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 Experiment Videos

Predicting evolution by in vitro evolution requires determining evolutionary pathways.

Barry G Hall1

  • 1Biology Department, University of Rochester, Rochester, New York 14627-0211, USA. drbh@mail.rochester.edu

Antimicrobial Agents and Chemotherapy
|August 17, 2002
PubMed
Summary
This summary is machine-generated.

Related Concept Videos

You might also read

Related Articles

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

Sort by
Same author

Building Phylogenetic Trees From Genome Sequences With kSNP4.

Molecular biology and evolution·2023
Same author

Estimating microbial population data from optical density.

PloS one·2022
Same author

Effects of sequence diversity and recombination on the accuracy of phylogenetic trees estimated by kSNP.

Cladistics : the international journal of the Willi Hennig Society·2021
Same author

Stress proteins as predictors of COVID-19 outcomes.

Cell stress & chaperones·2021
Same author

Statistical Package for Growth Rates Made Easy.

Molecular biology and evolution·2017
Same author

Population Dynamics of Staphylococcus aureus in Cystic Fibrosis Patients To Determine Transmission Events by Use of Whole-Genome Sequencing.

Journal of clinical microbiology·2017
Same journal

<i>In vitro</i> antibacterial activity of gepotidacin in combination with other antimicrobial agents against <i>Neisseria gonorrhoeae</i> isolates.

Antimicrobial agents and chemotherapy·2026
Same journal

Development of domain-specific probes of <i>Plasmodium falciparum</i> heat shock protein 70-1.

Antimicrobial agents and chemotherapy·2026
Same journal

Addressing therapeutic options for KPC-3-producing ST307-<i>Klebsiella pneumoniae</i>: insights from <i>in vitro</i> evolution and mutant prevention strategies.

Antimicrobial agents and chemotherapy·2026
Same journal

Indole-based hybrids target both asexual parasites and gametocytes of <i>Plasmodium falciparum</i> and synergize with lumefantrine.

Antimicrobial agents and chemotherapy·2026
Same journal

Lineage-specific loss of the type VI secretion system in <i>Acinetobacter baumannii</i> ST19 is associated with reduced accessory genome content.

Antimicrobial agents and chemotherapy·2026
Same journal

Genomic diversity and topical antimicrobial resistance in <i>Staphylococcus aureus</i> from clinical and carriage populations in the New York-New Jersey region.

Antimicrobial agents and chemotherapy·2026
See all related articles

DNA shuffling created a highly active TEM-1 beta-lactamase, but natural evolution likely follows a different path. A pathway exists for a less mutated enzyme, suggesting single mutations drive natural evolution of antibiotic resistance.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Evolutionary Biology

Background:

  • DNA shuffling can rapidly evolve enzymes with enhanced functions.
  • Natural evolution differs from in vitro evolution, with mutations typically arising sequentially.

Purpose of the Study:

  • To investigate the evolutionary pathway of TEM-1 beta-lactamase mutations.
  • To compare laboratory evolution via DNA shuffling with natural evolutionary processes.

Main Methods:

  • Site-directed mutagenesis was used to introduce individual amino acid substitutions from a DNA-shuffled mutant.
  • Repeated rounds of mutagenesis and selection were performed to build evolutionary pathways.
  • Enzyme activity was assessed to identify beneficial mutations.

Related Experiment Videos

Main Results:

  • No direct evolutionary pathway was found between wild-type TEM-1 and the six-mutation mutant generated by DNA shuffling.
  • A pathway was identified for a TEM-1 mutant with four substitutions, showing a fourfold increase in cefotaxime hydrolysis.
  • This four-substitution mutant's pathway suggests a single mutation could lead to a similar enzyme in nature.

Conclusions:

  • The evolutionary pathway for TEM-1 beta-lactamase activity differs between DNA shuffling and natural evolution.
  • Natural evolution of antibiotic resistance may be driven by single advantageous mutations, as exemplified by the TEM-52 allele.