Team:Washington/Magnetosomes/Parts

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iGEM Toolkits: Parts Submitted


The iGEM toolkits group submitted a total twenty-two parts to the registry:

  • 10 magnetosome gene groups from mamAB operon
  • 3 superassemblies of the mamAB operon from the set of 10 gene groups
  • 2 sfGFP-mamK and sfGFP-mamI gene fusions
  • 5 Gibson Assembly (pGA) vectors

Magnetosome Toolkit Favorite Parts

[http://partsregistry.org/wiki/index.php?title=Part:BBa_K590015 BBa_K590015 sfGFP_mamK_pGA1C3]

This part consists of the mamK gene from Magnetospirillum magneticum strain AMB-1, fused to superfolder gfp on pGA1C3. MamK was previously reported to be essential for proper magnetosome chain alignment in magnetotactic bacteria; its bacterial actin-like cytoskeleton protein is required for proper alignment of the magnetosomes in a chain. Beyond magnetosome alignment, its filament could act as a scaffold for protein localization or to alter cell morphology.

[http://partsregistry.org/wiki/index.php?title=Part:BBa_K590016 BBa_K590016 sfGFP_mamI_pGA1C3]

This part consists of the mamI gene from Magnetospirillum magneticum strain AMB-1, fused to superfolder gfp on pGA1C3. MamI is a membrane-localized protein that is essential for magnetosome vesicle formation, and is known to also bind the MamK filament.

The Gibson Assembly Toolkit

[http://partsregistry.org/wiki/index.php?title=Part:BBa_K590010 BBa_K590010 pGA1A3_pLacGFP]

This is a high copy plasmid backbone (replication origin pMB1) that confers ampicillin resistance. It has an insert with a LacI-repressible promoter driving GFP expression. It is identical to pSB1A3 except for its bglBrick prefix and suffix, and is optimized for Gibson Assembly.

[http://partsregistry.org/wiki/index.php?title=Part:BBa_K590011 BBa_K590011 pGA1C3_pLacGFP]

This is a high copy plasmid backbone (replication origin pMB1) that confers chloramphenicol resistance. It has an insert with a LacI-repressible promoter driving GFP expression. It is identical to pSB1C3 except for its bglBrick prefix and suffix, and is optimized for Gibson Assembly.

[http://partsregistry.org/wiki/index.php?title=Part:BBa_K590014 BBa_K590014 pGA3K3_pLacGFP]

This is a medium copy plasmid backbone (replication origin p15A) that confers kanamycin resistance. It has an insert with a LacI-repressible promoter driving GFP expression. pGA3K3 is identical to pSB3K3 except for its bglBrick prefix and suffix, and is optimized for Gibson Assembly.

[http://partsregistry.org/wiki/index.php?title=Part:BBa_K590012 BBa_K590012 pGA4C5_pLacGFP]

This is a low copy plasmid backbone (replication origin pSC101) that confers chloramphenicol resistance. It has an insert with a LacI-repressible promoter driving GFP expression. It is identical to pSB4C5 except for its bglBrick prefix and suffix, and is optimized for Gibson Assembly.

[http://partsregistry.org/wiki/index.php?title=Part:BBa_K590013 BBa_K590013 pGA4A5_pLacGFP]

This is a low copy plasmid backbone (replication origin pSC101) that confers ampicillin resistance. It has an insert with a LacI-repressible promoter driving GFP expression. It is identical to pSB4A5 except for its bglBrick prefix and suffix, and is optimized for Gibson Assembly. It is also notable that though we searched in multiple BioBrick distributions from different years, we could not find any that could be sequence-verified as pSB4A5 (the Vf2 sequence reads all suggested pSB1A2). We then generated pGA4A5 from pGA4C5 using the ampicillin cassette from pSB1A3.

Gene AMB Number Cluster Membership Member of 28 genes list? (specific*/related**) Function Summary (Vesicle chain formation, and/or biomineralization) Gene Function
mamH amb0961 mamAB Related
mamI amb0962 mamAB Specific Vesicle, (Chain Formation?) >berkeley 2010: Loss causes no membrane formation, is localized onto chains
mamE amb0963 mamAB; mam Islet Related >Membrane-bound serine protease required for magnetite formation; might control the localization of other magnetosome proteins
mamJ amb0964 mamAB; mam Islet Specific Chain Formation >Proper magnetosome chain organization/assembly
mamK amb0965 mamAB; mam Islet Related Chain Formation >required for proper magnetosome chain organization; *bacterial actin-like cytoskeleton protein required for proper alignment of the magnetosomes in a chain, shown to localize the mamI
mamL amb0966 mamAB; mam Islet Specific Vesicle, biomineralization >berkely 2010: Crucial to mangneosome membrane creation, shown to be spread across the cell membrane and sometimes forms lines
mamM amb0967 mamAB Related >biomineralization, involved in iron transport, magnetite nucleation, or establishement of the proper chemical enviornment for magnetite synthesis in the magnetosome
mamN amb0968 mamAB Related >biomineralization, involved in iron transport, magnetite nucleation, or establishement of the proper chemical enviornment for magnetite synthesis in the magnetosome
mamO amb0969 mamAB Related >biomineralization, involved in iron transport, magnetite nucleation, or establishement of the proper chemical enviornment for magnetite synthesis in the magnetosome
mamP amb0970 mamAB Related Biomineralization >berkeley 2010: loss causes weak magnetic response, with large but fewer crystals
mamA amb0971 mamAB Related >Required for magnetosome activation; activation of vessicles
mamQ amb0972 mamAB; mam Islet Related >ORF; formation/maintenance of magnetosome membranes
mamR amb0973 mamAB Specific Chain formation, Biomineralization >ORF; plays a role in controlling both particle number and size of magnetite cyrstals
mamB amb0974 mamAB Related Vesicle, Biomineralization >indirect role in magnetosome membrane invagination and biomineralization; magnetosome compartment formation
mamS amb0975 mamAB Specific
mamT amb0976 mamAB Specific Biomineralization >magnetite crystal growth; participates in different steps during magnetite synthesis
mamU amb0977 mamAB Related
mamV amb0978 mamAB N/A