FIG. 2.
Analysis of the as-48AB cotranscription. (A) Northern blot analysis. RNA was isolated from E. faecalis S-48[pMB2] and JH2-2 carrying the indicated pAM401-derived plasmid at the end of the exponential phase. The upper panel shows an autoradiogram of the blotting. The lines labeled 16S rRNA and 23S rRNA indicate the locations of these rRNA species, which are known to trap smaller RNA species (25). The lower panel shows the 23S rRNA and 16S rRNA loaded in the gel and stained with ethidium bromide. TA, RNA species containing the as-48A gene (see the legend of Fig. 5 for details). (B) Semiquantitative RT-PCR detection. RNA was isolated from E. faecalis JH2-2 carrying pM401-81 wild-type and mutant plasmids. The specific primers used and reaction conditions are described in Materials and Methods. The plasmid carried by the JH2-2 strain and the number of cycles of each sample are indicated.