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

T4 DNA ligase variants with increased salt tolerance

A ligase, mutant technology, applied in the field of T4 DNA ligase variants, can solve the problems of sample loss, increase time and complexity, and achieve high activity effect

Active Publication Date: 2022-07-08
WUHAN AIBO TAIKE BIOTECH CO LTD
View PDF7 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Such added purification steps add time and complexity to the experiment, and the purification procedure often results in sample loss - which can be significant

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
  • T4 DNA ligase variants with increased salt tolerance
  • T4 DNA ligase variants with increased salt tolerance
  • T4 DNA ligase variants with increased salt tolerance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0012] The term "biologically active fragment" refers to any fragment, derivative, homologue, or analog of a T4 DNA ligase mutant that has an in vivo or in vitro activity characteristic of a biomolecule; including, for example, ligase activity, or via a ligation repair band Mismatches present in nicked DNA. In some embodiments, the biologically active fragment, derivative, homologue or analog of the mutant T4 DNA ligase has any degree of biological activity of the mutant T4 DNA ligase in any in vivo or in vitro assay of interest .

[0013] In some embodiments, the biologically active fragment can optionally include any number of contiguous amino acid residues of a mutant T4 DNA ligase. The present invention also includes polynucleotides encoding any such biologically active fragments.

[0014] Biologically active fragments can result from post-transcriptional processing or translation of alternatively spliced ​​RNAs, or alternatively can be produced by engineering, batch syn...

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

A variety of T4DNA ligase mutants that exhibit enhanced ligation activity at high salt concentrations as compared to wild-type ligases are engineered, characterized and selected via gel electrophoresis of ligation products from standard ligation assays. A ligase catalyzes the formation of a phosphodiester bond between the 5'end and the 3 'end of complementary cohesive or blunt ends of double helix DNA, which process is crucial for many molecular biological processes, including cloning and sequencing.

Description

Background technique [0001] DNA ligases are indispensable repair proteins in living organisms that catalyze the repair of single-strand breaks in double-stranded DNA. T4 DNA ligase uses the energy of the biomolecule ATP to repair nicked double-stranded DNA, and also ligates double-stranded DNA with complementary single-stranded overhangs (sticky ends) as well as blunt-ended fragments, making it a useful tool for in vitro combinatorial smaller Useful tool for DNA fragments to generate double-stranded DNA constructs. [0002] DNA ligase forms phosphodiester bonds between duplex nucleic acid fragments at the intersection of juxtaposed 5' phosphate and 3' hydroxyl termini. By designing complementary overhangs between each double-stranded fragment, ligation can be both position-specific and directional. This allows the specific integration of DNA or RNA fragments into larger vectors to meet the needs of molecular biology research. Among commercially available ligases, T4 DNA lig...

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): C12N9/00C12N15/52C12N15/10
CPCC12N9/93C12Y605/01001C12N15/102C12Q2525/307C12Q2521/501C12Q2527/125C12Q1/6869
Inventor 朱振宇孙大鹏
Owner WUHAN AIBO TAIKE BIOTECH CO LTD
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