Team:ETH Zurich/Composite Part
Composite Parts
Overview
All parts were codon-optimized for expression in E. coli Nissle with Geneious and modified to remove forbidden restriction sites.
| Part | Description | BioBrick |
|---|---|---|
| pConstRBSeng_TlpA | Best Composite Part: Constitutively expressed temperature-dependent transcriptional repressor of pTlpa with engineered RBS | BBa_K2500008 |
| pConst_RBS_LldP/ LldR_pConst_RBS_LuxR |
Expression cassette consisting of LuxR and LldP/LldR | BBa_K2500013 |
Design
BBa_K2500008: pConst_RBSeng_TlpA
Design notes: A RBS library was created using the Red Libs algorithm to find variants translating more TlpA RNA in order to create a thermoswitch tight enough to repress the premature induction of cell lysis by protein E.
Organism: Salmonella typhimurium
Based on: BBa_J23104 (promoter) and Piraner, Dan I., et al. Nature Chemical Biology 13.1 (2017): 75-80. (TlpA)
Source: FIXME RedLibs + gBlock
Registry: BBa_K2500008
BBa_K2500013: pConst1_RBS_LldP/LldR_pConst2_RBS_LuxR
Design notes:This expression cassette regulates the two inputs relayed to our AND gate, namely lactate and quorum sensing. LLdp is a L-lactate permease which transports lactate across the bacterial cell membrane. High lactate concentrations inhibit the LldR repressor proteins which under low-lactate conditions prevent access to the pLux quorum sensing promoter by LuxR-AHL complexes.
Higher bacterial cell densities result in a higher density of AHL communication molecules. AHL molecules diffuse across bacterial cell membranes, bind the transcription activator LuxR and activate the transcription of genes downstream of the pLux promoter, which is only accessible under high lactate levels as described above.
Organism: Escherichia coli, Vibrio fischeri
Based on: FIXME BBa_J23118 (pConst1), BBa_J23100 (pConst2), BBa_K1847007 (LldP/LldR) and BBa_C0062 (LuxR)
Source: FIXME
Registry: BBa_K2500013
