Part:BBa_K1462410:Design

pTEF2+GroES+tADH1+pTEF2+GroEL+tADH1+pGAL1+PRK+GBD-ligand+tADH1+pTDH3+RuBisCo+SH3-ligand+tADH1+pTDH3+

Assembly Compatibility:

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

  Illegal BglII site found at 4138
  Illegal BglII site found at 8442
  Illegal BglII site found at 9468
  Illegal BamHI site found at 10286
  Illegal XhoI site found at 10267
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 3639
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

This part consists of promoter TEF2 + chaperon GroES + terminator ADH1 + promoter TEF2 + chaperon GroEL + terminator ADH1 + promoter GAL1+ PRK + the ligand of the scaffold GBD + terminator ADH1 + promoter TDH3 + RuBisCo + the ligand of the scaffold SH3 + terminator ADH1 + promoter TDH3 + CA + the ligand of the scaffold PDZ+ terminator ADH1

Figure 1. Overview of this part This is a contrast group without CA and the ligand of scaffold PDZ.</a>

Figure 2.The pre-experiment of fermentationTo test out whether CA can make a difference or not, we had two constrast recombinant groups, one(group Ⅰ) consisting of PRK-RuBisCo(in YEplac181) and GroES-GroEL(in YEp352), the other（group Ⅱ） consisting of PRK-RuBisCo-CA(in YEplac181) and GroES-GroEL(in YEp352) transfered into competent yeast cells. The colonies were first cultured in YPD media and then cultured in media containing galactose as carbon source, during which the yeast underwent a fermentation process and samples were taken out at a certain time for analysis. As shown below, at the beginning of the fermentation process, ethanol concentrations of contrast groups are higher than that of wild type cells（group 0）. With time going by, for group 0 and group Ⅰ ethanol concentrations increased first and then decreased. For group Ⅱ , on the contrary, ethanol concentration decreased first and then increased.

Source

Kits and a synthesis company.