Integrating Membrane Transporter Proteins into Droplet Interface Bilayers

Heather E. Findlay, Nicola J. Harris, Paula J. Booth*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

1 Citation (Scopus)

Abstract

Droplet interface bilayers (DIBs) are an emerging tool within synthetic biology that aims to recreate biological processes in artificial cells. A critical component for the utility of these bilayers is controlled flow between compartments and, notably, uphill transport against a substrate concentration gradient. A versatile method to achieve the desired flow is to exploit the specificity of membrane proteins that regulate the movement of ions and transport of specific metabolic compounds. Methods have been in existence for some time to synthesize proteins within a droplet as well as incorporate membrane proteins into DIBS; however, there have been few reports combining synthesis and DIB incorporation for membrane transporters that demonstrate specific, uphill transport. This chapter presents two methods for the incorporation of a membrane transporter into a simple two-droplet DIB system, with the downhill and uphill transport reaction readily monitored by fluorescence microscopy.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc
Pages31-41
Number of pages11
DOIs
Publication statusPublished - 2021

Publication series

NameMethods in Molecular Biology
Volume2315
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Cell-free expression
  • Droplet interface bilayer
  • Fluorescence microscopy
  • Membrane protein
  • Transporter

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