Inducible Expression and Splicing of Candida Group I Ribozyme in E.coli

【Abstract】 The Ca.LSU intron flanking a 129 bp exon upstream and a 100 bp exon downstream was inserted into the lacZ gene on pRS426 to transform E.coli . Northern blot analysis and RT\|PCR showed that splicing of Ca.LSU in E.coli is efficient upon inducible expression of the precursor RNA. In contrast, co\|transcriptional self\|splicing of the intron in vitro is much less active. Therefore, this E.coli splicing system can be used as a better model to investigate the effect of the ribozyme inhibitors on Ca.LSU splicing in living cell. We examined the effects of neomycin sulfate and pentamidine on Ca.LSU splicing in E.coli , and found that these drugs does\|dependently inhibit the intron splicing. However, heomycin is more potent than pentamidine in this action.更多还原