[Cleavage activity of ribozymes on a target RNA of tobacco mosaic virus in E. coli].
A target cDNA fragment from TMV RNA was inserted to a reporter gene CAT immediately to the 3' end of the translation initiation codon ATG resulting in the formation of a chimeric CAT gene in an in vivo transcription and expression vector. The in vivo activities of various ribozymes on the target sequence were observed by determining the changes of the CAT activities of the chimeric CAT gene expressed in E. coli. The CAT activity was reduced by up to 30% when a specific ribozyme RZ1, RZ1A or RZ1B was transcribed, no change in CAT activity was observed when a non-specific ribozyme RZ3 was expressed. The protein electrophoresis and primer extension experiments indicated that this reduction in CAT activity was due to the specific cleavage of the ribozymes to the chimeric CAT mRNA at the target region hence the decrease in CAT protein synthesis.
Zhang
Q Q
QQ
National Laboratory of Plant Molecular Genetics, Shanghai Institute of Plant Physiology, Chinese Academy of Sciences, Shanghai 200032.
Song
R T
RT
Xu
Z K
ZK
D019677:Artificial Gene Fusion; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis* / Q000235:genetics; D004926:Escherichia coli / Q000378:metabolism*; D016337:RNA, Catalytic / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D014027:Tobacco Mosaic Virus / Q000235:genetics*
D016337:RNA, Catalytic; D012333:RNA, Messenger; D015500:Chloramphenicol O-Acetyltransferase