Part:BBa_K2155013

BTT(BFD-TalB-TalB)

Usage and Biology

In order to simplify our experiment procedure in vivo, we fused BFD and TalB in one plasmid. And in consideration of TalB’s low catalytic efficiency compared with BFD, we inserted two copy of TalB’s sequence in this new plasmid(Figure1) so that intermediates produced by BFD could be transferred to D-xylulose in time. We cloned this part into the plasmid vector pET28a+ for expression. We verify this new part could be normally expressed in E.coli through SDS-PAGE. Following the expression test, the product are analyzed by HPLC(Figure2).

Figure1. Two copy of TalB’s sequence are inserted behind BFD’s sequence.

600px

Figure2. HPLC analysis of samples from in vivo experiment of BTT(BFD-TalB-TalB).

The HPLC results showed: I. The accumulation of two intermediate products obviously increased. II. the accumulation of D-xylulose was little.

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 759
  Illegal NgoMIV site found at 1221
  Illegal AgeI site found at 223
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 130