Plastid production of protein antibiotics against pneumonia via a new strategy for high-level expression of antimicrobial proteins

Oey, Melanie, Lohse, Marc, Scharff, Lars B., Kreikemeyer, Bernd and Bock, Ralph (2009) Plastid production of protein antibiotics against pneumonia via a new strategy for high-level expression of antimicrobial proteins. Proceedings of the National Academy of Sciences of the United States of America, 106 16: 6579-6584. doi:10.1073/pnas.0813146106


Author Oey, Melanie
Lohse, Marc
Scharff, Lars B.
Kreikemeyer, Bernd
Bock, Ralph
Title Plastid production of protein antibiotics against pneumonia via a new strategy for high-level expression of antimicrobial proteins
Journal name Proceedings of the National Academy of Sciences of the United States of America   Check publisher's open access policy
ISSN 1091-6490
Publication date 2009-04-21
Sub-type Article (original research)
DOI 10.1073/pnas.0813146106
Open Access Status
Volume 106
Issue 16
Start page 6579
End page 6584
Total pages 16
Place of publication Washington, DC, United States
Publisher National Academy of Sciences
Language eng
Abstract Plastid transformation has become an attractive tool in biotechnology. Because of the prokaryotic nature of the plastid's gene expression machinery, expression elements (promoters and untranslated regions) that trigger high-level foreign protein accumulation in plastids usually also confer high expression in bacterial cloning hosts. This can cause problems, for example, when production of antimicrobial compounds is attempted. Their bactericidal activity can make the cloning of the corresponding genes in plastid transformation vectors impossible. Here, we report a general solution to this problem. We have designed a strategy (referred to as toxin shuttle) that allows the expression in plastids of proteins that are toxic to Escherichia coli. The strategy is based on blocking transcription in E. coli by bacterial transcription terminators upstream of the gene of interest, which subsequently are excised in planta by site-specific recombination. We demonstrate the applicability of the strategy by the high-level expression in plastids (to up to 30% of the plant's total soluble protein) of 2 phage-derived protein antibiotics that are toxic to E. coli. We also show that the plastid-produced antibiotics efficiently kill pathogenic strains of Streptococcus pneumoniae, the causative agent of pneumonia, thus providing a promising strategy for the production of next-generation antibiotics in plants.
Keyword Chloroplast
Molecular farming
Plastid transformation
Phage lysin
Site-specific recombination
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: Institute for Molecular Bioscience - Publications
 
Versions
Version Filter Type
Citation counts: TR Web of Science Citation Count  Cited 58 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 64 times in Scopus Article | Citations
Google Scholar Search Google Scholar
Created: Fri, 31 Oct 2014, 12:07:03 EST by Melanie Oey on behalf of Institute for Molecular Bioscience