Inducible expression of transmembrane proteins on bacterial magnetic particles in magnetospirillum magneticm AMB-1

Tomoko Yoshino, Akiko Shimojo, Yoshiaki Maeda, Tadashi Matsunaga

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Bacterial magnetic particles (BacMPs) produced by the magnetotactic bacterium Magnetospirillum magneticum AMB-1 are used for a variety of biomedical applications. In particular, the lipid bilayer surrounding BacMPs has been reported to be amenable to the insertion of recombinant transmembrane proteins; however, the display of transmembrane proteins in BacMP membranes remains a technical challenge due to the cytotoxic effects of the proteins when they are overexpressed in bacterial cells. In this study, a tetracyclineinducible expression system was developed to display transmembrane proteins on BacMPs. The expression and localization of the target proteins were confirmed using luciferase and green fluorescent protein as reporter proteins. Gene expression was suppressed in the absence of anhydrotetracycline, and the level of protein expression could be controlled by modulating the concentration of the inducer molecule. This system was implemented to obtain the expression of the tetraspanin CD81. The truncated form of CD81 including the ligand binding site was successfully displayed at the surface of BacMPs by using Mms13 as an anchor protein and was shown to bind the hepatitis C virus envelope protein E2. These results suggest that the tetracyclineinducible expression system described here will be a useful tool for the expression and display of transmembrane proteins in the membranes of BacMPs.

Original languageEnglish
Pages (from-to)1152-1157
Number of pages6
JournalApplied and Environmental Microbiology
Volume76
Issue number4
DOIs
Publication statusPublished - 2010 Feb
Externally publishedYes

Fingerprint

Magnetospirillum
transmembrane proteins
Bacterial Proteins
protein
Proteins
proteins
lipid bilayers
Hepatitis C virus
Viral Envelope Proteins
luciferase
green fluorescent protein
recombinant proteins
Lipid Bilayers
cytotoxicity
binding sites
Green Fluorescent Proteins
Luciferases
Recombinant Proteins
Hepacivirus
protein synthesis

ASJC Scopus subject areas

  • Biotechnology
  • Food Science
  • Applied Microbiology and Biotechnology
  • Ecology

Cite this

Inducible expression of transmembrane proteins on bacterial magnetic particles in magnetospirillum magneticm AMB-1. / Yoshino, Tomoko; Shimojo, Akiko; Maeda, Yoshiaki; Matsunaga, Tadashi.

In: Applied and Environmental Microbiology, Vol. 76, No. 4, 02.2010, p. 1152-1157.

Research output: Contribution to journalArticle

Yoshino, Tomoko ; Shimojo, Akiko ; Maeda, Yoshiaki ; Matsunaga, Tadashi. / Inducible expression of transmembrane proteins on bacterial magnetic particles in magnetospirillum magneticm AMB-1. In: Applied and Environmental Microbiology. 2010 ; Vol. 76, No. 4. pp. 1152-1157.
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