Difference between revisions of "Part:BBa K2520004"

Revision as of 16:50, 26 October 2017

TRE-Mono display:MOG1-hGH

This device is a complete system for displaying proteins on cells' membrane. The Igk leader (BBa_K2520024) is a short signal peptide that prompts the translocation of a protein to the cellular membrane. PDGFR (BBa_K2520026) is a trans-membrane domain that anchors all the components located between the igk leader and the PDGFR itself to the membrane. HA (BBa_K2520030) is a tag that binds to specific antibodies and provide an indirect method for verifying the display of proteins on the membrane. The protein that we chose to express on the membrane is an epitope that is known target of multiple sclerosis (BBa_K2520038) and is component related to our specific project.

The TRE promoter (BBa_K2520025) is an inducible expression promoter in mammalian cells and hGH (BBa_K404108) serves as a terminator. The addition of TRE promoter allows inducible expression in specific conditions. We used the tet-off system. In the Tet-Off system, a tetracycline-controlled transactivator protein (tTA), which is composed of the Tet repressor DNA binding protein (TetR) from Escherichia coli fused to the strong transactivating domain of VP16 from Herpes simplex virus, regulates expression of a target gene that is under transcriptional control of a tetracycline-responsive promoter element (TRE).

The TRE is made up of Tet operator (tetO) sequence concatemers fused to a minimal promoter (commonly the minimal promoter sequence derived from the human cytomegalovirus (hCMV) immediate-early promoter). In the absence of Dox, tTA binds to the TRE promoter and activates transcription of the target gene. In the presence of Dox, tTA cannot bind to the TRE, and expression from the target gene remains inactive.

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
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 378
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 1045

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2520004 short</partinfo>
-This device is a complete system for displaying proteins on cells' membrane. The Igk leader (<partinfo>K2520024</partinfo>) is a short signal peptide that prompts the translocation of a protein to the cellular membrane. PDGFR (<partinfo>K2520026</partinfo>) is a trans-membrane domain that anchors all the components located between the igk leader and the PDGFR itself to the membrane. HA (<partinfo>K2520030</partinfo>) is a tag that binds to specific antibodies and provide an indirect method for verifying the display of proteins on the membrane. The protein that we chose to express on the membrane is an epitope that is known target of multiple sclerosis (<partinfo>K2520027</partinfo>) and is component related to our specific project.
+This device is a complete system for displaying proteins on cells' membrane. The Igk leader (<partinfo>K2520024</partinfo>) is a short signal peptide that prompts the translocation of a protein to the cellular membrane. PDGFR (<partinfo>K2520026</partinfo>) is a trans-membrane domain that anchors all the components located between the igk leader and the PDGFR itself to the membrane. HA (<partinfo>K2520030</partinfo>) is a tag that binds to specific antibodies and provide an indirect method for verifying the display of proteins on the membrane. The protein that we chose to express on the membrane is an epitope that is known target of multiple sclerosis (<partinfo>K2520038</partinfo>) and is component related to our specific project.
 The TRE promoter (<partinfo>K2520025</partinfo>) is an inducible expression promoter in mammalian cells and hGH (<partinfo>K404108</partinfo>) serves as a terminator. The addition of TRE promoter allows inducible expression in specific conditions. We used the tet-off system. In the Tet-Off system, a tetracycline-controlled transactivator protein (tTA), which is composed of the Tet repressor DNA binding protein (TetR) from Escherichia coli fused to the strong transactivating domain of VP16 from Herpes simplex virus, regulates expression of a target gene that is under transcriptional control of a tetracycline-responsive promoter element (TRE).