Part:BBa_K590016

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 lacI^q strain and observed strong membrane localization.

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.

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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