Part:BBa_K2016004
TdHr - iron-responsive, oxygen-responsive hemerythrin protein coding gene from Themiste dyscritum
TdHr is a protein expressed by Themiste dyscritum, a eukaryotic marine organism, in which this protein functions as an iron-binding transporter, much like our own haemoglobin. Td undergoes a slow process of oxygenation upon binding of iron and gains a violet/pink colour. This gene was codon-optimised for the use in E. coli and cloned into the pSB1C3 plasmid.
Usage and Biology
TdHr 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. Td has been shown to express relatively well using IPTG inducible system (Fig. 2). Expression has been established through SDS-PAGE gel, however due to time limitations, colour change in various iron concentrations has not been investigated. Read more about Td.
Figure 2: Expression of 3 hemerythrins: Dcr, Mc and Td in E. coli cells before and after induction with IPTG. For each protein 2 different biological repeats were first grown overnight and then induced with IPTG, L- ladder, U – uninduced, I – induced. For Dcr (16 kDa) and Td (13.5 kDa) we can see clear induction of expression after adding IPTG. Expression of Mc (14.7 kDa) is less clear and should be investigated further.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 338
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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