DNA encoding m12Smith (residues 29-208) was ligated into the pFastbac vector (ThermoFisher Scientific) upstream of a 6x Histidine tag and used to generate recombinant baculovirus according to the manufacturer’s instructions. Specifically, the plasmid was transformed into DH10Bac E.coli cells in ...
Truncated, wild-type XylE (residues 6–480) gene was PCR amplified from the genomic DNA of theE. colistrain BL21(DE3). The gene was subcloned into pET-15b vector (EMD Millipore) and transformed intoE. colistrain BL21 C43(DE3). TheE. colicontaining the XylE expression plasmid was cultured...
the integrated suicide plasmid was forced to recombine out of the chromosome by growing on agar plates containing 10% sucrose. Mutants were selected based on their survival on plates containing 10% sucrose and loss of the chloramphenicol resistant marker of vector pDM4. The mutations were confirmed...