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Definitions

• I-Dmo I when the 23S rRNA gene is transcribed into pre-23S rRNA, after which the intron is excised, it circularizes by ligation of the ends, and then the enzyme, I-Dmo I, is expressed by translation from nucleotide 59 through nucleotide 622 to nucleotide 21 (stop codon) thus having the amino acid sequence stated in SEQ ID No. 3.
• I-Dmo I has been proven a sequence-specific endonuclease in the following. However, as also shown in the following, the truncated peptide stated in SEQ ID No.
• Fig. 8 Autoradiograms of RNA sequencing gels demonstrating the circularity of the introns. Sequences were generated using the reverse transcriptase procedure as described in the legend to Fig. 6. The ligated junctions are indicated by arrows, and the bordering sequences corresponding to the rRNA template are given. No termination of reverse transcription was observed at the junctions. RNA was isolated as described in the legend to Fig. 5.
• Fig. 21 Cleavage of a DNA substrate from the intron- Pb. islandicum by I-Por I.
• A Gene map showing the location of a 23S rDNA fragment on a Cla I fragment (C-C) isolated from M13mp18. Arrows indicate the conserved sequence positions where the rRNA introns of Pyrobaculum organotrophum (Pb. o.) (intron 1) and Desulfurococcus mobilis (Dc. m. ) lie.
• the 23S rRNA intron of the archaeon Desulfuroccus mobilis circularizes after excision from pre-23S rRNA and forms a normal 5'-3'-phosphodiester bond at the ligation junction.
• This circular intron (622 nt) contains a putative ORF which crosses the ligation junction and covers most of the intron (Kjems and Garrett, 1985; 1988).
• the intron generates a complex secondary structure which has now been determined experimentally (Fig. 2).
• the other characterized archaeal introns from tRNA-encoding genes and other rRNA-encoding genes (Kjems and Garrett, 1991) are small ( ⁇ 110 nt).
• organotrophum (Huber et al., 1987) yielded a larger PCR product after an increased extension time compatible with the presence of one or more inserts (Fig. 3).
• the sequence of this DNA product was determined and aligned with 23S rRNA-encoding gene sequences from other hyperthermophiles. This revealed that it contained two additional sequences which could constitute introns (Fig. 4); both exhibited ORF's which, in contrast to that of Dc. mobilis 23S rRNA (Kjems and Garrett, 1985), do not cross the insert junctions (Fig. 4).
• a Southern blotting analysis was made to determine the number of 23 rRNA-encoding genes present in the organism.
• introns 1 and 2 in the latest secondary structural model of the domain IV-V region of the 23S rRNA from Pb. organotrophum are shown in Fig. 9 together with the positions of the other known archaeal rRNA introns.
• Many of the eukaryotic group I and group II introns found in pre- 23S-like rRNAs also occur within this functionally important region of the rRNA (reviewed by Garrett et al., 1991).
• Media LB media containing 100 ⁇ g ampicilin/liter.
• AGT AGG GAG TTT TAC GAA TCT GTC AGA GGC GCC ATT GAG CGG CTT CTA 294 Ser Arg Glu Phe Tyr Glu Ser Val Arg Gly Ala Ile Glu Arg Leu Leu

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