Difference between revisions of "Part:BBa K590015"

Revision as of 20:18, 28 September 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 lacI^q 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.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal PstI site found at 1430
12
INCOMPATIBLE WITH RFC[12]
Illegal PstI site found at 1430
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 1134
23
INCOMPATIBLE WITH RFC[23]
Illegal PstI site found at 1430
25
INCOMPATIBLE 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
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI.rc site found at 80
Illegal SapI.rc site found at 1802

@@ Line 8: / Line 8: @@
 This part consists of the mamK gene from Magnetospirillum magneticum strain AMB-1, fused to superfolder gfp on [https://parts.igem.org/wiki/index.php?title=Part:BBa_K590011 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).
-This part was transformed into ''E. coli'' and fluorescent filament structures were observed. The same construct has been successfully cloned into Gibson assembly vectors pGA1A3 and pGA4C5.
+We transformed this part into an ''E. coli'' BL-21 lacI<sup>q</sup> 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.
 [[Image:MamK sfGFP 3k3-1 uwigem2011part.jpg|500px|center]]