Difference between revisions of "Part:BBa K1962004"
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===Usage and Biology=== | ===Usage and Biology=== | ||
| − | Our idea was to generate a device that would express Colicin E3 in the presence of low pH and bile salts | + | Our idea was to generate a device that would express Colicin E3 in the presence of low pH and bile salts. In order to protect the host cells from the toxic activity of Colicin E3 we began by cloning the E3-Immunity protein downstream of a pH sensitive promoter Pasr (BBa_K1231000) and a bile salt sensitive promoter PacrRA (BBa_K1231001) to generate the composite parts (<partinfo>BBa_K1962016</partinfo>) and (<partinfo>BBa_K1962012</partinfo>), respectively. Unfortunately we were unable to clone Colicin E3 downstream of the E3-Immunity proteins. |
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Revision as of 23:36, 22 October 2016
Immunity Protein Im-E3
This biobrick encodes for the Immunity protein specific for Colicin E3. This Immunity Protein inhibits the 16S RNA hydrolyzing activity of Colicin E3 by binding with high affinity to the C-terminal catalytic domain of E3. The sequence was obtained from Uniprot and then codon optimised for E. coli K-12 and synthesised by IDT as a gBlock gene fragment.
Usage and Biology
Our idea was to generate a device that would express Colicin E3 in the presence of low pH and bile salts. In order to protect the host cells from the toxic activity of Colicin E3 we began by cloning the E3-Immunity protein downstream of a pH sensitive promoter Pasr (BBa_K1231000) and a bile salt sensitive promoter PacrRA (BBa_K1231001) to generate the composite parts (BBa_K1962016) and (BBa_K1962012), respectively. Unfortunately we were unable to clone Colicin E3 downstream of the E3-Immunity proteins.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]