Difference between revisions of "Team:MIT/Experiments/Promoters/Design-Descisions"
m |
|||
| (2 intermediate revisions by 2 users not shown) | |||
| Line 13: | Line 13: | ||
-We additionally liked the fact that the synthetic pEREs drastically shortened the length of promoter from the 2000+ base pairs of pTFF1. | -We additionally liked the fact that the synthetic pEREs drastically shortened the length of promoter from the 2000+ base pairs of pTFF1. | ||
-We also decided to try to synthesize an inducible promoter rather than doing the fusion with VP16-GAL4 (like the progesterone side) since we wanted to maintain and utilize as many natural components of the cell itself. | -We also decided to try to synthesize an inducible promoter rather than doing the fusion with VP16-GAL4 (like the progesterone side) since we wanted to maintain and utilize as many natural components of the cell itself. | ||
| + | </p> | ||
| + | <h2 style = "text-decoration:underline; font-family: Trebuchet MS;">Progesterone Promoters</h2> | ||
| + | <p style = "font-family:Verdana;"> Lorem ipsum dolor sit amet, consectetur adipiscing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. Duis aute irure dolor in reprehenderit in voluptate velit esse cillum dolore eu fugiat nulla pariatur. Excepteur sint occaecat cupidatat non proident, sunt in culpa qui officia deserunt mollit anim id est laborum. | ||
| + | </p> | ||
| + | <h2 style = "text-decoration:underline; font-family: Trebuchet MS;">Estrogen-Progesterone Hybrid Promoter</h2> | ||
| + | <p style = "font-family:Verdana;"> Lorem ipsum dolor sit amet, consectetur adipiscing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. Duis aute irure dolor in reprehenderit in voluptate velit esse cillum dolore eu fugiat nulla pariatur. Excepteur sint occaecat cupidatat non proident, sunt in culpa qui officia deserunt mollit anim id est laborum. | ||
</p> | </p> | ||
</body> | </body> | ||
</html> | </html> | ||
Latest revision as of 20:16, 13 October 2016
Design Decisions for Hormone Inducible Promoters
Estrogen Promoters
-ERE's are spaced within the locations of the TetO sites of the TRE inducible promoter. Additionally, since the tetO site is 19 base pairs and ERE is only 13 base pairs, we added 5 additional random and different base pairs upstream, directly in front of the ERE sequence, to maintain the geometry of the TRE promoter since the way the DNA folds is important and we didn't want the DNA to misfold by having an incorrect length. -ERE sequence has three blank base pairs in the middle of its sequence, which we chose to fill randomly and differently for each occurence of an ERE site since there is no indication that they have any effect on the functionality of the binding to ER-alpha and also because IDT would not synthesize it with so many repeated sequences. -Chose repetitions of 3,5, and 6 because the paper had most success with 3 synthetic EREs, then five was a middle number, and 6 is the number of TetO sites on TRE-tight that we wanted to mimic since that was a successful synthetically built inducible promoter. -Choose pTFF1 because that is a natural promoter that responds to ER-alpha. -We additionally liked the fact that the synthetic pEREs drastically shortened the length of promoter from the 2000+ base pairs of pTFF1. -We also decided to try to synthesize an inducible promoter rather than doing the fusion with VP16-GAL4 (like the progesterone side) since we wanted to maintain and utilize as many natural components of the cell itself.
Progesterone Promoters
Lorem ipsum dolor sit amet, consectetur adipiscing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. Duis aute irure dolor in reprehenderit in voluptate velit esse cillum dolore eu fugiat nulla pariatur. Excepteur sint occaecat cupidatat non proident, sunt in culpa qui officia deserunt mollit anim id est laborum.
Estrogen-Progesterone Hybrid Promoter
Lorem ipsum dolor sit amet, consectetur adipiscing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. Duis aute irure dolor in reprehenderit in voluptate velit esse cillum dolore eu fugiat nulla pariatur. Excepteur sint occaecat cupidatat non proident, sunt in culpa qui officia deserunt mollit anim id est laborum.
