Oligonucleotides used in this study.

aItalicized letters indicate restriction enzymes used in making constructs. Primer pairs used for construction of an in-frame pplA deletion mutant were pplA-SoeA-XbaI and pplA-SoeB to amplify the upstream flanking region and pplA-SoeC and pplA-SoeD-EcoRI to amplify the downstream flanking region, both pplA-SoeB and pplA-SoeC contain internal KpnI sites for insertion of the ermB gene (similar combination of primer pairs were used for construction of the respective pplA-G72 stop codon mutant and the eep deletion mutant, but eep-SoeA contained a PstI site and eep-SoeD contained a SacI site). oppA gene-disruption primers contained a KpnI site, pIMK2C’F a NcoI site, pIMK2C’R a XmaI site, pplAHis-F a KpnI site, and pplAHis-R an XmaI site.
bLetters in bold indicate the premature stop codon engineered at amino acid position G72 in the pplA coding sequence.
Oligonucleotides used in this study.