Flexible Programming of Cell-free Protein Synthesis Using Magnetic Bead-immobilized Plasmids

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2012 Apr 4

PMID 22470570

Citations 7

Authors

Ka-Young Lee

Kyung-Ho Lee

Ji-Woong Park

Dong-Myung Kim

Affiliations

Soon will be listed here.

Abstract

The use of magnetic bead-immobilized DNA as movable template for cell-free protein synthesis has been investigated. Magnetic microbeads containing chemically conjugated plasmids were used to direct cell-free protein synthesis, so that protein generation could be readily programmed, reset and reprogrammed. Protein synthesis by using this approach could be ON/OFF-controlled through repeated addition and removal of the microbead-conjugated DNA and employed in sequential expression of different genes in a same reaction mixture. Since the incubation periods of individual template plasmids are freely controllable, relative expression levels of multiple proteins can be tuned to desired levels. We expect that the presented results will find wide application to the flexible design and execution of synthetic pathways in cell-free chassis.

Citing Articles

Solid-Phase Cell-Free Protein Synthesis and Its Applications in Biotechnology.

Sanchez-Costa M, Lopez-Gallego F Adv Biochem Eng Biotechnol. 2023; 185:21-46.

PMID: 37306703 DOI: 10.1007/10_2023_226.

Efficient multi-gene expression in cell-free droplet microreactors.

Restrepo Sierra A, Arold S, Grunberg R PLoS One. 2022; 17(3):e0260420.

PMID: 35312702 PMC: 8936439. DOI: 10.1371/journal.pone.0260420.

Loss of a single methylation in 23S rRNA delays 50S assembly at multiple late stages and impairs translation initiation and elongation.

Wang W, Li W, Ge X, Yan K, Mandava C, Sanyal S Proc Natl Acad Sci U S A. 2020; 117(27):15609-15619.

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MEERCAT: Multiplexed Efficient Cell Free Expression of Recombinant QconCATs For Large Scale Absolute Proteome Quantification.

Takemori N, Takemori A, Tanaka Y, Endo Y, Hurst J, Gomez-Baena G Mol Cell Proteomics. 2017; 16(12):2169-2183.

PMID: 29055021 PMC: 5724179. DOI: 10.1074/mcp.RA117.000284.

A molecular nanodevice for targeted degradation of mRNA during protein synthesis.

Lee K, Min S, Kim H, Lee S, Kim D Sci Rep. 2016; 6:20733.

PMID: 26857021 PMC: 4746582. DOI: 10.1038/srep20733.

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