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Related Concept Videos

Restriction Enzymes01:11

Restriction Enzymes

Restriction enzymes are bacterial enzymes used to cut DNA in a sequence-specific manner. To cleave DNA, they bind to specific palindromic sequences called restriction sites. Such palindromic DNA sequences or inverted repeats are commonly found in regions of functional significance, such as the origin of replication, gene operator sites, and regions containing transcription termination signals.
The host bacteria protect their own genomic DNA from these enzymes by methylating these sites. Some...
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
Evolution of New Traits in Microbes01:24

Evolution of New Traits in Microbes

Microorganisms evolve rapidly due to their large population sizes and short generation times, often exhibiting measurable changes within days under laboratory conditions. Natural selection acts on standing genetic variation, enabling the retention and amplification of beneficial traits that confer fitness advantages in changing environments.Adaptive Pigment Regulation in RhodobacterIn Rhodobacter, a genus of purple non-sulfur bacteria, light-harvesting pigments such as bacteriochlorophyll and...

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Related Experiment Video

Updated: May 24, 2026

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
09:16

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity

Published on: March 25, 2020

Restriction endonucleases: natural and directed evolution.

Richa Gupta1, Neena Capalash, Prince Sharma

  • 1Department of Biotechnology, Panjab University, Chandigarh, India 160014.

Applied Microbiology and Biotechnology
|March 9, 2012
PubMed
Summary
This summary is machine-generated.

Engineered restriction enzymes (REs) offer novel DNA cleavage specificities crucial for molecular biology. Directed evolution and protein engineering enable the creation of customized REs with enhanced properties for diverse applications.

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A New Screening Method for the Directed Evolution of Thermostable Bacteriolytic Enzymes

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Enzymology

Background:

  • Type II restriction endonucleases (REs) are highly sequence-specific nucleases.
  • The PD-(D/E)XK nuclease superfamily, including REs, is diverse, sharing a conserved core but little sequence similarity.
  • REs are classified by function, not genetic relatedness, due to limited sequence homology.

Purpose of the Study:

  • To explore the evolution and engineering of restriction enzymes (REs) for novel DNA specificities.
  • To highlight advancements in characterizing and designing REs with tailored properties.
  • To address the increasing demand for new RE specificities in molecular biology.

Main Methods:

  • Development of new alignment matrices and classification systems for REs.
  • Analysis of protein evolution through mutations, recombination, insertions, and deletions.
  • Application of rational mutagenesis and directed evolution for protein redesign.
  • Engineering strategies including domain combination and modification of amino acid sequences.

Main Results:

  • Over 300 distinct RE specificities are cataloged in REBASE.
  • Engineered REs demonstrate altered cleavage specificity, substrate preference, and stability.
  • A growing number of patents are awarded for engineered REs with improved functions.

Conclusions:

  • Restriction enzyme engineering is vital for advancing molecular biology tools.
  • Directed evolution and protein redesign offer powerful approaches to create novel REs.
  • The continuous evolution of REs meets the increasing demand for specific DNA targeting capabilities.