Difference between revisions of "Part:BBa K823043"

Latest revision as of 05:52, 1 November 2014

alternative sigma factor ecf41

This is an alternative sigma factor from B. lincheniformes regulating e.g. P_ydfG. It is the short version lacking the regulation domain and therefore constitutively on.

Our team used this part in the combination with P_spoIVB/P_sspK,P_ydfG and MazF in the [http://2012.igem.org/Team:LMU-Munich/Germination_Stop#Suicide_switch Suicide switch].

σ G, the last sigma factor in the forespore, activates PspoIVB, producing ECF41 during Sporulation. This then activates the PydfG promoter producing MazF, theoretically killing cells after germination.

This Platereader measurment is the first and not reproduced to this point!

This Platereader assay was done with the Bacillus subtilis strain W168 containing thrC::P_spoIVB-ecf41_{Bli aa 1-204} (through transformation with pSBBs4S-P_spoIVB-ecf41_{Bli aa 1-204}) and sacA::P_ydfG-luxABCDE (through transformation with pSBBs3C-luxABCDE-P_ydfG). That we get a signal shows that this is active together with P_ydfG.

Fig. 1. Platereader measurement of ECF41 delay

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
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 443
1000
COMPATIBLE WITH RFC[1000]

@@ Line 13: / Line 13: @@
 [[Image:PlatereaderPspoIVBecf41PydfG.jpg|600px]]
-This Platereader assay was done with the ''Bacillus subtilis'' strain W168 containing ''thrC''::P<sub>''spoIVB''</sub>-''ecf41<sub>Bli aa  1-204</sub>'' (through transformation with pSBBs4S-P<sub>''spoIVB''</sub>-''ecf41<sub>Bli aa  1-204</sub>'') and ''sacA''::P<sub>''ydfG''</sub>-''luxABCDE'' (through transformation with pSBBs3C-''luxABCDE''-P<sub>''ydfG''</sub>). That we get a signal shows that this is active together with [https://parts.igem.org/wiki/index.php?title=Part:BBa_K823041 PydfG].
+This Platereader assay was done with the ''Bacillus subtilis'' strain W168 containing ''thrC''::P<sub>''spoIVB''</sub>-''ecf41<sub>Bli aa  1-204</sub>'' (through transformation with pSBBs4S-P<sub>''spoIVB''</sub>-''ecf41<sub>Bli aa  1-204</sub>'') and ''sacA''::P<sub>''ydfG''</sub>-''luxABCDE'' (through transformation with pSBBs3C-''luxABCDE''-P<sub>''ydfG''</sub>). That we get a signal shows that this is active together with [https://parts.igem.org/wiki/index.php?title=Part:BBa_K823041 P<sub>''ydfG''</sub>].
-<!-- Add more about the biology of this part here
-===Usage and Biology===
+[[File:delay prezi.png|thumb|600px|center|Fig. 1. Platereader measurement of ECF41 delay]]