Difference between revisions of "Part:BBa K1045002"

Latest revision as of 19:45, 28 October 2013

CFP Reporter under control of the c-di-AMP-dependent YdaO Riboswitch

This part is a reporter system.
It is composed of the ydaO riboswitch from Bacillus subtilis genomic DNA with its native promoter and its native RBS (BBa_K1045004), controlling the expression of cfp as a reporter gene (BBa_E0020).

Ideally, in the presence of c-di-AMP, the riboswitch leads to premature termination of cfp transcription. In the absence of c-di-AMP, the system uses the terminator present in the pSB1C3 backbone. This results in the formation of a cfp mRNA which can be translated in a functional CFP protein. Thus, in the abscence of c-di-AMP, the system should fluoresce. In the presence of c-di-AMP, however, the system should not fluoresce.

Furthermore, in combination with the diadenylate cyclase domain (BBa_K1045003), the riboswitch reporter system could indicate the state of the c-di-AMP biosynthesis. This could be exploited in order to screen substances for effects on c-di-AMP homeostasis.

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 3: / Line 3: @@
 This part is a reporter system.<br />
-It was constructed by amplification of a riboswitch from Bacillus subtilis DNA with a native promoter and native RBS ([[Part:BBa_K1045004|BBa_K1045004]]) and combined to a CFP reporter ([[Part:BBa_E0020|BBa_E0020]]). It works as a sensor of c-di-AMP, which enhances the terminating efficiency in the translation of the RNA by ribosomes. The system uses the terminator present in the '''pSB1C3''' backbone. Altogether in the presence of c-di-AMP the system should be hindered and not be glowing.
+It is composed of the ''ydaO'' riboswitch from ''Bacillus subtilis'' genomic DNA with its native promoter and its native RBS ([[Part:BBa_K1045004|BBa_K1045004]]), controlling the expression of ''cfp'' as a reporter gene ([[Part:BBa_E0020|BBa_E0020]]).
-Furthermore, in combination with the diadenylate cyclase domain (like a further recombined part [[Part:BBa_K1045003|BBa_K1045003]]) it might show further interaction. The diadenylate cyclase domain can have the competence to likewise hinder the system, which turns out with no illumination.
+Ideally, in the presence of c-di-AMP, the riboswitch leads to premature termination of ''cfp'' transcription. In the absence of c-di-AMP, the system uses the terminator present in the '''pSB1C3''' backbone. This results in the formation of a ''cfp'' mRNA which can be translated in a functional CFP protein. Thus, in the abscence of c-di-AMP, the system should fluoresce. In the presence of c-di-AMP, however, the system should not fluoresce.
+Furthermore, in combination with the diadenylate cyclase domain ([[Part:BBa_K1045003|BBa_K1045003]]), the riboswitch reporter system could indicate the state of the c-di-AMP biosynthesis. This could be exploited in order to screen substances for effects on c-di-AMP homeostasis.
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