Difference between revisions of "Part:BBa K590016"

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===Usage and Biology===
 
===Usage and Biology===
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This part consists of mamI gene from ''Magnetospirillum magneticum'' strain AMB-1, sfGFP in the backbone of [https://parts.igem.org/wiki/index.php?title=Part:BBa_K590011 pGA1C3]. It was made by Gibson Cloning Method.  
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This part consists of the <i>mamI</i> gene from <i>Magnetospirillum magneticum</i> strain AMB-1, fused to  superfolder <i>gfp</i> cloned into [https://parts.igem.org/wiki/index.php?title=Part:BBa_K590011 pGA1C3]. MamI is a membrane-localized protein that is essential for magnetosome vesicle formation, and is also known to bind the [https://parts.igem.org/wiki/index.php?title=Part:BBa_K590015 MamK] filament. Since it localizes to the membrane, MamI may be useful in maximizing the flux of biosynthetic pathways as a membrane-linker protein.
  
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MamI is a membrane-localized protein that localizes magnetic vesicles to the surface of cells, thus forming characteristic magnetosome chains.  It was previously reported that the loss of mamI causes no membrane vesicle formation.
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We transformed this part into a ''E. coli'' BL-21 lacI<sup>q</sup> strain and observed strong membrane localization.
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This part was transformed into ''E.coli'' and membrane localization (particularly the poles) was observed:
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[[Image:SfGFP-I-1C3- uwigem2011.jpg|600px|center]]
 
[[Image:SfGFP-I-1C3- uwigem2011.jpg|600px|center]]
  
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==Contribution==
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Group: Pittsburgh 2019
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Authors: Harrison Green, Victor So, Jemy Varghese, Ripal Sheth, Mel Marciesky
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Summary: In this experiment, the concentration of IPTG was varied to test for optimal expression of the mamI-sfGFP fusion protein. Protein expression was characterized through detection of the fluorescent protein sfGFP. The fluorescent data was normalized using OD600 readings. As demonstrated by the figure below, IPTG concentrations showed minimal effect on expression of mamI-sfGFP protein.
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[[File:T--Pittsburgh--BBa_K590016_characterization_2.svg]]
  
 
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Latest revision as of 09:51, 20 October 2019

sfGFP_mamI_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 mamI gene from Magnetospirillum magneticum strain AMB-1, fused to superfolder gfp cloned into pGA1C3. MamI is a membrane-localized protein that is essential for magnetosome vesicle formation, and is also known to bind the MamK filament. Since it localizes to the membrane, MamI may be useful in maximizing the flux of biosynthetic pathways as a membrane-linker protein.

We transformed this part into a E. coli BL-21 lacIq strain and observed strong membrane localization.

SfGFP-I-1C3- uwigem2011.jpg

Contribution

Group: Pittsburgh 2019

Authors: Harrison Green, Victor So, Jemy Varghese, Ripal Sheth, Mel Marciesky

Summary: In this experiment, the concentration of IPTG was varied to test for optimal expression of the mamI-sfGFP fusion protein. Protein expression was characterized through detection of the fluorescent protein sfGFP. The fluorescent data was normalized using OD600 readings. As demonstrated by the figure below, IPTG concentrations showed minimal effect on expression of mamI-sfGFP protein.

Error creating thumbnail: File missing

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
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 80