Figure 2

Optimised linear templates for E. coli in vitro expression. Three kinds of linear PCR-products were generated to investigate the effect on protein expression rate and yield. (i) C-terminal wild type with C-terminal 6xHis-tag (ii) C-terminal mutant with C-terminal 6xHis-tag and inserted silent mutation at the N-terminus (iii) N-terminal wild type with N-terminal 6xHis-tag and no attachment sites (att-sites); For the C- and N-terminal wild type template the ORF is identical to the original ORF in the plasmid DNA.