Difference between revisions of "Part:BBa K343003:Design"
(→Design Notes) |
(→Source) |
||
| Line 14: | Line 14: | ||
===Source=== | ===Source=== | ||
| − | + | Sensory Rhodopsin II and HtrII domains were taken from the genome of Natronomonas Pharaonis. The Tar methyl accepting chemotaxis protein was taken from Salmonella Enterica Serovar Typhimurium. The synthetic linker and fusing of the HAMP domains of HtrII and StTar was done by the team behind the referenced article (1). | |
| − | + | ||
===References=== | ===References=== | ||
[http://jb.asm.org/cgi/content/full/183/21/6365] Jung K-H, Spudich EN, Trivedi VD and Spudich JL ; An Archaeal Photosignal-Transducing Module Mediates Phototaxis in Escherichia coli; Journal of Bacteriology, Nov. 2001, p. 6365–6371. | [http://jb.asm.org/cgi/content/full/183/21/6365] Jung K-H, Spudich EN, Trivedi VD and Spudich JL ; An Archaeal Photosignal-Transducing Module Mediates Phototaxis in Escherichia coli; Journal of Bacteriology, Nov. 2001, p. 6365–6371. | ||
Revision as of 09:04, 13 October 2010
NpSopII-NpHtrII-StTar (M-fusion)
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 1783
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 73
Illegal NgoMIV site found at 331
Illegal AgeI site found at 1585 - 1000INCOMPATIBLE 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
Design Notes
NpSopII-NpHtrII-StTar fusion-chimera protein sequence from Jung K-H, Spudich EN, Trivedi VD and Spudich JL (1).
The first 224 amino acid residues come from the NpSopII gene, encoding a bluelight photon receptor with 15 residues removed at the C-terminal end. The following 9 amino acids are a linker. the last part is HtrII fused with Tsr from E.Coli at the HAMP domain. The complex' first 125 amino acid residues come from HtrII and the remaining 279 from Tsr.
The constitutive promoter R0040 was chosen for its medium strength, so that there would be a rather large amount of the fusion, chimera protein expressed, but not so much that it would be lethal to the cell. The easy repression and inhibition of repression through TetR and tetracycline makes for an easily controlled expression if that is needed. The doible terminator B0015 was chosen for it's reliability and good reviews on the parts.igem.
Source
Sensory Rhodopsin II and HtrII domains were taken from the genome of Natronomonas Pharaonis. The Tar methyl accepting chemotaxis protein was taken from Salmonella Enterica Serovar Typhimurium. The synthetic linker and fusing of the HAMP domains of HtrII and StTar was done by the team behind the referenced article (1).
References
[http://jb.asm.org/cgi/content/full/183/21/6365] Jung K-H, Spudich EN, Trivedi VD and Spudich JL ; An Archaeal Photosignal-Transducing Module Mediates Phototaxis in Escherichia coli; Journal of Bacteriology, Nov. 2001, p. 6365–6371.