Difference between revisions of "Part:BBa K1075009"

Latest revision as of 02:19, 5 October 2013

E. coli sspB[XB]

This part contains the XB part of the split version of sspB.

Usage and Biology

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 peptide 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.

There is no start codon included in this part, because regulatory parts are fused N-terminally to the XB peptide.

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]

@@ Line 7: / Line 7: @@
 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.
-This construct 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: [2] -the N terminal Core domain (113 AA) -the C terminal XB peptide (25 AA) -and a 'flexible linker' in between the first parts (28 AA) We used the same domain structure as is used within the Rapamycin inducable split system. [1] While the Core domain is responsible for dimerization of sspB and binding of ssrA the XB peptide 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. [3]
+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)
-For our project we needed the sspB split system to engineer a light inducable sspB.
+-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 peptide 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.
 There is no start codon included in this part, because regulatory parts are fused N-terminally to the XB peptide.