Figure 7
Accumulation of rps8*, an internal transcript within the rps8/spc operon covering part of the rps8 gene and the rpl5-rps8 intergenic spacer. (A) Accumulation of rps8* in the ethanol producer (Pr1 and Pr2) but not in the control strain (Co1 and Co2) after 14 and 18 days of cultivation in Crison photobioreactors. Total RNA was isolated from duplicate cultures, blotted and hybridized with a strand-specific RNA probe antisense to the rps8 sequence. For loading control, blots were hybridized with a probe against the 5S rRNA. (B) 5′RACE mapping of rps8* 5′ ends revealed an initiation of transcription 91 to 96 nt upstream of the rps8 start codon; plus sign (+) indicates treatment of RNA samples with tobacco acid pyrophosphatase; minus sign (−) indicates mock treatment. The bands excised for cloning and sequence analysis is indicated by the arrow. (C) Location of rps8* (arrow) within the rps8/spc operon.