The resulting

purified cDNA was split to two reaction tubes and a homopolymeric A or G-tail was added to the 3′ end using recombinant terminal deoxynucleotidyl transferase and dATP or dGTP. A PCR product was amplified from each tailed cDNA using the appropriate anchor primer (oligo dT-AP or oligo dC-AP) and the nested gene specific primer BBB04 5′ RACE R2 2. The PCR products were subjected to electrophoresis and the bands were gel extracted using the QIAquick PCR Purification Kit (Qiagen). The sequence of the purified PCR products was determined using the BBB04 5′ RACE R2 primer and aligned to the upstream sequence of chbC to determine the transcriptional start site. Promoter analysis was carried out by visual EPZ-6438 chemical structure inspection and comparison selleck of the region upstream of the chbC Salubrinal manufacturer transcriptional start site with previously described RpoD, RpoS and RpoN-dependent promoter sequences in B. burgdorferi. Acknowledgements We thank P. Rosa, M. Norgard and J. Radolf for providing strains and plasmids. This research is based in part upon work conducted using the Rhode Island Genomics and

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