Difference between revisions of "Part:BBa K1985000"

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<partinfo>BBa_K1985000 short</partinfo>
 
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The mamP gene from the magnetosome operon mamAB forms a protein dimer. It is an iron-oxidase that contributes to the formation of magnetite by mediating one step in the process, the formation of iron(III)ferrihydrite.
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===Usage and Biology===
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Usage: The mamP gene produces a protein that can be used to transfer electrons to iron molecules. It can be used in combination with other mam genes to form an electron transport complex (reference) that should form magnetic magnetite crystals when the cell is exposed to iron in solution. It was to assemble a construct of MamO, P, T and X in pSB1A3 for in vivo expression. 
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Biology:
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MamP is a proposed cytochrome protein (reference) due to the presence of a heme binding motif within its sequence. It forms a complex with other Mam proteins on the membrane of the native species, Magnetospirillum gryphiswaldense. Together they promote magnetite crystal maturation within an organelle called the magnetosome.
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===References===
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Siponen, Marina I. et al. "Structural Insight Into Magnetochrome-Mediated Magnetite Biomineralization". Nature 502.7473 (2013): 681-684. Web. 14 Oct. 2016.
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<partinfo>BBa_K1985000 parameters</partinfo>
 
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===Usage and Biology===
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Usage: The mamP gene produces a protein that can be used to transfer electrons to iron molecules, specfically, in vitro it can form iron(III)ferrihydrite. It can be used in combination with other mam genes to form an electron transport complex that should form  magnetite (Fe(II)Fe(III)2O4) crystals when the cell is exposed to iron in solution. It was used to assemble a construct of MamO, P, T and X in pSB1A3 for in vivo expression. 
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Biology:
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MamP has been described as a cytochrome protein due to the presence two c-type cytochrome domains, with haem binding domains. It forms a complex with other Mam proteins on the membrane of the native species, Magnetospirillum gryphiswaldense. Together they promote magnetite crystal maturation within an organelle called the magnetosome.
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===Validation===
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The part was first validated with a diagnostic restriction digest using EcoRI and PstI and agarose gel electrophoresis. The expected band sizes from the digest were: 2029 for the plasmid backbone and 871 for the insert. A 1kB plus DNA marker was used to verify the sizes of the bands and it was confirmed that the correct plasmid had been produced.
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[[File:PSB1C3-MamP gel.jpeg||400px|thumb|centre|Figure 2. Agarose gel of the restriction digest of BBa_K1985000 in pSB1C3, with EcoRI and PstI.]]
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===References===
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Siponen, Marina I. et al. "Structural Insight Into Magnetochrome-Mediated Magnetite Biomineralization". Nature 502.7473 (2013): 681-684.

Latest revision as of 12:56, 17 October 2016

mamP

The mamP gene from the magnetosome operon mamAB forms a protein dimer. It is an iron-oxidase that contributes to the formation of magnetite by mediating one step in the process, the formation of iron(III)ferrihydrite.


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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 493
  • 1000
    COMPATIBLE WITH RFC[1000]


Usage and Biology

Usage: The mamP gene produces a protein that can be used to transfer electrons to iron molecules, specfically, in vitro it can form iron(III)ferrihydrite. It can be used in combination with other mam genes to form an electron transport complex that should form magnetite (Fe(II)Fe(III)2O4) crystals when the cell is exposed to iron in solution. It was used to assemble a construct of MamO, P, T and X in pSB1A3 for in vivo expression.

Biology: MamP has been described as a cytochrome protein due to the presence two c-type cytochrome domains, with haem binding domains. It forms a complex with other Mam proteins on the membrane of the native species, Magnetospirillum gryphiswaldense. Together they promote magnetite crystal maturation within an organelle called the magnetosome.

Validation

The part was first validated with a diagnostic restriction digest using EcoRI and PstI and agarose gel electrophoresis. The expected band sizes from the digest were: 2029 for the plasmid backbone and 871 for the insert. A 1kB plus DNA marker was used to verify the sizes of the bands and it was confirmed that the correct plasmid had been produced.

Figure 2. Agarose gel of the restriction digest of BBa_K1985000 in pSB1C3, with EcoRI and PstI.

References

Siponen, Marina I. et al. "Structural Insight Into Magnetochrome-Mediated Magnetite Biomineralization". Nature 502.7473 (2013): 681-684.