Functional messenger RNAs are produced by SP6 in vitro transcription of cloned cDNAs

Paul A Krieg, D. A. Melton

Research output: Contribution to journalArticle

1029 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)7057-7070
Number of pages14
JournalNucleic Acids Research
Volume12
Issue number18
DOIs
StatePublished - Sep 25 1984
Externally publishedYes

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Transcription
CDNA
Messenger RNA
Complementary DNA
Oocyte
Oocytes
Synthesis
3' Flanking Region
Protein Synthesis
Proteins
Wheat
Clone
Tail
Injection
RNA
Anura
Triticum
In Vitro Techniques
cDNA
Cytoplasm

ASJC Scopus subject areas

  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)
  • Genetics

Cite this

Functional messenger RNAs are produced by SP6 in vitro transcription of cloned cDNAs. / Krieg, Paul A; Melton, D. A.

In: Nucleic Acids Research, Vol. 12, No. 18, 25.09.1984, p. 7057-7070.

Research output: Contribution to journalArticle

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