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Revision as of 15:35, 28 October 2008

Bacillobricks: Introduction of BioBricks into Bacillus subtilis

Bacillus subtilis is potentially superior to E. coli as a host for some projects, for several reasons:

• It is much more effective at secreting proteins into the medium, as E. coli lacks the Main Terminal Branch of the General Secretory Pathway.
• As a Gram positive bacterium, it lacks the toxic lipopolysaccharide (endotoxin) of Gram negative bacteria such as 'E. coli
• The cells are considerably larger, making it easier to visualise intracellular components.
• B. subtilis forms endospores, a highly stable, heat and dessication resistant resting state which can be stored dry for years or decades, and will then germinate in less than 30 minutes when added to a suitable growth medium.
• B. subtilis is not pathogenic and has even been used as a probiotic organism in human foods.

However, standard BioBrick vectors do not allow introduction of BioBricks into B. subtilis. We felt that B. subtilis was potentially a suitable host for our 'Edinburgh Process' of conversion of cellulose to starch, due mainly to its ability to secrete enzymes such as cellulases. We therefore investigated two processes to allow introduction of BioBricks to B. subtilis, either on a plasmid or by integration into the genome. These experiments were carried out mainly by C. French (instructor) and Nimisha Joshi (advisor).

Introduction of BioBricks into B. subtilis on a plasmid

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