Difference between revisions of "Part:BBa K2230021"

 
Line 3: Line 3:
 
<partinfo>BBa_K2230021 short</partinfo>
 
<partinfo>BBa_K2230021 short</partinfo>
  
−
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.  
+
''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.
 +
 
 +
[[File:Mingdaophil1026-2.png|550px|center]]
 +
 
 +
 
 +
 
 +
=== Cloning ===
 +
The ptsG gene was amplified from gDNA of ''Salmonella typhimurium'' using a primer with RBS sequence (B0034), and then cloned onto pSB1C3
 +
 
 +
[[File:Mingdaophil1026-3.jpeg|400px|center]]
 +
 
 +
 
 +
 
 +
=== 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.
 +
 
  
−
Cloning: The ptsG gene was amplified from gDNA of Salmonella typhimurium using a primer with RBS sequence (B0034), and then cloned onto pSB1C3
 
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 05:31, 26 October 2017


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.

Mingdaophil1026-2.png


Cloning

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

Mingdaophil1026-3.jpeg


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]