Difference between revisions of "Part:BBa K590015"
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Revision as of 18:17, 31 October 2011
sfGFP_mamK_pGA1C3
This part was made by [http://2011.igem.org/Team:Washington UW iGEM Team 2011] and was contributed to the [http://2011.igem.org/Team:Washington/Magnetosomes/Results Magnetosome Toolkit].
Usage and Biology
This part consists of the mamK gene from Magnetospirillum magneticum strain AMB-1, fused to superfolder gfp on pGA1C3. MamK has been reported to be essential for proper magnetosome formation in magnetotactic bacteria; its bacterial actin-like cytoskeleton protein is required for proper alignment of the magnetosomes in a chain. Beyond magnetosome alignment, MamK filaments could act as a scaffold for protein localization or to alter cell morphology by preventing cells from fully dividing (see images below).
We transformed this part into an E. coli BL-21 lacIq strain and observed fluorescent filament structures that in some cases, prevented cells from dividing properly. The same construct has been successfully cloned into Gibson assembly vectors pGA1A3 and pGA4C5.
<iframe width="420" height="315" src="http://www.youtube.com/embed/BLLyUHrrcV4" frameborder="0" allowfullscreen></iframe>
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 1430
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 1430
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1134
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 1430
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 1430
Illegal NgoMIV site found at 1156
Illegal NgoMIV site found at 1523
Illegal NgoMIV site found at 1670 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 80
Illegal SapI.rc site found at 1802