The Ca. LSU intron flanking a 129 bp cxon upstream and a fOO bp exondownstream was inserted into the lacZ gene on pRS426 to transform E. coli. Northern blot analysisand RT-PCR showed that splicing of Ca. LSU in E. col...The Ca. LSU intron flanking a 129 bp cxon upstream and a fOO bp exondownstream was inserted into the lacZ gene on pRS426 to transform E. coli. Northern blot analysisand RT-PCR showed that splicing of Ca. LSU in E. coli is efficient upon inducible expression of theprecursor RNA, In contrast, co-lranscriptional self-splicing of the intron in vitro is much lessactive. Therefore, this E. coli splicing system can be used as a better model to investigate theeffect of the ribozyme inhibitors on Ca. LSUsplicing in living cell. We examined (he effects ofneomycin sulfatc and pentamidine on Ca. LSU splicing in E. coli, and found that these drugsdoes-dependently inhibit the intron splicing. However, heomycin is more potent than pentamidine inthis action.展开更多
基金Supported by the National Natural Science Foundation of China (30170213) and the Foundation of Wuhan University(0000028)
文摘The Ca. LSU intron flanking a 129 bp cxon upstream and a fOO bp exondownstream was inserted into the lacZ gene on pRS426 to transform E. coli. Northern blot analysisand RT-PCR showed that splicing of Ca. LSU in E. coli is efficient upon inducible expression of theprecursor RNA, In contrast, co-lranscriptional self-splicing of the intron in vitro is much lessactive. Therefore, this E. coli splicing system can be used as a better model to investigate theeffect of the ribozyme inhibitors on Ca. LSUsplicing in living cell. We examined (he effects ofneomycin sulfatc and pentamidine on Ca. LSU splicing in E. coli, and found that these drugsdoes-dependently inhibit the intron splicing. However, heomycin is more potent than pentamidine inthis action.
