Difference between revisions of "Part:BBa K2389010"
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First introduced in 1998, the bacterial two-hybrid system (BACTH) based in Escherichia coli facilitated the screening of interactions between two proteins, and provided a simpler, yet powerful, alternative to the well-known yeast two-hybrid technology. BACTH utilizes the catalytic domain of Bordetella pertussis adenylate cyclase. Here, the two complementary fragments of adenylate cyclase, T18 and T25, are each fused to one of the proteins of interest. Good interaction between the two proteins allows for the reconstitution of the two halves of adenylate cyclase, thus restoring the synthesis of cyclic AMP (cAMP) from ATP (Figure 1). In catabolic operons, such as the lac operon, cyclic AMP binds to the catabolite activator protein (CAP), increasing the affinity of CAP for DNA, and thus transcription through CAP’s interaction with RNA Polymerase. | First introduced in 1998, the bacterial two-hybrid system (BACTH) based in Escherichia coli facilitated the screening of interactions between two proteins, and provided a simpler, yet powerful, alternative to the well-known yeast two-hybrid technology. BACTH utilizes the catalytic domain of Bordetella pertussis adenylate cyclase. Here, the two complementary fragments of adenylate cyclase, T18 and T25, are each fused to one of the proteins of interest. Good interaction between the two proteins allows for the reconstitution of the two halves of adenylate cyclase, thus restoring the synthesis of cyclic AMP (cAMP) from ATP (Figure 1). In catabolic operons, such as the lac operon, cyclic AMP binds to the catabolite activator protein (CAP), increasing the affinity of CAP for DNA, and thus transcription through CAP’s interaction with RNA Polymerase. | ||
| − | <p style="text-align:center;"><img src="https://static.igem.org/mediawiki/2017/1/10/T--UAlberta--T1825.png" width=" | + | <p style="text-align:center;"><img src="https://static.igem.org/mediawiki/2017/1/10/T--UAlberta--T1825.png" width="30"></p> |
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Revision as of 02:24, 2 November 2017
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First introduced in 1998, the bacterial two-hybrid system (BACTH) based in Escherichia coli facilitated the screening of interactions between two proteins, and provided a simpler, yet powerful, alternative to the well-known yeast two-hybrid technology. BACTH utilizes the catalytic domain of Bordetella pertussis adenylate cyclase. Here, the two complementary fragments of adenylate cyclase, T18 and T25, are each fused to one of the proteins of interest. Good interaction between the two proteins allows for the reconstitution of the two halves of adenylate cyclase, thus restoring the synthesis of cyclic AMP (cAMP) from ATP (Figure 1). In catabolic operons, such as the lac operon, cyclic AMP binds to the catabolite activator protein (CAP), increasing the affinity of CAP for DNA, and thus transcription through CAP’s interaction with RNA Polymerase.
<img src="
" width="30">
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 633
Illegal BamHI site found at 1605 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 141
Illegal NgoMIV site found at 551
Illegal AgeI site found at 357
Illegal AgeI site found at 609 - 1000COMPATIBLE WITH RFC[1000]