors for gene editing in Corynebacterium glutamicum, such as pKl8mobsacB / pKl9mobsacB, but the above-mentioned vectors are time-consuming and inefficient because they require two homologous recombination to achieve gene editing

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
  • Gene editing vector for corynebacterium glutamicum, preparation method, system and application thereof
  • Gene editing vector for corynebacterium glutamicum, preparation method, system and application thereof
  • Gene editing vector for corynebacterium glutamicum, preparation method, system and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Embodiment 1: A gene editing system of Corynebacterium glutamicum

[0050] 1. A gene editing vector of Corynebacterium glutamicum, the nucleotide sequence is as shown in SEQ ID No.4, and its plasmid map is as shown in figure 1 as shown,

[0051] Gene editing vectors include Cas9 expression elements, sgRNA expression elements, replicon elements and homologous repair sequence insertion sites;

[0052] The cas9 expression elements include cat chloramphenicol resistance gene, lactose operon lacIq, promoter Ptac, SD sequence, cas9 sequence and terminator rrnB, and SD sequence is connected before the initiation codon ATG of the cas9 sequence;

[0053] The sgRNA expression element includes promoter Ptrc, sgRNA scaffold sequence and terminator T1T2, the sgRNA scaffold sequence includes trancrRNA and crRNA insertion sites, and the crRNA insertion site is an AjuI restriction site;

[0054] The replicon elements include the C. glutamicum replicon repA and the Escherichia coli r...

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 a gene editing vector for corynebacterium glutamicum. The gene editing vector has higher gene editing efficiency. The gene editing vector comprises a Cas9 expression element, an sgRNA expression element, a replicon element and a homologous repairing sequence insertion site, wherein the Cas9 expression element comprises a resistance gene, an operon, a promoter, a SD sequence, a cas9 sequence and a terminator; the cas9 sequence is shown as SEQ ID No.5; the SD sequence is shown as SEQ ID No.6; the sgRNA expression element comprises a promoter, an sgRNA scaffold sequence and a terminator; the sgRNA scaffold sequence comprises trancrRNA and crRNA insertion sites; the sgRNA scaffold sequence is shown as SEQ ID No.7; AND the replicon element comprises a corynebacterium glutamicum replicon repA.

Description

technical field [0001] The invention relates to the field of gene editing, in particular to a gene editing vector, preparation method, system and application of Corynebacterium glutamicum. Background technique [0002] For industrial microorganisms, it is necessary to undergo metabolic analysis and optimization from wild-type strains to efficient production strains, and use gene editing technology to genetically modify and rationally transform the host. Gene editing technology refers to the use of exogenous DNA to mutate the target gene of the recipient cell genome through recombination, thereby generating the target mutant strain. Gene editing requires genome operations such as gene knockout, gene insertion, and gene replacement. [0003] CRISPR / Cas (Clustered regularly interspaced short palindromic repeats, CRISPR) / Cas9 system is an acquired immune system for bacteria and archaea to deal with the invasion of foreign DNA. Through RNA-mediated, Cas9 protein can specifically...

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): C12N15/77C12N9/22C12N15/10C12N15/66
CPCC12N9/22C12N15/102C12N15/66C12N15/77
Inventor 刘秀霞彭枫王新月孙杨杨艳坤白仲虎
Owner JIANGNAN 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