FIG. 5.

STM 1788 expression in wild-type and hya mutant cells. RNAs were extracted from wild-type and hya mutant cell cultures grown aerobically overnight in LB broth, using an Aurum Total RNA Mini kit (Bio-Rad). RNAs were digested with RQ1 DNase (Promega) to remove contaminating genomic DNA. These were used as templates to generate cDNAs by use of an iScript Select cDNA synthesis kit (Bio-Rad) and primers homologous to the gene directly downstream of the hya mutation (STM 1788). PCR was performed with the same primers used to generate cDNA. Lane 1, Promega 1-kb DNA ladder; lane 2, positive control (PCR product from wild-type genomic DNA); lanes 3 and 4, PCR products from wild-type and hya cDNAs, respectively; lanes 5 and 6, negative controls (PCR products from wild-type and hya cDNA reactions, respectively, where reverse transcriptase was not added to the mix).