Part:BBa_K2230021

RBS-ptsG/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.

Research

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 study, we demonstrated glucose absorption ability by overexpressing two systems in E. coli. Please go to our wiki page ([http://2017.igem.org/Team:Mingdao/Demonstrate Mingdao iGEM 2017]) for more information.

Cloning

The ptsG gene was amplified from gDNA of Salmonella typhimurium using a primer with RBS sequence (B0034), and then cloned onto pSB1C3

Reference

1. Glucose Galactose Malabsorption. American Journal of Physiology - Gastrointestinal and Liver Physiology 1998;275:G879-G882

2. Functional Properties and Genomics of Glucose Transporters. Curr Genomics. 2007;8(2): 113–128.

3. The SLC2 (GLUT) Family of Membrane Transporters. Mol Aspects Med. 2013;34(0): 121–138.

4. Glucose and Glycolysis Are Required for the Successful Infection of Macrophages and Mice by Salmonella enterica Serovar Typhimurium. Infect Immun. 2009;77(7): 3117–3126.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 801
  Illegal NgoMIV site found at 1224
  Illegal AgeI site found at 508
  
• 1000
  COMPATIBLE WITH RFC[1000]