Difference between revisions of "Part:BBa K2230022"

Line 21: Line 21:
 
=== Cloning ===
 
=== Cloning ===
 
The STM1128 gene was amplified from gDNA of Salmonella typhimurium and cloned onto pSB1C3. This part has been sequenced.  
 
The STM1128 gene was amplified from gDNA of Salmonella typhimurium and cloned onto pSB1C3. This part has been sequenced.  
 +
 +
[[File:Mingdaophil1026-3.jpeg|600px]]
  
  

Revision as of 04:03, 26 October 2017


STM1128/pSB1C3


Mingdaophil1026-1.jpeg

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.


Research

Based on our research, the glucose transporter of Salmonella typhimurium has lowest Km, meaning the highest affinity of glucose bound to the transporter

Mingdaophil1026-2.png


In order to express the genes, we chose promoter CP29 that is a strong constitutive promoter working well in both E. coli and Lactobacillus spp. The biobrick part, CP29-RBS-aeBlue (BBa_K1033280), was used and assembled with the transporter genes.


Cloning

The STM1128 gene was amplified from gDNA of Salmonella typhimurium and cloned onto pSB1C3. This part has been sequenced.

Mingdaophil1026-3.jpeg


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 AgeI site found at 1255
  • 1000
    COMPATIBLE WITH RFC[1000]