Difference between revisions of "Part:BBa K590015"
(→‎Usage and Biology)
 
(9 intermediate revisions by 2 users not shown)
Line 5: Line 5:
  
 
===Usage and Biology===
 
===Usage and Biology===
−
This part consists of mamK gene from ''Magnetospirillum magneticum'' strain AMB-1, sfGFP in the backbone of [https://parts.igem.org/wiki/index.php?title=Part:BBa_K590014 pGA3k3]. It was made by Gibson Cloning.
 
  
−
mamK was previously reported for its importance proper magnetosome chain organization in magnetic bacteria; it is bacterial actin-like cytoskeleton protein required for proper alignment of the magnetosomes in a chain.  
+
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 filament fluorescent structures were observed:
+
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.
  
−
[[File:MamK-1 uwigem2011.jpg]]
+
[[Image:MamK sfGFP 3k3-1 uwigem2011part.jpg|300px|center]]
 +
<html><center>
 +
<iframe width="420" height="315" src="http://www.youtube.com/embed/BLLyUHrrcV4" frameborder="0" allowfullscreen></iframe>
 +
</center></html>
  
 
<!-- -->
 
<!-- -->

Latest revision as of 18:20, 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.

MamK sfGFP 3k3-1 uwigem2011part.jpg

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