Two reactions of Haloferax volcanii RNA splicing enzymes: Joining of exons and circularization of introns

Ligase reactions using purified substrates. Gel-eluted endonuclease products of [α-³²P]CTP labeled pre-tRNA^Trp are used in ligase reactions. The products are separated by denaturing 6% PAGE. Lane 1, Pre-tRNA; lane 2, standard endonuclease reaction; lane 3, standard ligase reaction; lanes 4 and 6, gel-eluted linear introns and exons, respectively; lanes 5 and 7, ligase reactions using gel-eluted linear introns and exons, respectively. (P) Precursor; (LI) linear intron; (E) exons; (LE) ligated exons; (CI) circularized intron; (XC) xylene cyanol; (BB) bromophenol blue.