Part:BBa_K3282002

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 (Part:BBa_K3282005)and (Part:BBa_K3282007) expressed in pUC19 backbone. The composites have the same RBS (Part:BBa_J61109) and terminators (Part:BBa_B0012) with two different promoters T7 (Part:BBa_I719005) and Tac (Part: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]