IGEM:IMPERIAL/2008/New/Chassis 2

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Home The Project B.subtilis Chassis Wet Lab Dry Lab Notebook


Chassis 2 - Other pros, cons and information...

http://openwetware.org/wiki/IGEM:IMPERIAL/2008/Prototype/Project

Authors: Chris, James

Editors: Wiki team

James - Here is a table of potential advantages and disadvatages of using B.subtilis for our project and some more general chassis ad/disad. We may wish to think about chassis ad/dis seperatly, for example, in the registry the we will want to put the chassis info up!

Benefits
Challenges
Single Membrane - B.subtilis is a gram-positive bacterium and contains only a single membrane. This makes B.subtilis an ideal organism for secretion of proteins (in our case, peptides for the biomaterials) in high amounts. Starting from scratch - Although previous iGEM teams have looked at B.subtilis as a potential chassis, there are few working B.subtilis parts. As a result we will have to design a vaerity of promoter, ribosome binding sites and protein coding regions.
Availability of Potential Parts - B.subtilis is a highly studied organism, its genome has been fully sequenced and many of these genes have been functionaly annotated. As a result, B.subtilis provides many useful potential parts that we could take advantage of. For example within our project, potential parts for control of motility, light sensing and secretion pathways.
Non-pathogenicity - B.subtilis is a non-pathogenic organism that is commonly found in soil. As a result, B.subtilis has a biological harzardous level of 1 and offers a useful non-pathogenic chassis for synthetic biologists. Peptidase activity - B.subtilis possesses a stringent protein folding checking system that may reject our biomaterials. Proteins expressed in B.subtilis may not fold correctly causing a build up inside the cell. B.subtilis responds to this problem by expressing exopeptidases, which digest the protein. There is a risk that our biomaterial may be cleaved up before it can be secreted.
Naturally Competent - B. subtilis has been noted for its ease and efficieny of transformation. Plasmids can be naturally taken up by B. subtilis cells. Vector Degradation - B.subtilis does not use all the same vectors as E.coli. One reason for this is that B.subtilis often recognises vectors grown in E.coli and digests them. This will (judging by the approach of previous iGEM teams) include the biobrick itself.
High Motility - B.subtilis is often referred to as a highly motile organsim. Quoted velocity range from 60 - 100 μm per second. In comparison to E.coli which has an average velocity of .... B.subtilis is much more motile.
Sporulation: Transport - Under stress conditions, B.subtilis will form spores. These spores are highly resistant versions of single cells, able to withstand extreme temperatures and pH. These spores are capable of growing into new cells once favourable growing conditions are restored. Due to the resistance of spores, there is a great potential for manipulating them for transporting B.subtilis devices and constructs.

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