Overproducing strain construction and purification of protein RM.Eco29kI. A - eco29kIR and M ORFs orientation on the natural plasmid pECO29 . On the plasmid Stop codon of eco29kIR gene overlaps Start codon of eco29kIM gene. B - A scheme of site-directed mutagenesis used for generation of the fusion protein RM.Eco29kI. Overlapped Stop and Start codons were substitutes for two Glycine codons. C - 10% PAGE electrophoresis with RM.Eco29kI induction (lane 2); final preparation of the enzyme (lane 3). Lane 1 - protein extract from non-induced cells, M - Dalton markers of 26, 34, 47, 86 and 120 kDA.
Mokrishcheva et al. BMC Evolutionary Biology 2011 11:35 doi:10.1186/1471-2148-11-35