Part:BBa_K5248053

pACYCDuet-1-KpsE-KpsT

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          Design: In podocarp research, the 2023 XJTU-iGEM team has used pgmA and galU gene overexpression to enable an increase in extracellular polysaccharide (EPS) production, based on which we additionally introduced overexpression of KpsE, KpsT, and FliC genes, which are capable of increasing the rate of podocarp polysaccharide translocation from intracellular to extracellular, thus optimizing our podocarp level of Defense.

Construct: The expression vector pACYCDuet-1-KpsE-KpsT was constructed, and the constructed plasmid and its empty vector were introduced into Nissle 1917 (DE3), respectively.    

 

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          Lane M: DNA Marker Lane 1: Plasmid digested by EcoRV and XhoI Lane 2: Uncut plasmid DNA Test: We carried out bacterial pod polysaccharide extraction on the above modified bacteria and measured the content of the extracted polysaccharide, using the empty vector-imported bacteria as the control, as shown in Fig. 1, our constructed KpsE-KpsT genetically engineered bacterium showed a significant increase in pod production compared with the empty vector-imported bacterium.    

 

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 1942
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 715
  Illegal XhoI site found at 1892
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1188
  Illegal SapI.rc site found at 1535