Figure 5.

RT-PCR amplification of spliced B4-B5 exons. The first lane contains a DNA ladder of markers (Stratagene). The second and third lanes, respectively, represent RNA isolated from the yeast null strain (QBY320) that contained plasmids expressing wild-type LeuRS from yeast and M. tuberculosis. The unspliced and spliced B4-B5 exon products are indicated by respective bands at about 1.5 kbp and 250 bp. The fourth and fifth lanes, respectively, show that the intron remains unspliced when genes containing either the W286C mutant or G288SΔC5 deletion of M. tuberculosis LeuRS are used in attempts to complement the yeast null strain.