RRC ID 33496
Author Krieg PA, Melton DA.
Title Functional messenger RNAs are produced by SP6 in vitro transcription of cloned cDNAs.
Journal Nucleic Acids Res
Abstract We describe a method for the synthesis of microgram quantities of eucaryotic messenger RNAs. Injection into the cytoplasm of frog oocytes and addition to wheat germ extracts show that these synthetic RNAs function efficiently as messenger RNAs. We confirm that a 5' cap on the mRNA is essential for translation in injected oocytes and show that most of the 3' flanking region, including the poly A tail, can be deleted without the abolition of protein synthesis. The method of mRNA synthesis involves in vitro transcription of cDNAs which have been cloned into SP6 vectors (described in the accompanying paper). This method enables one to produce large amounts of mRNA and consequently protein from any cDNA clone.
Volume 12(18)
Pages 7057-70
Published 1984-9-25
DOI 10.1093/nar/12.18.7057
PMID 6207484
PMC PMC320142
MeSH Animals Cloning, Molecular* DNA / metabolism* Female Globins / genetics Interferons / genetics Oocytes / metabolism Plants / metabolism Protein Biosynthesis RNA, Messenger / genetics* Salmonella Phages / genetics* Salmonella typhimurium / genetics* Transcription, Genetic* Triticum / metabolism Xenopus
IF 11.502
Times Cited 1336
Prokaryotes E. coli pSP64 pSP65