Part:BBa_K5115000
RcnR_C35L
Introduction
The transporter RcnA, already existing on the membrane of E. coli, has been implicated in Ni(II) detoxification in through efflux, which is the opposite of our objective. Luckily, overexpression of RcnR can inhibit the induction of rcnA by metal cations, helping us to retain the nickel ion inside our bacteria[1].The RcnR can regulate the expressing of RcnA by binding to its DNA. Typically, the DNA affinity of the proteins is altered in response to binding the regulated metal ion. In the case of RcnR, the protein responds to nickel ions by decreasing the affinity for DNA[2]. Introducing RcnR_C35L into our design, we hope that we can counteract the spontaneous nickel efflux of E. coli to the greatest extent.
Usage and Biology
In our part, we changed the only Cys residue, which is called Cys35 to be precise, to Leu. RcnR binds Ni(II) in six-coordinate sites with pseudo-octahedral geometry at subunit interface, Cys35 being one of the binding sites. The change of it can stop RcnR from decreasing its DNA affinity, stopping RcnA's responding to high nickel concentration. This change can eventually make sure that the already-absorbed nickel will not run away from our bacteria.[3] In our design, RcnR_C35L mainly works with BBa_K5115050(MTA, Metallothionein)and BBa_K1151001(hpn). They can raise the absorptivity of nickel ion as well as reduce the harm nickel brings to our engineering bacteria.
Sequence and Features
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 228
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
References
- ↑ Koch, D., Nies, D. H., & Grass, G. (2007). The RcnRA (YohLM) system of E. coli: A connection between nickel, cobalt and iron homeostasis. BioMetals, 20(5), 759–771.
- ↑ Huang, H.-T., & Maroney, M. J. (2019). Ni(II) Sensing by RcnR Does Not Require an FrmR-Like Intersubunit Linkage. Inorganic Chemistry, 58(20), 13639–13653.
- ↑ Huang, H.-T., & Maroney, M. J. (2019). Ni(II) Sensing by RcnR Does Not Require an FrmR-Like Intersubunit Linkage. Inorganic Chemistry, 58(20), 13639–13653.
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