JoVE Logo
Faculty Resource Center

Sign In

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

DOI :

10.3791/2505-v

9:01 min

March 16th, 2011

March 16th, 2011

29,690 Views

1Department of Microbiology & Environmental Toxicology, University of California Santa Cruz - UCSC

Here we demonstrate a simple protocol to create a random mutant library for a given target sequence. We show how this method, which is performed in vivo in Escherichia coli, can be coupled with functional selections to evolve new enzymatic activities.

Tags

Mutagenesis

-- Views

Related Videos

article

Transformation of Plasmid DNA into E. coli Using the Heat Shock Method

article

Homemade Site Directed Mutagenesis of Whole Plasmids

article

Crystallizing Membrane Proteins for Structure Determination using Lipidic Mesophases

article

Mutagenesis and Functional Analysis of Ion Channels Heterologously Expressed in Mammalian Cells

article

Examining the Conformational Dynamics of Membrane Proteins in situ with Site-directed Fluorescence Labeling

article

Use of a Robot for High-throughput Crystallization of Membrane Proteins in Lipidic Mesophases

article

High Throughput Quantitative Expression Screening and Purification Applied to Recombinant Disulfide-rich Venom Proteins Produced in E. coli

article

Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli

article

Functional Reconstitution and Channel Activity Measurements of Purified Wildtype and Mutant CFTR Protein

article

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

JoVE Logo

Privacy

Terms of Use

Policies

Research

Education

ABOUT JoVE

Copyright © 2024 MyJoVE Corporation. All rights reserved