Difference between revisions of "Part:BBa K1638032"

Revision as of 00:15, 18 September 2015

T18 domain of cyaA with cAMP-induced RFP generator

This part contains the T25 domain of the adenylate cyclase cyaA from Bordetella pertussis. This part is intented to be used in the bacterial two-hybrid system. This system is based on the reconstitution of the adenylate cyclase. When protein-coding genes is suffixed to the T25 and T18 domains of cyaA, one can study the interaction of these two proteins. If the conjugated proteins interacts, T25 and T18 will be brought into close proximity. This will enable the two catalytic domains T18 and T25 to synthesize cyclic adenosinemonophosphate (cAMP). The rise in cAMP can trigger the expression of genes by using a cAMP-induced promotor that induce the transcription of red fluorescent protein (RFP). For this reason this part also contains a mRFP generator controlled by PcstA BBa_K861173. The presence of red-fluorescent cells can in turn be used to verify protein-protein interactions.

See BBa_K1638033 for the T25 domain.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 1719
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 1260
Illegal NgoMIV site found at 1670
Illegal AgeI site found at 712
Illegal AgeI site found at 824
Illegal AgeI site found at 1476
1000
COMPATIBLE WITH RFC[1000]

@@ Line 2: / Line 2: @@
 <partinfo>BBa_K1638032 short</partinfo>
-The T18 domain of CyaA to be used in the bacterial two-hybrid system. The expression of the gene is under control of a lac operator and is induced by Isopropyl β-D-1-thiogalactopyranoside (IPTG). When protein-coding genes is suffixed to the T25 and T18 domains of the bacterial two-hybrid system, the interaction of these two proteins can be examined. If the conjugated proteins associates, T25 and T18 associates too. This leads to the catalysation of the conversion of ATP to cAMP.
+This part contains the T25 domain of the adenylate cyclase <i>cyaA</i> from <i>Bordetella pertussis</i>. This part is intented to be used in the bacterial two-hybrid system. This system is based on the reconstitution of the adenylate cyclase. When protein-coding genes is suffixed to the T25 and T18 domains of <i>cyaA</i>, one can study the interaction of these two proteins. If the conjugated proteins interacts, T25 and T18 will be brought into close proximity. This will enable the two catalytic domains T18 and T25 to synthesize cyclic adenosinemonophosphate (cAMP).
-The rise in cyclic AMP can trigger the expression of genes by using a cAMP-induced promotor. For this construct a reporter system based on RFP controlled by the promoter PcstA is used. The presence of red-fluorescent cells can be used to verify protein-protein interactions.
+The rise in cAMP can trigger the expression of genes by using a cAMP-induced promotor that induce the transcription of red fluorescent protein (RFP). For this reason this part also contains a mRFP generator controlled by PcstA <partinfo>BBa_K861173</partinfo>. The presence of red-fluorescent cells can in turn be used to verify protein-protein interactions.
 See <partinfo>BBa_K1638033</partinfo> for the T25 domain.