Difference between revisions of "Part:BBa K2520023"

Revision as of 12:32, 26 October 2017

EF1a promoter

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 643
Illegal XhoI site found at 1042
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 777
Illegal AgeI site found at 151
1000
COMPATIBLE WITH RFC[1000]

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 <!-- Add more about the biology of this part here
 ===Usage and Biology===
+Human elongation factor-1 alpha (EF-1 alpha) is a constitutive promoter of human origin. It can be used in-vitro and in-vivo to induce ectopic expression of recombinant genes.  This promoter is very useful in cells where other promoters, such as CMV, are underactive or silenced (such as stem cells).
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-===Functional Parameters===
+===previous usage in iGEM===
+The EF-1a promoter was used by both teams that won “Best Therapeutics Project” last year (2016). This promoter allows for strong and constitutive expression in a wide variety of cell lines, and is especially useful when working with stem cells. This promoter has hitherto not been added to the iGEM parts registry as it included two forbidden restriction sites in its sequence. Since promoters are not translated, the base pair sequence cannot simply be changed through silent mutations. Since this promoter is clearly very important in iGEM, and specifically for therapeutics, we decided to mutate the original (WT) EF-1a at two points (base pair 319 and 824 were changed from C to T), thus eliminating the forbidden restriction sites. We characterized and tested our new promoter for functionality and compared it with the WT EF-1a (Figure 1). In closing we have added a new and invaluable promoter to the iGEM part registry that we believe will serve future teams seeking to work with mammalian cells.
+<!-- Uncomment this to enable Functional Parameter display
+===Results===
+In order to test our mutant promoter we created, we tested the expression of the reporter gene GFP under three different promoters- CMV, EF1a native and the mutant EF1a we created. As shown in the following graph, the expression levels of the reporter gene under EF1a promoter was much higher comparing to the CMV promoter, and these levels are almost equal between the native and the mutant promoter.
 <partinfo>BBa_K2520023 parameters</partinfo>
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