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Updated: Jun 15, 2026

Protocols for Implementing an Escherichia coli Based TX-TL Cell-Free Expression System for Synthetic Biology

Protocols for Implementing an Escherichia coli Based TX-TL Cell-Free Expression System for Synthetic Biology

Published on: September 16, 2013

Development of an efficient expression system for Flavobacterium strains.

Shicheng Chen¹, Michael G Kaufman, Michael Bagdasarian

¹Department of Microbiology and Molecular Genetics, Michigan State University, East Lansing, MI 48824, USA. shicheng@msu.edu

|March 9, 2010

Summary

This summary is machine-generated.

Researchers identified strong Flavobacterium johnsoniae promoters to express fluorescent proteins and insecticidal genes in Flavobacterium. These engineered bacteria showed mosquito larval toxicity, highlighting potential for biocontrol applications.

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Last Updated: Jun 15, 2026

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Cell-free Protein Expression Using the Rapidly Growing Bacterium Vibrio natriegens

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Published on: March 14, 2019

Area of Science:

Microbiology
Molecular Biology
Biotechnology

Background:

Flavobacterium strains are important in various environments but lack well-characterized expression systems.
Efficient genetic tools are needed for functional genomics and biotechnological applications in Flavobacteria.

Purpose of the Study:

To isolate and characterize strong promoters from Flavobacterium johnsoniae.
To develop expression systems for fluorescent proteins and insecticidal genes in Flavobacterium species.
To assess the potential of engineered Flavobacteria for mosquito biocontrol.

Main Methods:

Promoter-trap vector system using a green fluorescent protein (GFP) reporter.
Isolation and sequence analysis of promoter DNA fragments from Flavobacterium johnsoniae.
Cloning and expression of fluorescent protein markers and insecticidal genes (Bacillus sphaericus binary toxins) in Flavobacterium strains.
Testing toxicity of engineered Flavobacteria against mosquito larvae.

Main Results:

Identified conserved Bacteroidetes promoter motifs (TTG-N(19)-TAnnTTTG) in F. johnsoniae.
Isolated potent promoters, including PompA, that drive strong gene expression in Flavobacterium hibernum but not in Escherichia coli.
Demonstrated successful co-expression of fluorescent proteins and insecticidal proteins in Flavobacterium.
Flavobacteria engineered to express Bacillus sphaericus p42 and p51 toxins showed significant toxicity against larvae of Culex quinquefasciatus, Anopheles gambiae, and Ochlerotatus triseriatus.
Constitutive expression of Bacillus thuringiensis cry11A gene was found to be problematic in Flavobacterium.

Conclusions:

Novel strong promoters from F. johnsoniae have been characterized for use in Flavobacterium expression systems.
Engineered Flavobacteria expressing insecticidal toxins show promise as agents for mosquito larval control.
Further optimization is needed for expressing certain heterologous genes, such as cry11A, in Flavobacterium.