Part:BBa_K2230027
CP29-RBS-aeBlue-RBS-crr-RBS-ptsG-TT/pSB1C3
Salmonella typhimurium LT2 has two glucose-specific transporter systems, PTS system and sodium/glucose cotransporter. PTS system contains two subunits IIA encoded by crr and IIBC by ptsG which are assembled to a high-affinity active transporter. The other is a Na+/glucose cotransporter encoded by STM1128 that contributes to facilitated transport with lower glucose affinity. Based on our research, the glucose transporter of Salmonella has a lower Km compared to human small intestine, Staphylococcus and E. coli, indicating a higher efficiency for glucose uptake. In our project, we created the glucose transporter device and genetically engineer microbes with these two systems.
In order to express the genes in E. coli for demonstration and in probiotics for proof-of-concept in a real world. We chose promoter CP29 that is a strong constitutive promoter working well in both E. coli and Lactobacillus spp1. The biobrick part, CP29-RBS-aeBlue (BBa_K1033280) was used and to be assembled with the transporter genes.
Cloning: Promoter CP29-RBS-aeBlue (BBa_K1033280) was assembled with RBS-crr-RBS-ptsG-TT/pSB1C3 (BBa_K2230026)
Function: An active high-affinity glucose transporter system. Promoter CP29 is a constitutive and strong promoter which works well in both E. coli and Lactobacillus spp. The blue protein encoded by aeBlue gene acts as indicator for the gene expression in the transgenic strain. This device can help E. coli and Lactobacillus spp. efficiently take up and retrieve glucose in the environment.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 2129
Illegal NgoMIV site found at 2552
Illegal AgeI site found at 976
Illegal AgeI site found at 1836 - 1000COMPATIBLE WITH RFC[1000]
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