Team:TU Darmstadt/Notebook/sec1/K1602017
K1602017 - D-Xylonic Acid Producing Operon (T7-B0034-xylB-B0034-xylC)
Figure 1 Gel of the CPCR of several clones. T7-B0034-xylB-B0034-xylC string has a length of approximately 2 kbp if you use VF2 and VR oligonucleotides.
The BBa_K1602016 part was combined with a T7-promotor for enabling protein expression.
B0034-xylB-B0034-xylC-pSB1C3 plamid was cut with XbaI and PstI and ligated into a T7-RFP-pSB1A2 plasmid which was cut with EcoRI and PstI. E. coli Top 10 cells were transformed with the ligation product. The resulting colonies were screened for positive clones by colony PCR using VF2 and VR oligonucleotides. Plasmid DNA was isolated from positive clones and verified by sanger sequencing. In addition to that cells were induced with IPTG for 12 hours at 28°C. The expressed protein was validated with SDS-PAGE.