Functional messenger RNAs are produced by SP6 in vitro transcription of cloned cDNAs (original) (raw)
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Department of Biochemistry and Molecular Biology, Harvard University
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Department of Biochemistry and Molecular Biology, Harvard University
7 Divinity Avenue, Cambridge, MA 02138, USA
Search for other works by this author on:
Revision received:
07 September 1984
Accepted:
07 September 1984
Published:
25 September 1984
PDF Cite
P.A. Krieg, D.A. Melton, Functional messenger RNAs are produced by SP6 in vitro transcription of cloned cDNAs, Nucleic Acids Research, Volume 12, Issue 18, 25 September 1984, Pages 7057–7070, https://doi.org/10.1093/nar/12.18.7057
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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 invitro 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.
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