Difference between revisions of "Part:BBa K3282001"

Latest revision as of 16:36, 17 October 2019

C. metallidurans lead transport protein - pbrT

This part contains coding sequence of Cupriavidus metallidurans CH34 lead transport protein (pbrT, Gene ID: 3170370, Uniprot: Q58AJ4) codon-optimized for Escherichia coli Nissle 1917. pbrT is a lead(II) uptake protein containing a cytochrome domain and a transmembrane metal transport domain [1]. It belongs to the ILT (Iron Lead Transporter) family. This protein is a part of the lead (Pb) resistance operon in plasmid pMOL30 [2]. Expression of pbrT leads to uptake of lead into the cytoplasm to reduce interaction of free Pb(II) with side chains of membrane and periplasmic proteins, which would cause extensive cellular damage [3]. The part was evaluated in the composite parts BBa_K3282005 and BBa_K3282007 expressed in pUC19 backbone. The composites have the same RBS (BBa_J61109), terminator (BBa_B0012) with two different promoters T7 (BBa_I719005) and Tac (BBa_K3282006). The four composite parts were tested to determine their Lead uptake capability.

References

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, PbrA, from Cupriavidus metallidurans CH34. BMC Microbiology, 12(1), 109. doi: 10.1186/1471-2180-12-109

3. Borremans, B., Hobman, J. L., Provoost, A., Brown, N. L., & van Der Lelie, D. (2001). Cloning and functional analysis of the pbr lead resistance determinant of Ralstonia metallidurans CH34. Journal of bacteriology, 183(19), 5651–5658. doi:10.1128/JB.183.19.5651-5658.2001

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 643
Illegal NheI site found at 841
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 591
Illegal NgoMIV site found at 1054
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI site found at 1198
Illegal SapI.rc site found at 1428

@@ Line 4: / Line 4: @@
 This part contains coding sequence of ''Cupriavidus metallidurans'' CH34  lead transport protein (''pbrT'', Gene ID: 3170370, Uniprot: Q58AJ4) codon-optimized for ''Escherichia coli'' Nissle 1917. ''pbrT'' is a lead(II) uptake protein containing a cytochrome domain and a transmembrane metal transport domain [1]. It belongs to the ILT (Iron Lead Transporter) family. This protein is a part of the lead (Pb) resistance operon in plasmid pMOL30 [2]. Expression of ''pbrT'' leads to uptake of lead into the cytoplasm to reduce interaction of free Pb(II) with side chains of membrane and periplasmic proteins, which would cause extensive cellular damage [3].
-The part was evaluated in the composite parts BBa_K3282005 and BBa_K3282007 expressed in pUC19 backbone (part---). The composites have the same RBS (BBa_J61109), terminator (BBa_B0012) with two different promoters T7 (BBa_I719005) and Tac (BBa_K3282006). The four composite parts were tested to determine their Lead uptake capability.
+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. The composites have the same RBS ([https://parts.igem.org/Part:BBa_J61109 BBa_J61109]), terminator ([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 uptake capability.
 References