Home LiteratureArticle Details
PMID: 6294601 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Simple, efficient in vitro synthesis of capped RNA useful for direct expression of cloned eukaryotic genes.

Nucleic acids research ·Vol. 10 ·No. 20 ·1982-10-25 ·Pages 6353-62

Contreras R, Cheroutre H, Degrave W, Fiers W

Abstract

A simple and efficient method for direct in vitro synthesis of capped transcripts of cloned eukaryotic genes is described. As an example capped transcripts were made from a plasmid containing the human fibroblast interferon gene cloned under the control of a prokaryotic promoter. These transcripts were translated in vivo in Xenopus laevis oocytes and in vitro in reticulocyte and in wheat germ cell-free protein synthesizing systems.

MeSH Terms
Animals Cloning, Molecular DNA Restriction Enzymes Female Genes Oocytes/metabolism Plants/metabolism Plasmids Protein Biosynthesis RNA Caps/chemical synthesis Reticulocytes/metabolism Transcription, Genetic Triticum/metabolism Xenopus
Chemicals
RNA Caps DNA Restriction Enzymes
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Contreras R
Cheroutre H
Degrave W
Fiers W
References (21)
21 references, click to expand
  1. Studies on the kinetics of ribonucleic acid chain initiation and elongation.
    Biochemistry. 1970 Jun 9;9(12):2520-5 PMID: 4912487
  2. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  3. Sequence studies of satellite tobacco necrosis virus RNA. Isolation and characterization of a 5'-terminal trinucleotide.
    J Mol Biol. 1968 Nov 28;38(1):59-73 PMID: 5760635
  4. Determination of nucleotide sequences at promoter regions by the use of dinucleotides.
    Biochemistry. 1971 Dec 21;10(26):4970-5 PMID: 4944064
  5. Studies on polynucleotides. CXIX. Transcription of short double-stranded deoxyribonucleic acids of defined nucleotide sequences.
    J Biol Chem. 1972 Oct 10;247(19):6149-56 PMID: 4568604
  6. Studies on polynucleotides. CXX. On the transcription of a synthetic 29-unit long deoxyribopolynucleotide.
    J Biol Chem. 1972 Oct 10;247(19):6157-66 PMID: 4568605
  7. The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites.
    Proc Natl Acad Sci U S A. 1974 Apr;71(4):1342-6 PMID: 4598299
  8. Efficient translation of tobacco mosaic virus RNA and rabbit globin 9S RNA in a cell-free system from commercial wheat germ.
    Proc Natl Acad Sci U S A. 1973 Aug;70(8):2330-4 PMID: 4525168
  9. Methylation of newly synthesized viral messenger RNA by an enzyme in vaccinia virus.
    Proc Natl Acad Sci U S A. 1974 Aug;71(8):3014-8 PMID: 4606808
  10. Analysis of 32P-labeled bacteriophage MS2 RNA by a mini-fingerprinting procedure.
    Anal Biochem. 1976 May 7;72:433-46 PMID: 942063
  11. Proceedings: Controlled transcription of the synthetic gene for tyrosine transfer RNA.
    Arch Int Physiol Biochim. 1976 Feb;84(1):145-7 PMID: 60927
  12. Nucleotide sequence of the restriction fragment Hind F-Eco RI2 of SV40 DNA.
    Nucleic Acids Res. 1977 Apr;4(4):1001-14 PMID: 194225
  13. Complete nucleotide sequence of SV40 DNA.
    Nature. 1978 May 11;273(5658):113-20 PMID: 205802
  14. Analysis of RNA initiated in isolated mouse myeloma nuclei using purine nucleoside 5'[gamma-S]triphosphates as affinity probes.
    Cell. 1978 Oct;15(2):615-26 PMID: 719754
  15. How do eucaryotic ribosomes select initiation regions in messenger RNA?
    Cell. 1978 Dec;15(4):1109-23 PMID: 215319
  16. Efficient translation of prokaryotic mRNAs in a eukaryotic cell-free system requires addition of a cap structure.
    Nature. 1979 Jun 21;279(5715):692-6 PMID: 450118
  17. Isolation and structure of a human fibroblast interferon gene.
    Nature. 1980 Jun 19;285(5766):542-7 PMID: 6157094
  18. Expression of human fibroblast interferon gene in Escherichia coli.
    Nature. 1980 Sep 18;287(5779):193-7 PMID: 6159534
  19. A simple apparatus for injection of nanoliter quantities into Xenopus laevis oocytes.
    Anal Biochem. 1981 May 1;113(1):185-7 PMID: 7270884
  20. Plasmid vectors for high-efficiency expression controlled by the PL promoter of coliphage lambda.
    Gene. 1981 Oct;15(1):81-93 PMID: 6271633
  21. Systematic alteration of the nucleotide sequence preceding the translation initiation codon and the effects on bacterial expression of the cloned SV40 small-t antigen gene.
    Gene. 1982 Jan;17(1):55-63 PMID: 6281136
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/10.20.6353
Published
1982-10-25
Pages
6353-62
Language
English
Region
England
NLM ID
0411011
PMCID
PMC326924
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/10.20.6353 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2