Part:BBa_M50051
IPTG Inducible Plasmid for the Expression of LsrR Repressor
This is a IPTG-inducible plasmid that encodes the expression of an LsrR repressor for the Lsr operon in E. coli. Although there are endogenous sources of LsrR, the plasmid allows for a controlled over-expression of protein through IPTG. This allows for the artificial control of intake of autoinducer 2(AI-2). The LsrR is a repressor for the lsr operon coding for the key components in the intake of AI-2(through membrane proteins), and its digestion in the cells. With regulation of this pathway, this should theoretically allow for control of the AI-2 quorum sensing, and in turn how cells sense each other. The promoter, the RBS, the his-tag, and the terminator were sourced through DNA 2.0. However, we have included parts from the Parts Registry in this project. The promoter is a T5 IPTG-inducible promoter that allows for the control of the expression of LsrR by varying the induction. The his-tag will allow us to test the expression through a Western blot.
Usage and Biology
LsrR is a repressor protein that inhibits the expression of the Lsr operon. By preventing the expression of the Lsr operon, LsrR reduces the intake of Autoinducer-2, an important quorum sensing molecule that is conserved throughout a wide variety of bacteria species.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal SpeI site found at 1134
- 12INCOMPATIBLE WITH RFC[12]Illegal SpeI site found at 1134
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal SpeI site found at 1134
- 25INCOMPATIBLE WITH RFC[25]Illegal SpeI site found at 1134
- 1000COMPATIBLE WITH RFC[1000]
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