Difference between revisions of "Part:BBa K3282002"

Revision as of 10:59, 16 October 2019

C. metallidurans lead binding protein - pbrD

This part contains coding sequence of Cupriavidus metallidurans CH34 lead binding protein (pbrD, Gene ID: 3170401, Uniprot: Q58AJ8) codon-optimized for Escherichia coli Nissle 1917. Pb(II) resistance was early recognized to be located on pMOL30, one of the two plasmids found in Cupriavidus metallidurans CH34 [1]. The pbrD gene encodes a Pb(II)-binding protein which is essential for functional lead sequestration. When Pb(II) enters the cytoplasm, it can be bound by the pbrD protein, which may function as a chaperone for Pb(II) and cells lacking pbrD show a decreased accumulation of Pb(II) and this accumulation may protect against free exported Pb(II) [3]. The part was evaluated in the composite parts BBa_K3282005and BBa_K3282007 expressed in pUC19 backbone (part---). The composites have the same RBS BBa_J61109 and terminators BBa_B0012 with two different promoters T7 BBa_I719005 and Tac BBa_K3282006. The four composite parts were tested to determine their Lead accumulation capability.

1. Taghavi, S., Lesaulnier, C., Monchy, S., Wattiez, R., Mergeay, M., & Lelie, D. V. D. (2008). Lead(II) resistance in Cupriavidus metallidurans CH34: interplay between plasmid and chromosomally-located functions. Antonie Van Leeuwenhoek, 96(2), 171–182. doi: 10.1007/s10482-008-9289-0.

2.Hobman, J. L., Julian, D. J., & Brown, N. L. (2012). Cysteine coordination of Pb(II) is involved in the PbrR-dependent activation of the lead-resistance promoter, PpbrA, from Cupriavidus metallidurans CH34. BMC Microbiology, 12(1), 109. doi: 10.1186/1471-2180-12-109

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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 <partinfo>BBa_K3282002 short</partinfo>
-This part contains coding sequence of Cupriavidus metallidurans CH34  lead binding protein (pbrD, Gene ID: 3170401, Uniprot: Q58AJ8) codon-optimized for Escherichia coli Nissle 1917. Pb(II) resistance was early recognized to be located on pMOL30, one of the two plasmids found in Cupriavidus metallidurans CH34 (1). The pbrD gene encodes a Pb(II)-binding protein which is essential for functional lead sequestration. When Pb(II) enters the cytoplasm, it can be bound by the PbrD protein, which may function as a chaperone for Pb(II) and cells lacking PbrD show a decreased accumulation of Pb(II) and this accumulation may protect against free exported Pb(II)(2).
+This part contains coding sequence of ''Cupriavidus metallidurans'' CH34  lead binding protein (''pbrD'', Gene ID: 3170401, Uniprot: Q58AJ8) codon-optimized for Escherichia coli Nissle 1917. Pb(II) resistance was early recognized to be located on pMOL30, one of the two plasmids found in ''Cupriavidus metallidurans'' CH34 [1]. The ''pbrD'' gene encodes a Pb(II)-binding protein which is essential for functional lead sequestration. When Pb(II) enters the cytoplasm, it can be bound by the ''pbrD'' protein, which may function as a chaperone for Pb(II) and cells lacking ''pbrD'' show a decreased accumulation of Pb(II) and this accumulation may protect against free exported Pb(II) [3].
+The part was evaluated in the composite parts  [https://parts.igem.org/Part:BBa_K3282005 BBa_K3282005]and [https://parts.igem.org/Part:BBa_K3282007 BBa_K3282007] expressed in pUC19 backbone (part---). The composites have the same RBS [https://parts.igem.org/Part:BBa_J61109 BBa_J61109] and terminators [https://parts.igem.org/Part:BBa_B0012 BBa_B0012] with two different promoters T7 [https://parts.igem.org/Part:BBa_I719005 BBa_I719005] and Tac [https://parts.igem.org/Part:BBa_K3282006 BBa_K3282006]. The four composite parts were tested to determine their Lead accumulation capability.