Team:Groningen/Navigation/Attributions

From 2013.igem.org

Attributions

We have thought of this project by ourselves, so it was not an assignment from one of our supervisors. The supervisors and advisors main role was providing consultancy. All the design, labwork and modeling was done by the team itself and started in June. We also took care of the sponsoring, other organizational issues and made our own presentations and poster.

The silk that we have developed was based on the article of Brooks et al. [1], but the codon optimization was executed by the team itself using software written by one of the team members. We improved the backbone made by the iGEM team from Munich in 2012 and used this backbone for our constructs. Also, we validated that biobrick BBa_E0840 works in Bacillus subtilis (Figure 1).

The coating mechanism was developed by the team using article [2][3][4]. The knockout strains were shipped from the university of Illinois and Buenos Aires .

Figure 1: The intensity in of ~40 cells in the GFP-channel, were analyzed from two pictures. The average intensity (AU) from the cells are plotted above with the standard deviation. A GFP from our group is shown and in a biobricked GFP (BBa_E0840) is shown in a plot. wt=wildtype, T0= before induction, T1,2,3=#h of induction with IPTG.

References

1: Brooks, Amanda E., Shane M. Stricker, Sangeeta B. Joshi, Timothy J. Kamerzell, C. Russell Middaugh, and Randolph V. Lewis. "Properties of Synthetic Spider Silk Fibers Based on Argiope Aurantia MaSp2." Biomacromolecules 9.6 (2008): 1506-510.
2: Garrity, Liam F., and George W. Ordal. "Chemotaxis in Bacillus subtilis: How bacteria monitor environmental signals." Pharmacology & therapeutics 68.1 (1995): 87-104.
3: Bredeston, L. M., et al. "Thermal regulation of membrane lipid fluidity by a two‚Äźcomponent system in Bacillus subtilis." Biochemistry and Molecular Biology Education 39.5 (2011): 362-366.
4: Rao, Christopher V., George D. Glekas, and George W. Ordal. "The three adaptation systems of Bacillus subtilis chemotaxis." Trends in microbiology 16.10 (2008): 480-487.