Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

6.2K
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...
6.2K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Research on rural co-governance model under "micro-regeneration of community spaces" - a case study of Shanghai Pujiang country park.

Scientific reports·2026
Same author

Stellar feedback drives the baryon deficiency in low-mass galaxies.

Science advances·2026
Same author

Altered thalamic subregional connectivity with default mode and frontoparietal networks in adolescents with depression.

Journal of affective disorders·2026
Same author

Therapeutic targeting of lysosome-triggered inflammatory channels in nasal and upper-airway allergic conditions.

Frontiers in immunology·2026
Same author

Integrative Taxonomy Reveals a Candidate Lineage Within the <i>Rhinolophus macrotis</i> Group.

Biology·2026
Same author

Pressurized Gas-Driven Elemental Redistribution Enables Ultrastable PtNi Catalysts for Heavy-Duty Vehicles.

Journal of the American Chemical Society·2026

Related Experiment Video

Updated: Oct 3, 2025

Directed Evolution Method in Saccharomyces cerevisiae: Mutant Library Creation and Screening
10:50

Directed Evolution Method in Saccharomyces cerevisiae: Mutant Library Creation and Screening

Published on: April 1, 2016

11.1K

Hot spots-making directed evolution easier.

Haoran Yu1, Shuang Ma1, Yiwen Li2

  • 1Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China; Hangzhou Global Scientific and Technological Innovation Centre, Zhejiang University, Hangzhou 311200, China.

Biotechnology Advances
|February 13, 2022
PubMed
Summary
This summary is machine-generated.

Directed evolution enhances proteins by intelligently selecting mutation sites, known as hot spots. This strategy, using sequence or structure data, creates efficient "smart libraries" for targeted protein engineering.

Keywords:
Directed evolutionEnzyme engineeringHot spotsTarget mutagenesis

More Related Videos

A New Screening Method for the Directed Evolution of Thermostable Bacteriolytic Enzymes
13:30

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

Published on: November 7, 2012

18.2K
Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli
09:01

Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli

Published on: March 16, 2011

30.7K

Related Experiment Videos

Last Updated: Oct 3, 2025

Directed Evolution Method in Saccharomyces cerevisiae: Mutant Library Creation and Screening
10:50

Directed Evolution Method in Saccharomyces cerevisiae: Mutant Library Creation and Screening

Published on: April 1, 2016

11.1K
A New Screening Method for the Directed Evolution of Thermostable Bacteriolytic Enzymes
13:30

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

Published on: November 7, 2012

18.2K
Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli
09:01

Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli

Published on: March 16, 2011

30.7K

Area of Science:

  • Protein engineering
  • Biotechnology
  • Computational biology

Background:

  • Directed evolution is a key method for protein engineering.
  • Traditional library construction methods (error-prone PCR, DNA shuffling) are often inefficient.
  • High-throughput screening is frequently lacking, making library searching laborious.

Purpose of the Study:

  • To review mutation target site (hot spot) prediction methods for directed evolution.
  • To categorize hot spots based on sequence and 3D structure information.
  • To highlight computational tools and successful applications in enzyme engineering.

Main Methods:

  • Overview of seven types of hot spots: sequence-based (CbD sites, coevolving residues) and structure-based (active-site, access tunnel, flexible, distal, interface residues).
  • Discussion of computational tools for identifying these hot spots.
  • Compilation of successful case studies in enzyme engineering.

Main Results:

  • Hot spot prediction is critical for the efficiency and success of directed evolution.
  • Smart libraries, generated using predicted hot spots, increase the ratio of beneficial mutations.
  • Categorization provides a framework for understanding and selecting optimal mutation sites.

Conclusions:

  • Accurate prediction of mutation hot spots significantly improves the efficiency of directed evolution.
  • Utilizing sequence and structure-based hot spots enables rational and effective protein engineering.
  • Advances in computational tools facilitate the identification of hot spots for successful enzyme engineering.