AI Article Synopsis
- Developed a single-protein production system in E. coli using MazF, a toxin that selectively degrades cellular mRNAs, significantly reducing overall protein synthesis.
- When co-expressing MazF with a target gene devoid of ACA motifs, researchers achieved high levels of target protein expression (up to 90%) with minimal background noise.
- This technology also shows potential for producing various proteins, including those from yeast and human sources, facilitating easier structural and functional studies of challenging proteins.
Article Abstract
We designed a single-protein production (SPP) system in living E. coli cells that exploits the unique properties of MazF, a bacterial toxin that is an ssRNA- and ACA-specific endoribonuclease. In effect, MazF functions as an "mRNA interferase," because it efficiently and selectively degrades all cellular mRNAs in vivo, resulting in a precipitous drop in total protein synthesis. Concomitant expression of MazF and a target gene engineered to encode an ACA-less mRNA results in sustained and high-level (up to 90%) target expression in the virtual absence of background cellular protein synthesis. Remarkably, target synthesis continues for at least 4 days, indicating that cells retain transcriptional and translational competence despite their growth arrest. SPP technology works well for E. coli (soluble and membrane), yeast, and human proteins. This expression system enables unparalleled signal to noise ratios that should dramatically simplify structural and functional studies of previously intractable but biologically important proteins.
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| http://dx.doi.org/10.1016/j.molcel.2005.03.011 | DOI Listing |
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