Difference between revisions of "Part:BBa K2016003"

Revision as of 09:33, 18 October 2016

McHr - iron-responsive, oxygen-responsive hemerythrin protein coding gene from Methylococcus capsula

McHr is a protein expressed by methane-oxidising bacterium: Methylococcus capsulatus. The family of hemerythrins can undergo a slow process of oxygenation upon binding of iron which causes their colour change from colourless into red/yellow. Its mechanism of oxygenation has been discovered to be much more rapid than other hemerythrins we investigated (DcrHr or TdHr). This gene was synthesised based on the GeneBank sequence entry and cloned into the pSB1C3 plasmid.

Usage and Biology

McHr was used by Sheffield 2016 team to investigate its ability to change colour upon binding of iron and subsequent oxygenation. We cloned our gene into two different plasmids, one under control of strong constitutive promoter and the other under IPTG inducible promoter. We experienced troubles expressing the protein with strong promoters, it is possible that the protein can be toxic to the cell in high concentrations therefore we would suggest using weaker promoters.Biobrick is cloned between XbaI and SpeI sites, however it is cloned in reverse orientation due to XbaI and SpeI enzymes having the same overhangs after restriction digest. To use this biobrick future teams should use PCR apmlification rather than restriction digest to clone this biobrick. Read more about Mc.

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
COMPATIBLE WITH RFC[1000]

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 ===Usage and Biology===
-McHr was used by Sheffield 2016 team to investigate its ability to change colour upon binding of iron and subsequent oxygenation. We cloned our gene into two different plasmids, one under control of strong constitutive promoter and the other under IPTG inducible promoter. We experienced troubles expressing the protein with strong promoters, it is possible that the protein can be toxic to the cell in high concentrations therefore we would suggest using weaker promoters.Biobrick is cloned between XbaI and SpeI sites, however it is cloned in reverse orientation due to XbaI and SpeI enzymes having the same overhangs after restriction digest. To use this biobrick future teams should use PCR apmlification rather than restriction digest to clone this biobrick.
+McHr was used by Sheffield 2016 team to investigate its ability to change colour upon binding of iron and subsequent oxygenation. We cloned our gene into two different plasmids, one under control of strong constitutive promoter and the other under IPTG inducible promoter. We experienced troubles expressing the protein with strong promoters, it is possible that the protein can be toxic to the cell in high concentrations therefore we would suggest using weaker promoters.Biobrick is cloned between XbaI and SpeI sites, however it is cloned in reverse orientation due to XbaI and SpeI enzymes having the same overhangs after restriction digest. To use this biobrick future teams should use PCR apmlification rather than restriction digest to clone this biobrick. <html><a href="http://2016.igem.org/Team:Sheffield/project/science/furreporter">Read more about Mc.</a></html>
 <span class='h3bb'>Sequence and Features</span>
 <partinfo>BBa_K2016003 SequenceAndFeatures</partinfo>
 ===Functional Parameters===
 <partinfo>BBa_K2016003 parameters</partinfo>