Difference between revisions of "Team:East Chapel Hill/Parts"
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<img src="https://static.igem.org/mediawiki/2018/f/f1/T--East_Chapel_Hill--PROMOTER.png"</img> | <img src="https://static.igem.org/mediawiki/2018/f/f1/T--East_Chapel_Hill--PROMOTER.png"</img> | ||
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| − | <p2 style= "font-size: 18-x;"> This part is an improvement of the 2017 East Chapel Hill iGem "CHOP" operon. At fluoride concentrations of 2uM and above, the delta-CRCB E coli strain (strain JW0619-1 from the Yale Coli Genetics Stock Center) will grow on chloramphenicol. This vector is designed with Xhol cutting sites so one may easily remove and replace different segments of the operon to screen for combinations of riboswitch and promoter with highest effectivity. | + | <p2 style= "font-size: 18-x;"> This part is an improvement of the 2017 East Chapel Hill iGem "CHOP" operon. At fluoride concentrations of 2uM and above, the <i>delta</i>-<i>CRCB E. coli</i> strain (strain JW0619-1 from the Yale Coli Genetics Stock Center) will grow on chloramphenicol. This vector is designed with Xhol cutting sites so one may easily remove and replace different segments of the operon to screen for combinations of riboswitch and promoter with highest effectivity. |
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To summarize, here is how this part functions in the presence of fluoride: | To summarize, here is how this part functions in the presence of fluoride: | ||
| − | <br> 1. <I> delta-CRCB E.coli</I> accumulates fluoride intracellularly due to the lack of the fluoride efflux channel. | + | <br> 1. <I> delta</I>-<I>CRCB E.coli</I> accumulates fluoride intracellularly due to the lack of the fluoride efflux channel. |
<br> 2. Fluoride binds to the fluoride riboswitch and activates the chloramphenicol acetyltransferase enzyme, which allows for bacterial growth on plates that contain the antibiotic chloramphenicol. | <br> 2. Fluoride binds to the fluoride riboswitch and activates the chloramphenicol acetyltransferase enzyme, which allows for bacterial growth on plates that contain the antibiotic chloramphenicol. | ||
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<p2 style= "font-size: 18-x;"> | <p2 style= "font-size: 18-x;"> | ||
| − | This is BBa_K2843002 operon with the "Fluoride Binding Mutant", which is a mutant of the fluoride RBS. This serves as a controlled experiment because if there is growth of <I> delta-CRCB E. coli </I> on the chloramphenicol plates than this indicates that growth of<i> delta</i>-<i>CRCB</i> is not entirely dependent only on the presence of fluoride.</p2> | + | This is BBa_K2843002 operon with the "Fluoride Binding Mutant", which is a mutant of the fluoride RBS. This serves as a controlled experiment because if there is growth of <I> delta</I>-<I>CRCB E.coli </I> on the chloramphenicol plates than this indicates that growth of<i> delta</i>-<i>CRCB</i> is not entirely dependent only on the presence of fluoride.</p2> |
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Revision as of 02:47, 18 October 2018
Parts
To summarize, here is how this part functions in the presence of fluoride:
1. delta-CRCB E.coli accumulates fluoride intracellularly due to the lack of the fluoride efflux channel.
2. Fluoride binds to the fluoride riboswitch and activates the chloramphenicol acetyltransferase enzyme, which allows for bacterial growth on plates that contain the antibiotic chloramphenicol.
