Difference between revisions of "Team:WPI Worcester/Results"
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Curcumin’s antibacterial ability against different strains of bacteria was assessed. To see if it had a blanket response among different bacterial species was an objective of this study. | Curcumin’s antibacterial ability against different strains of bacteria was assessed. To see if it had a blanket response among different bacterial species was an objective of this study. | ||
Observing the data, particularly the mismatch in drops and increases in biofilm measurement a few reasons for this occurrence arises. One concerns the very development of biofilm initially, wherein, when placed under taxing, harsh conditions nonideal for development, some bacteria form biofilm (Kostakioti, 2013). Curcumin, posited in the literature to kill bacteria, may have proven so harsh for bacteria with membrane deconstructing mechanism that cells went into biofilm development to endure, explaining the increase in biofilm detected in S. aureus for example. Another reason may concern the measurement technique used. While great care was taken to be consistent among all assayed plates in all trials, there was no standardization in the initial wash step of the plates to apply the crystal violet stain, which, on those occasions the water was applied for longer than needed, removing the biofilm, thus skewing data. | Observing the data, particularly the mismatch in drops and increases in biofilm measurement a few reasons for this occurrence arises. One concerns the very development of biofilm initially, wherein, when placed under taxing, harsh conditions nonideal for development, some bacteria form biofilm (Kostakioti, 2013). Curcumin, posited in the literature to kill bacteria, may have proven so harsh for bacteria with membrane deconstructing mechanism that cells went into biofilm development to endure, explaining the increase in biofilm detected in S. aureus for example. Another reason may concern the measurement technique used. While great care was taken to be consistent among all assayed plates in all trials, there was no standardization in the initial wash step of the plates to apply the crystal violet stain, which, on those occasions the water was applied for longer than needed, removing the biofilm, thus skewing data. | ||
Revision as of 17:48, 12 October 2018
Results
Biofilm Quantification
Antifreeze Proteins against Biofilm Formation
HIStag Purification
Protein Concentration Assay
SDS Page Gel
Biofilm Assay Result
Curcumin Assays
As can be seen, the EMG strain had the most pronounced biofilm development. While DH5α had an apparent drop in biofilm development, it like S. epidermidis and S. aureus showed an increase in biofilm growth. NCTC similarly shows an initial drop but an uptick in measured biofilm after 20 µM curcumin solution was applied. However, overlap between error bars in these four aforementioned strains among ratios means overall measurements may be suspect.
Discussion and Future Work
Curcumin’s antibacterial ability against different strains of bacteria was assessed. To see if it had a blanket response among different bacterial species was an objective of this study. Observing the data, particularly the mismatch in drops and increases in biofilm measurement a few reasons for this occurrence arises. One concerns the very development of biofilm initially, wherein, when placed under taxing, harsh conditions nonideal for development, some bacteria form biofilm (Kostakioti, 2013). Curcumin, posited in the literature to kill bacteria, may have proven so harsh for bacteria with membrane deconstructing mechanism that cells went into biofilm development to endure, explaining the increase in biofilm detected in S. aureus for example. Another reason may concern the measurement technique used. While great care was taken to be consistent among all assayed plates in all trials, there was no standardization in the initial wash step of the plates to apply the crystal violet stain, which, on those occasions the water was applied for longer than needed, removing the biofilm, thus skewing data. Future extensions of this project could involve running this experiment once more with the aforementioned issues addressed and remedied. Moreover, discerning a scalable method conducive for industrial production of curcumin from synthetic bacteria (like what is executed currently with insulin) may prove insightful and provide for a case study for a new industry of standardized herbal biologics. If desired, the genes comprising the mechanism of curcumin could be spliced into plant genomes to have the plants produce curcumin themselves.Lettuce Colony Counts
Gene Gun Transformation Results
Transformation with Antifreeze Proteins
Project Achievements
We achieved a lot over the course of our project, both positive and negative results. Below are our lists of successes and failures.
Successes of ICEberg
- Successful Purification of Antifreeze proteins in BL21 strains of E. coli
- Reduction of Biofilm formation with any application of
- Transformation of EMG cells to express Antifreeze protein
- Production of a functional and inexpensive Gene Gun
- Transformed lettuce leaves to express GFP using the team Gene Gun
Failures of ICEberg
- Inability to make competent NCTC cells with multiple protocols
- IPTG inducible operon did not turn on