Difference between revisions of "Part:BBa K3165000"
| (2 intermediate revisions by 2 users not shown) | |||
| Line 3: | Line 3: | ||
<partinfo>BBa_K3165000 short</partinfo> | <partinfo>BBa_K3165000 short</partinfo> | ||
| − | A miniaturized CcaS variant which lacks the two PAS domains, due to deletion of the segment following the GAF domain until the linker before the Histidine Kinase domain. Mini-CcaS as a green light-sensitive system has a much higher dynamic fold change (around 600) as compared to the | + | A miniaturized CcaS variant which lacks the two PAS domains, due to deletion of the segment following the GAF domain until the linker before the Histidine Kinase domain. Mini-CcaS as a green light-sensitive system has a much higher dynamic fold change (around 600) as compared to the wild type found naturally in <i>Synechocystis</i>. |
| Line 14: | Line 14: | ||
===Usage and Biology=== | ===Usage and Biology=== | ||
<h2> Biology </h2> | <h2> Biology </h2> | ||
| − | CcaSR is a green-light activated two-component signal transduction system (TCS) that controls the expression of phycobilisome linker protein CpcG2 in <i>Synechocystis</i> PCC6803 complementary chromatic adaptation pathway. The naturally occurring CcaS protein is a cyanobacteriochrome sensor histidine kinase consisting of an N-terminal GAF domain (cGMP phosphodiesterase/ adenyl cyclase/ FhlA) bound to two PAS domains of unknown function attached to a C-terminal Histidine Kinase domain. Upon activation with green light, the CcaS switches to a red-absorbing conformation that phosphorylates the response regulator CcaR leading to the activation of the PcpcG2 promoter initiating transcription. On illumination with green light, a 6-fold activation of the protein was reported in the wild type protein. | + | CcaSR is a green-light activated two-component signal transduction system (TCS) that controls the expression of phycobilisome linker protein CpcG2 in <i>Synechocystis</i> sp. PCC6803 complementary chromatic adaptation pathway. The naturally occurring CcaS protein is a cyanobacteriochrome sensor histidine kinase consisting of an N-terminal GAF domain (cGMP phosphodiesterase/ adenyl cyclase/ FhlA) bound to two PAS domains of unknown function attached to a C-terminal Histidine Kinase domain. Upon activation with green light, the CcaS switches to a red-absorbing conformation that phosphorylates the response regulator CcaR leading to the activation of the PcpcG2 promoter initiating transcription. On illumination with green light, a 6-fold activation of the protein was reported in the wild type protein. |
Mini-CcaS is a modified form of the naturally occurring CcaS protein which lacks the two PAS domains. The modified protein has highly reduced leakiness and a fold change of around 593, qualifying as a much more sensitive optogenetic system. | Mini-CcaS is a modified form of the naturally occurring CcaS protein which lacks the two PAS domains. The modified protein has highly reduced leakiness and a fold change of around 593, qualifying as a much more sensitive optogenetic system. | ||
<h2> Usage </h2> | <h2> Usage </h2> | ||
| − | This part can be used to optically regulate the expression of the gene of interest downstream of the | + | This part can be used to optically regulate the expression of the gene of interest downstream of the PcpcG2 promoter upon stimulation with the appropriate frequency of light. This part in association with the response regulator protein - CcaR can be used as a genetic circuit for optically programmed expression signals. |
<h3> References :</h3> | <h3> References :</h3> | ||
Latest revision as of 18:48, 21 October 2019
mini-CcaS
A miniaturized CcaS variant which lacks the two PAS domains, due to deletion of the segment following the GAF domain until the linker before the Histidine Kinase domain. Mini-CcaS as a green light-sensitive system has a much higher dynamic fold change (around 600) as compared to the wild type found naturally in Synechocystis.
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]
Usage and Biology
Biology
CcaSR is a green-light activated two-component signal transduction system (TCS) that controls the expression of phycobilisome linker protein CpcG2 in Synechocystis sp. PCC6803 complementary chromatic adaptation pathway. The naturally occurring CcaS protein is a cyanobacteriochrome sensor histidine kinase consisting of an N-terminal GAF domain (cGMP phosphodiesterase/ adenyl cyclase/ FhlA) bound to two PAS domains of unknown function attached to a C-terminal Histidine Kinase domain. Upon activation with green light, the CcaS switches to a red-absorbing conformation that phosphorylates the response regulator CcaR leading to the activation of the PcpcG2 promoter initiating transcription. On illumination with green light, a 6-fold activation of the protein was reported in the wild type protein. Mini-CcaS is a modified form of the naturally occurring CcaS protein which lacks the two PAS domains. The modified protein has highly reduced leakiness and a fold change of around 593, qualifying as a much more sensitive optogenetic system.
Usage
This part can be used to optically regulate the expression of the gene of interest downstream of the PcpcG2 promoter upon stimulation with the appropriate frequency of light. This part in association with the response regulator protein - CcaR can be used as a genetic circuit for optically programmed expression signals.
References :
(1) A miniaturized E. coli green light sensor with a high dynamic range. Ong NTX, Tabor JJ. Chembiochem. 2018 Feb 8. doi: 10.1002/cbic.201800007. 10.1002/cbic.201800007 PubMed 29420866
(2) M. Nakajima, S. Ferri, M. Rögner, K. Sode, Sci. Rep. 2016, 6, 37595
(3) Y. Hirose, R. Narikawa, M. Katayama, M. Ikeuchi, Proc. Natl. Acad. Sci. U. S. A.
2010, 107, 8854–8859.