"
Team:Pitt/Protocol Design/Results
From 2014.igem.org
Results
As of 10/16/2014, 12 of 16 trials have currently been completed with no positive results (Table 1). The largest hurdle to completing trails was the fact that 4 of the strains that were grown at 24°C did not ever reach the optical density that was required for an attempt at transformation. In addition, the other 4 strains grown at 24°C that did reach an O/D of around 0.8 took much longer than the 2 weeks that it took for the strains grown in the 37°C room. Since, we did not have 16 trails, we could not perform a DOX analysis to determine which variables were the most important for transformation. This led us to conclude that if another DOX analysis is conducted with these parameters, it would be efficacious to use a low value for culture temperature that is higher than 24°C in order to at least have viable P. Acnes to perform a transformation protocol with.
Table 1
| Date: | Std/Run | Results: |
| 8/8/14 | 11/16 | Lawn, phage test showed it was not p. acnes |
| 8/8/14 | 15/3 | 17 colonies, colony PCR showed that they didn’t contain the plasmid |
| 8/8/14 | 3/11 | 73 colonies, colony PCR showed that they didn’t contain the plasmid |
| 9/2/14 | 9/10 | No growth after electroporation |
| 9/2/14 | 5/9 | No growth after electroporation |
| 9/2/14 | 1/2 | No growth after electroporation |
| 9/23/14 | 4/5 | 3 colonies, not p. acnes by color and growth pattern |
| 9/23/14 | 6/13 | No growth after electroporation |
| 9/23/14 | 7/6 | 1 colony, not p. acnes by color and growth pattern |
| 9/23/14 | 13/15 | No growth after electroporation |
| 9/23/14 | 10/8 | No growth after electroporation |
| 9/23/14 | 16/14 | No growth after electroporation |
The goal of our part of the experiment was to develop a transformation protocol for a bacteria that was on the skin. This would allow us to incorporate a plasmid into the bacteria that could produce a protein to reduce acne. P. Acnes was the obvious choice since it is abundant on the skin and aggressive strains of the bacteria are associated with acne formation. As we discovered, however, it is rather difficult to transform. This led us to consider other bacteria on the skin that are easier to transform.
We came up with a number of candidates for transformation and identified transformation protocols. The information below indicates which bacteria are the best for transformation:
Top candidates ideal skin location and transformation protocol available: S. aureus, S. epidermidis, M. lysodeikticus.
Top candidates ideal skin location but no transformation protocols found by search: P. acnes, P. acidifaciens, P. australiense, P. avidum, P. granulosum, P. humerusii, P. lymphophilum, P. propionicum, C. amicolatum, C. striatum, C. jeikeium, C. urealyticum, and C. xerosis.
Conclusions
Page currently under Construction.
Next Page
Previous Page
