Difference between revisions of "Part:BBa K1431301"

Revision as of 13:29, 17 October 2014

TRE-3G promoter+SV40 PolyA, an ideal controller of mammalian gene expression with Tet-On 3G protein

TRE-3G promoter is a kind of eukaryocyte cell promoters that will be switched on after combine with the compound of Tet-On 3G(BBa_K1431101) protein and doxycycline(Dox) mix.
The inducible promoter PTRE3G provides for very low basal expression and high maximal expression after induction (Loew et. al., 2010). It consists of 7 repeats of a 19 bp tet operator sequence located upstream of a minimal CMV promoter. In the presence of Dox, Tet-On 3G binds specifically to P_TRE3G and activates transcription of the downstream GOI. P_TRE3G lacks binding sites for endogenous mammalian transcription factors, so it is virtually silent in the absence of induction. That is a very useful way to control whether a protein express or not.

There are two BbsI sites between promoter and PolyA which can make the coding sequence seamless gather with them. So we can also test how many nucleotides interval between promoter and coding sequence will make the highest expression efficiency.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

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 <partinfo>BBa_K1431301 short</partinfo>
-TRE-3G promoter is a kind of eukaryocyte cell promoters that will be switched on after combine with the compound of Tet-On 3G([https://parts.igem.org/Part:BBa_K1431101 BBa_K1431101]) protein and doxycycline mix.
+TRE-3G promoter is a kind of eukaryocyte cell promoters that will be switched on after combine with the compound of Tet-On 3G([https://parts.igem.org/Part:BBa_K1431101 BBa_K1431101]) protein and doxycycline(Dox) mix. <br>
-That is a very useful way to control whether protein express or not. Of course PolyA is needful in eukaryocyte cell. There are two BbsI sites between promoter and PolyA which can make the coding sequence seamless gather with them. So we can also test how many nucleotides interval between promoter and coding sequence will make the highest expression efficiency.
+The inducible promoter PTRE3G provides for very low basal expression and high maximal expression after induction (Loew et. al., 2010). It consists of 7 repeats of a 19 bp tet operator sequence located upstream of a minimal CMV promoter. In the presence of Dox, Tet-On 3G binds specifically to P<sub>TRE3G</sub> and activates transcription of the downstream GOI.  P<sub>TRE3G</sub> lacks binding sites for endogenous mammalian transcription factors, so it is virtually silent in the absence of induction.
+That is a very useful way to control whether a protein express or not.
+ There are two BbsI sites between promoter and PolyA which can make the coding sequence seamless gather with them. So we can also test how many nucleotides interval between promoter and coding sequence will make the highest expression efficiency.
 <!-- Add more about the biology of this part here