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Effect of template topology and temperature on in vitro transcription from the S. shibatae 16S rRNA promoter by using a reconstituted (a) or unfractionated (10 μg of Sulfolobus crude cell extract) system (b). Transcription of negatively supercoiled template (lanes 1 and 3) or linearized template (lanes 2 and 4) was as described in Materials and Methods. Transcription reactions were incubated at 75°C (lanes 1 and 2) or 48°C (lanes 3 and 4) and RNA detected by primer extension.
