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Please use this identifier to cite or link to this item: http://hdl.handle.net/10204/1037

Title: Development of a flagellin surface display expression system in a moderate thermophile, Bacillus halodurans Alk36
Authors: Crampton, M
Berger, E
Reid, S
Louw, M
Keywords: Flagellin
Cell surface display
Bacillus halodurans
Fusion protein
Integration
Issue Date: Jun-2007
Publisher: Springer Verlag
Citation: Crampton, M, Berger, E, Reid, S and Louw, M. 2007. Development of a flagellin surface display expression system in a moderate thermophile, Bacillus halodurans Alk36. Applied Microbiology and Biotechnology, Vol 75(3), pp 599-607
Abstract: This study relates to the development of an alkaliphilic, thermotolerant, Gram-positive isolate, Bacillus halodurans Alk36, for the over-production and surface display of chimeric gene products. This bacterium continuously over-produces flagellin. To harness this ability, key genetic tools, such as gene targeted inactivation, were developed for this strain. The hag gene, which codes for flagellin, was inactivated on the chromosome giving rise to the B. halodurans BhFC01 mutant. Polylinkers were inserted as in-frame, chimeric, flagellin sandwich fusions to identify the permissive insertion sites corresponding to the variable regions of the flagellin protein. Flagellin expression and motility were evaluated for these constructs. Two sites were identified for possible peptide insertion in the flagellin gene, one of which produced functional flagella and was able to restore the motility phenotype to a non-motile mutant. Peptides encoding a poly-histidine peptide and the HIV-1 subtype C gp120 epitope were, respectively, incorporated into this site as in-frame fusions. The peptides were found to be successfully displayed on the cell surface and functional through metal binding and immunological studies, respectively
Description: Copyright: 2007 Springer Verlag
URI: http://hdl.handle.net/10204/1037
ISSN: 0175-7598
Appears in Collections:Microbial expression systems
General science, engineering & technology

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