Difference between revisions of "Part:BBa K1638005"
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− | 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 | + | 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. |
The rise in cyclic AMP can trigger the expression of genes by using a cAMP-induced promotor. This promotor could for example regulate the expression of a red flourescent protein (RFP). The presence of red-fluorescent cells can in turn be used to verify protein-protein interactions. | The rise in cyclic AMP can trigger the expression of genes by using a cAMP-induced promotor. This promotor could for example regulate the expression of a red flourescent protein (RFP). The presence of red-fluorescent cells can in turn be used to verify protein-protein interactions. | ||
Revision as of 10:41, 11 September 2015
T18 domain of cyaA from Bordetella pertussis (IPTG inducible)
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. The rise in cyclic AMP can trigger the expression of genes by using a cAMP-induced promotor. This promotor could for example regulate the expression of a red flourescent protein (RFP). The presence of red-fluorescent cells can in turn be used to verify protein-protein interactions.
See BBa_K1638003 for the T25 domain.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 711
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 252
Illegal NgoMIV site found at 662
Illegal AgeI site found at 468 - 1000COMPATIBLE WITH RFC[1000]