Designed by: BingZhao Zhuo, Yue Yang Group: iGEM16_NWPU (2016-10-11)
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.
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
- 10COMPATIBLE WITH RFC
- 12COMPATIBLE WITH RFC
- 21COMPATIBLE WITH RFC
- 23COMPATIBLE WITH RFC
- 25INCOMPATIBLE WITH RFCIllegal NgoMIV site found at 759
Illegal NgoMIV site found at 1221
Illegal AgeI site found at 223
- 1000INCOMPATIBLE WITH RFCIllegal SapI site found at 130