Expression profiles of Tk-deaD at various growth temperatures. (A) Comparison of mRNA levels. RT-PCR reactions were performed with total RNAs from
LY450139 in logarithmic and stationary phases cultivated at 60 °C, 85 °C, and 93 °C. The levels of the Tk-deaD transcripts were evaluated as the signal intensities of fragments amplified with the respective gene-specific primers. As a control, the levels of 16S rRNA were examined. (B) Comparison of protein levels. Immunoblot analysis with anti-Tk-DeaD was performed using cytoplasmic extracts obtained from cells by the methods described in Materials and methods. (C) Mapping of the transcription start site for Tk-deaD. Primer extension reactions were performed with total RNAs isolated from T. kodakaraensis cultivated at 60 °C, 85 °C, and 93 °C. The extended reverse transcripts were analyzed with sequencing ladders generated using the same primer and template (lanes A, T, G and C). The transcription start site is represented by a triangle. The 5′-UTR is shaded. The TATA element is broken underlined. The initiation
codon is boxed. 16S rRNA binding region is underlined. The primer binding region is indicated by a broken arrow.
