Difference between revisions of "Part:BBa K1075018"

Latest revision as of 02:45, 5 October 2013

J23105-RBS32-SspB[Core]-LOV-ipaA-TT

This part contains the sspB[Core] domain fused to the LOV-ipaA construct with a constitutive promoter, ribosomal binding site and a double terminator.

Usage and Biology

ipaA forms a tight dimer with Vincolin. LOV is a light-sensitive protein domain which was fused to ipaA. That way the dimerization of ipaA and Vincolin was rendered light-induced.

EcsspB itself regulates the degradation of ssrA tagged proteins through the ClpXP protease in procaryotes. In engineered systems it is used to induce degradation of specifically ssrA tagged proteins. The XB domain of sspB is part of the split system of E. Coli sspB (EcsspB) which was used in fusion with FKBP and FRB to induce the activity of sspB and therefore degradation of proteins with Rapamycins. [1] EcsspB can be functionally divided in three parts: -the N terminal Core domain (113 AA) -the C terminal XB peptide (25 AA) -and a 'flexible linker' in between the first parts ( We used the same domain structure as is used within the Rapamycin inducable split system. While the Core domain is responsible for dimerization of sspB and binding of ssrA the XB domain binds the protease ClpXP. The flexible linker is called flexible because it was found that an increase or reduction in size or amino acid composition does not influence the function of sspB as much as it would in the other domains.

The part can be used to regulate the dimerization of the two parts of the split sspB. The binding of ipaA to Vincolin is regulated via the light-sensitive LOV domain. Therefore the two sspB parts only come together when the LOV domain was activated with blue light.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 7
Illegal NheI site found at 30
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 1: / Line 1: @@
 __NOTOC__
 <partinfo>BBa_K1075018 short</partinfo>
-a
+This part contains the sspB[Core] domain fused to the LOV-ipaA construct with a constitutive promoter, ribosomal binding site and a double terminator.
+===Usage and Biology===
+ipaA forms a tight dimer with Vincolin.
+LOV is a light-sensitive protein domain which was fused to ipaA. That way the dimerization of ipaA and Vincolin was rendered light-induced.
+EcsspB itself regulates the degradation of ssrA tagged proteins through the ClpXP protease in procaryotes. In engineered systems it is used to induce degradation of specifically ssrA tagged proteins.
+The XB domain of sspB is part of the split system of E. Coli sspB (EcsspB) which was used in fusion with FKBP and FRB to induce the activity of sspB and therefore degradation of proteins with Rapamycins. [1] EcsspB can be functionally divided in three parts:
+-the N terminal Core domain (113 AA)
+-the C terminal XB peptide (25 AA)
+-and a 'flexible linker' in between the first parts ( We used the same domain structure as is used within the Rapamycin inducable split system. While the Core domain is responsible for dimerization of sspB and binding of ssrA the XB domain binds the protease ClpXP. The flexible linker is called flexible because it was found that an increase or reduction in size or amino acid composition does not influence the function of sspB as much as it would in the other domains.
+The part can be used to regulate the dimerization of the two parts of the split sspB. The binding of ipaA to Vincolin is regulated via the light-sensitive LOV domain. Therefore the two sspB parts only come together when the LOV domain was activated with blue light.
 <!-- Add more about the biology of this part here