... was amplified from E. coli K12 genomic DNA, includ-ing a C-terminal HA tag, using primers 5¢-GCGCGCGAATTCATGAGCATTATTAAAGAATTTCG-3¢ (forward)and 5¢-CGCGCGGGATCCTTAAGCATAATCAGGAACATCATAAGGATAACCACCAGGAGAGCGGTTATTCTGCTCTTTC-3¢ ... were 5¢-AGCAGAATAACTGCTCTCCTGGTG-3¢ (forward) and 5¢-CACCAGGAGAGCAGTTATTCTGCT-3¢ (reverse), and those for MscLF54C were 5¢-GGGATCGATTGCAAACAGTTTGC-3¢(forward) and 5¢-GCAAACTGTTTGCAATCGATCCC-3¢(reverse). ... Amsterdam, The Netherlands2 Bacterial Membrane Proteomics Laboratory, Instituto de Tecnologia Quı´mica e Biolo´gica, Avenida da Repu´blica, Estac a o Agrono´micaNacional, Oeiras, Portugal3...