Difference between revisions of "Part:BBa K2042006"
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<partinfo>BBa_K2042006 short</partinfo> | <partinfo>BBa_K2042006 short</partinfo> | ||
− | This is a sequence for a cyclohexanone inductible promoter. This is a promoter specially adapted for the use in | + | This is a sequence for a cyclohexanone inductible promoter. This is a promoter specially adapted for the use in gram negative bacteria [1], and Pseudomonas putida is a very good candidate because it is able to thrive in environments containing haloalkanes and other cyclic carbon compounds. |
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+ | This promoter is based on elements of the cyclohexanol biodegradation pathway of Acinetobacter johnsonii.edited in silico to obtain a standardized SEVA (Standard European Vector Architecture) expression cargo, and assembled to yield plasmid pSEVA2311. | ||
+ | It was edited and formatted from ChnR/PchnB regulatory node of Acinetobacter johnsonii to ease the targeted engineering of ectopic gene expression in Gram-negative bacteria. The system was assembled in a minimal DNA segment adopting a Synthetic Biology standard. | ||
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+ | Transcriptional activation of the ChnR/PchnB expression system upon addition of cyclohexanone was evaluated in a wild-type Escherichia coli strain. The expression data presented reflect a tightly controlled transcription initiation signal in response to cyclohexanone. | ||
+ | Key features of this expression vector include [i] a very low expression level in the absence of inducer, [ii] high transcriptional capacity, [iii] an induction kinetics very similar in both minimal and rich culture media, and [iv] linear accumulation of the reporter product along time | ||
+ | |||
+ | According to the literature a concentrations of inducer (i.e. cyclohexanone) of 1 mM is suitable for gene expression in gram negative bacteria [1]. | ||
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− | [1] Benedetti, I. | + | [1] Benedetti, Ilaria, Pablo I. Nikel, and Víctor de Lorenzo. "Data on the standardization of a cyclohexanone-responsive expression system for Gram-negative bacteria." Data in brief 6 (2016): 738-744. |
Revision as of 23:20, 29 October 2016
Cyclohexanone inducible promoter
This is a sequence for a cyclohexanone inductible promoter. This is a promoter specially adapted for the use in gram negative bacteria [1], and Pseudomonas putida is a very good candidate because it is able to thrive in environments containing haloalkanes and other cyclic carbon compounds.
This promoter is based on elements of the cyclohexanol biodegradation pathway of Acinetobacter johnsonii.edited in silico to obtain a standardized SEVA (Standard European Vector Architecture) expression cargo, and assembled to yield plasmid pSEVA2311. It was edited and formatted from ChnR/PchnB regulatory node of Acinetobacter johnsonii to ease the targeted engineering of ectopic gene expression in Gram-negative bacteria. The system was assembled in a minimal DNA segment adopting a Synthetic Biology standard.
Transcriptional activation of the ChnR/PchnB expression system upon addition of cyclohexanone was evaluated in a wild-type Escherichia coli strain. The expression data presented reflect a tightly controlled transcription initiation signal in response to cyclohexanone. Key features of this expression vector include [i] a very low expression level in the absence of inducer, [ii] high transcriptional capacity, [iii] an induction kinetics very similar in both minimal and rich culture media, and [iv] linear accumulation of the reporter product along time
According to the literature a concentrations of inducer (i.e. cyclohexanone) of 1 mM is suitable for gene expression in gram negative bacteria [1].
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
[1] Benedetti, Ilaria, Pablo I. Nikel, and Víctor de Lorenzo. "Data on the standardization of a cyclohexanone-responsive expression system for Gram-negative bacteria." Data in brief 6 (2016): 738-744.