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

Determination of the in vivo presence of pre-tRNAs and circular introns. (A) RT-PCR products using H. volcanii total RNA and various primers are separated on a native 6% polyacrylamide gel. The primers (Fig. 1) used, expected sizes of the products, and the template regions corresponding to these products (thick lines in the schematic diagrams) are indicated below the lanes. Lane 4 contains a 25-bp DNA ladder. (B) The 69-bp RT-PCR product in A, lane 5, is sequenced using primer L. The arrow denotes the position of the splice junction (Fig. 1F, asterisk) in the sequence of the circularized intron derived from elongator pre-tRNA^Met.