Difference between revisions of "Part:BBa K4390026"
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==Usage and Biology==
 
==Usage and Biology==
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Metallothionein (MT) is a small protein (around 6-7 kDa) which is rich in cysteine. These thiol group in cysteines provide ability to chelate almost all heavy metal ions including Cd2+, Hg2+, Pb2+ and As3+, but had been shown that has higher binding affinity with Hg2+ (Manceau, A. et al., 2019). The ability of chelating heavy metals provides the metal tolerance for its hosts. This MT sequence was obtained from Mytilus edulis, a blue mussel which originally live in aqueous environment (MACKAY E. A. et al.,1993).
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SB7 is a 7-amino-acid silica-binding motif from the spore coat protein CotB1 of Bacillus cereus. According to studies, its three arginine residues are important for binding to silica surfaces. We tagged this sequence to the N-terminus of the metallothionein of Mytilus edulis. Therefore, when the lysate from a cell culture expressing SB7-tagged metallothionein is mixed with silica beads followed by washing off the supernatant, we would be left with metallothioneins bound to the silica beads. This entire construct could then be applied as an ultrafiltration device for removing heavy metals from the wastewater. This is one of the three types of silica-binding tags we explored for immobilising proteins to silica beads.  
 
SB7 is a 7-amino-acid silica-binding motif from the spore coat protein CotB1 of Bacillus cereus. According to studies, its three arginine residues are important for binding to silica surfaces. We tagged this sequence to the N-terminus of the metallothionein of Mytilus edulis. Therefore, when the lysate from a cell culture expressing SB7-tagged metallothionein is mixed with silica beads followed by washing off the supernatant, we would be left with metallothioneins bound to the silica beads. This entire construct could then be applied as an ultrafiltration device for removing heavy metals from the wastewater. This is one of the three types of silica-binding tags we explored for immobilising proteins to silica beads.  
  
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<partinfo>BBa_K4390026 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K4390026 SequenceAndFeatures</partinfo>
  
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==References==
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MACKAY, E. A. et al. (1993) Complete amino acid sequences of five dimeric and four monomeric forms of metallothionein from the edible mussel Mytilus edulis. European journal of biochemistry. 218 (1), 183–194.
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Manceau, A. et al. (2019) Mercury(II) Binding to Metallothionein in Mytilus edulis revealed by High Energy‐Resolution XANES Spectroscopy. Chemistry : a European journal. 25 (4), 997–1009.
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  

Revision as of 15:43, 13 October 2022


SB7-tagged M. edulis Metallothionein

This part is not compatible with BioBrick RFC10 assembly but is compatible with the iGEM Type IIS Part standard which is also accepted by iGEM.

Usage and Biology

Metallothionein (MT) is a small protein (around 6-7 kDa) which is rich in cysteine. These thiol group in cysteines provide ability to chelate almost all heavy metal ions including Cd2+, Hg2+, Pb2+ and As3+, but had been shown that has higher binding affinity with Hg2+ (Manceau, A. et al., 2019). The ability of chelating heavy metals provides the metal tolerance for its hosts. This MT sequence was obtained from Mytilus edulis, a blue mussel which originally live in aqueous environment (MACKAY E. A. et al.,1993).

SB7 is a 7-amino-acid silica-binding motif from the spore coat protein CotB1 of Bacillus cereus. According to studies, its three arginine residues are important for binding to silica surfaces. We tagged this sequence to the N-terminus of the metallothionein of Mytilus edulis. Therefore, when the lysate from a cell culture expressing SB7-tagged metallothionein is mixed with silica beads followed by washing off the supernatant, we would be left with metallothioneins bound to the silica beads. This entire construct could then be applied as an ultrafiltration device for removing heavy metals from the wastewater. This is one of the three types of silica-binding tags we explored for immobilising proteins to silica beads.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 11
    Illegal NheI site found at 34
    Illegal NotI site found at 73
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

References

MACKAY, E. A. et al. (1993) Complete amino acid sequences of five dimeric and four monomeric forms of metallothionein from the edible mussel Mytilus edulis. European journal of biochemistry. 218 (1), 183–194.

Manceau, A. et al. (2019) Mercury(II) Binding to Metallothionein in Mytilus edulis revealed by High Energy‐Resolution XANES Spectroscopy. Chemistry : a European journal. 25 (4), 997–1009.