Abstract
Cell-free transcription–translation (TXTL) has recently emerged as a versatile technology to engineer biological systems. In this chapter, we show how an all E. coli TXTL system can be used to build synthetic cell prototypes. We describe methods to encapsulate TXTL reactions in cell-sized liposomes, with an emphasis on the composition of the external solution and lipid bilayer. Cell-free expression is quantitatively described in bulk reactions and liposomes for three proteins: the soluble reporter protein eGFP, the membrane proteins alpha-hemolysin (AH) from Staphylococcus aureus, and the mechanosensitive channel of large conductance (MscL) from E. coli.
| Original language | English (US) |
|---|---|
| Title of host publication | Methods in Enzymology |
| Publisher | Academic Press Inc. |
| Pages | 217-239 |
| Number of pages | 23 |
| Volume | 617 |
| DOIs | |
| State | Published - 2019 |
Publication series
| Name | Methods in enzymology |
|---|---|
| Publisher | Academic Press Inc. |
| ISSN (Print) | 0076-6879 |
Bibliographical note
Publisher Copyright:© 2019 Elsevier Inc.
Keywords
- Bioengineering
- Cell-free transcription–translation
- Gene circuits
- Liposomes
- Membrane proteins
- Synthetic biology
- Synthetic cell
- Protein Biosynthesis
- Escherichia coli/genetics
- Hemolysin Proteins/genetics
- Genetic Engineering/methods
- Bacterial Toxins/genetics
- Green Fluorescent Proteins/genetics
- Ion Channels/genetics
- Escherichia coli Proteins/genetics
- Transcription, Genetic
- Liposomes/metabolism
- Staphylococcus aureus/genetics
- Artificial Cells/metabolism
PubMed: MeSH publication types
- Research Support, Non-U.S. Gov't
- Research Support, U.S. Gov't, Non-P.H.S.
- Journal Article
