Difference between revisions of "Part:BBa K343003"

Revision as of 11:52, 11 October 2010

NpSopII-NpHtrII-StTar (M-fusion)

Sensory Rhodopsin II bluelight receptor fused to its transducer, HtrII, with a 27 BP linker region. This is fused to Salmonella enterica serovar typhimurium chemotaxis protein Tar, so that the protein effectively couples the input from the receptor to the chemotaxis pathway. The fusion between HtrII and Tsr is an M-fusion in the HAMP domain, which is supposed to give it maximum activity. Sequencing confirmed that the sequence is identical to the found in the article by Jung, Spudich E, Trivedi and Spudich J in the article: An Archaeal Photosignal-Transducing Module Mediates Phototaxis in Escherichia coli. (1)

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NotI site found at 1783
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 73
Illegal NgoMIV site found at 331
Illegal AgeI site found at 1585
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 1027
Illegal BsaI.rc site found at 1300
Illegal SapI site found at 801
Illegal SapI.rc site found at 1801

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 <partinfo>BBa_K343003 short</partinfo>
-Sensory Rhodopsin II bluelight receptor fused to its transducer, HtrII, with a 27 BP linker region.  This is fused with E.Coli's chemotaxis protein Tsr, so that the protein effectively couples the input from the receptor to chemotaxis. The fusion between HtrII and Tsr is an M-fusion in the HAMP domain, which is supposed to give it maximum activity. We deduced the sequence from the instructions given by Jung, Spudich E, Trivedi and Spudich J in the article: An Archaeal Photosignal-Transducing Module Mediates Phototaxis in Escherichia coli. (1)
+Sensory Rhodopsin II bluelight receptor fused to its transducer, HtrII, with a 27 BP linker region.  This is fused to Salmonella enterica serovar typhimurium chemotaxis protein Tar, so that the protein effectively couples the input from the receptor to the chemotaxis pathway. The fusion between HtrII and Tsr is an M-fusion in the HAMP domain, which is supposed to give it maximum activity. Sequencing confirmed that the sequence is identical to the found in the article by Jung, Spudich E, Trivedi and Spudich J in the article: An Archaeal Photosignal-Transducing Module Mediates Phototaxis in Escherichia coli. (1)
 <!-- Add more about the biology of this part here
 ===Usage and Biology===
+When exposed to bluelight, the sensory rhodopsin II will undergo a change in ultrastructure, which is being transduced through HtrII on to the Tar domain. This influences the cell's normal chemotaxis pathway, so that it will decrease the amount of phosphorylated CheY and thereby decrease the tumbling frequency of the cell.
+The part requires retinal to work in E.Coli. This can be achieved through adding retinal to the liquid growth medium and/or the plates. Currently we are doing experiments on wether the part also functions with an internal retinal source, ie retinal synthesis in E. Coli.
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