摘要
Objective: To confirm the cleavage activit y of ribozyme Rz199 designed to oncogene Ki--ras messengerRNA in vitro, and survey the possibility of application of Rz199 in vivo. Methods: The plasmid for transcription invitro of ribozyme Rz199 and Ki ras exon 1 were constructed by DNA recombinant technique. Ribozyme Rz199 andKi--ras exon 1 mRNA were obtained by transcription in vtiro with T7 RNA polymerase. The cleavage reaction wasdone by mixing ribozyme Rz199 and its target RNA in a reactive buffer containing Mg2+. Results: The transcriptof 254 nucleotide (nt) of Ki--ras exon 1 was cleaved into 2 fragments of 90 and 164 nt by ribozyme Rz199.Conclusion: Ribozyme Rz199 can cleave Ki--ras mRNA in a site--specific manner in vitro. though its function in vivoremained to be studied further.
Objective: To confirm the cleavage activit y of ribozyme Rz199 designed to oncogene Ki--ras messengerRNA in vitro, and survey the possibility of application of Rz199 in vivo. Methods: The plasmid for transcription invitro of ribozyme Rz199 and Ki ras exon 1 were constructed by DNA recombinant technique. Ribozyme Rz199 andKi--ras exon 1 mRNA were obtained by transcription in vtiro with T7 RNA polymerase. The cleavage reaction wasdone by mixing ribozyme Rz199 and its target RNA in a reactive buffer containing Mg2+. Results: The transcriptof 254 nucleotide (nt) of Ki--ras exon 1 was cleaved into 2 fragments of 90 and 164 nt by ribozyme Rz199.Conclusion: Ribozyme Rz199 can cleave Ki--ras mRNA in a site--specific manner in vitro. though its function in vivoremained to be studied further.
