Document Detail


Inducible expression of transmembrane proteins on bacterial magnetic particles in Magnetospirillum magneticum AMB-1.
MedLine Citation:
PMID:  20038711     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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 tetracycline-inducible 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 tetracycline-inducible expression system described here will be a useful tool for the expression and display of transmembrane proteins in the membranes of BacMPs.
Authors:
Tomoko Yoshino; Akiko Shimojo; Yoshiaki Maeda; Tadashi Matsunaga
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2009-12-28
Journal Detail:
Title:  Applied and environmental microbiology     Volume:  76     ISSN:  1098-5336     ISO Abbreviation:  Appl. Environ. Microbiol.     Publication Date:  2010 Feb 
Date Detail:
Created Date:  2010-02-08     Completed Date:  2010-04-02     Revised Date:  2010-09-28    
Medline Journal Info:
Nlm Unique ID:  7605801     Medline TA:  Appl Environ Microbiol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1152-7     Citation Subset:  IM    
Affiliation:
Department of Biotechnology, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588, Japan. y-tomoko@cc.tuat.ac.jp
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MeSH Terms
Descriptor/Qualifier:
Antigens, CD / biosynthesis,  genetics
Bacterial Proteins / biosynthesis*,  genetics*
Base Sequence
DNA Primers / genetics
Genes, Reporter
Genetic Vectors
Green Fluorescent Proteins / biosynthesis,  genetics
Lipid Bilayers
Magnetics
Magnetosomes / metabolism
Magnetospirillum / genetics*,  metabolism*
Membrane Proteins / biosynthesis*,  genetics*
Plasmids / genetics
Protein Engineering
Recombinant Proteins / biosynthesis,  genetics
Tetracycline / pharmacology
Viral Envelope Proteins / genetics,  metabolism
Chemical
Reg. No./Substance:
0/Antigens, CD; 0/Bacterial Proteins; 0/CD81 antigen; 0/DNA Primers; 0/Lipid Bilayers; 0/Membrane Proteins; 0/Recombinant Proteins; 0/Viral Envelope Proteins; 147336-22-9/Green Fluorescent Proteins; 157184-61-7/glycoprotein E2, Hepatitis C virus; 60-54-8/Tetracycline
Comments/Corrections

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