Figure 6.

Primer extension reactions on total RNA isolated from pneumococcal cells harbouring plasmid pMVT-PfcsK. Bacteria were grown in the absence or presence (1%) of fucose. As primers, different 5΄-radioactively labelled oligonucleotides were used: primer PE-tet (lanes 1 and 5); primer PE-mobM (lanes 2 and 6); primer PE-rnaII (lanes 3 and 7); a mix of the PE-tet, PE-mobM, PE-cop-rep and PE-rnaII primers (lanes 4 and 8). Primers were designed taking into account the transcription start site of the genes under study. Primer extension products were analysed by 8 M urea–8% polyacrylamide gel electrophoresis. Sequence ladders were used as DNA size markers (lanes A, C, G and T). They were prepared using M13 DNA and forward sequencing primer (Table 2). The sizes (in nucleotides) of the cDNA extension products are indicated on the right.