Online citations, reference lists, and bibliographies.
← Back to Search

Preparative In Vitro MRNA Synthesis Using SP6 And T7 RNA Polymerases.

V. Gurevich, I. Pokrovskaya, T. Obukhova, S. A. Zozulya
Published 1991 · Chemistry, Medicine

Cite This
Download PDF
Analyze on Scholarcy
A method for preparative in vitro mRNA synthesis is presented. The method makes it possible to generate several hundred RNA copies per molecule of linearized DNA template. The protocol is applicable to the synthesis of RNAs of differing lengths (from 200 to 3000 bases), and both SP6 and T7 RNA polymerases can be used. The transcription can be scaled up to at least 6-12 ml, yielding 4.5-18 mg of homogeneous biologically active mRNA.
This paper references
Functional expression in vitro of bovine visual rhodopsin.
S. A. Zozulya (1990)
Bacteriophage SP6-specific RNA polymerase. I. Isolation and characterization of the enzyme.
E. T. Butler (1982)
Functional messenger RNAs are produced by SP6 in vitro transcription of cloned cDNAs.
P. Krieg (1984)
Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter.
D. Melton (1984)
Reticulocyte lysates synthesize an active alpha subunit of the stimulatory G protein Gs.
J. Olate (1988)
Purification of mouse immunoglobulin heavy-chain messenger RNAs from total myeloma tumor RNA.
C. Auffray (1980)
Ribonuclease inhibitor from human placenta: rapid purification and assay.
P. Blackburn (1979)
A continuous cell-free translation system capable of producing polypeptides in high yield.
A. Spirin (1988)
Cotranslational amino-terminal processing of cytosolic proteins. Cell-free expression of site-directed mutants of human hemoglobin.
J. P. Boissel (1988)
Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4
U. Laemmli (1970)
Regulation of LacZ mRNA translatability in a cell‐free system at heat shock by the last four sense codons
O. N. Denisenko (1989)
Molecular Cloning: A Laboratory Manual
J. Sambrook (1983)
Cell-free-synthesized interleukin-6 (BSF-2/IFN-beta 2) exhibits hepatocyte-stimulating activity.
T. Geiger (1988)

This paper is referenced by
Protein engineering ofDe novo protein with predesigned structure and activity
D. Dolgikh (1996)
Continuous-flow cell-free translation, transcription-translation, and replication-translation systems.
L. Ryabova (1998)
Polymorphism in the 3′-untranslated region of TNFα mRNA impairs binding of the post-transcriptional regulatory protein HuR to TNFα mRNA
S. Marco (2001)
Biophysical investigations of boranophosphate siRNA for use in RNA interference against human disease
Laura Moussa (2009)
BC1 RNA, the transcript from a master gene for ID element amplification, is able to prime its own reverse transcription.
M. Shen (1997)
Evidence for multiple mechanisms for membrane binding and integration via carboxyl-terminal insertion sequences.
P. K. Kim (1997)
A homogenous fluorescence assay of micro RNA maturation
Brian P. Davies (2008)
Cell‐free translation in reversed micelles
S. Nametkin (1992)
Kinetics of translation of γB crystallin and its circularly permutated variant in an in vitro cell‐free system: possible relations to codon distribution and protein folding
A. Komar (1995)
Specific RNA self-cleavage in coconut cadang cadang viroid: potential for a role in rolling circle replication.
Y. Liu (1998)
Structure determination and dynamics of protein-RNA complexes by NMR spectroscopy.
C. Dominguez (2011)
Paul N. Hengen (1996)
T7 RNA polymerase as a self-replicating label for antigen quantification.
Bakhos A. Tannous (2002)
Structural Requirements for the Binding of tRNA3Lys to Reverse Transcriptase of the Human Immunodeficiency Virus Type 1 (*)
B. B. Essink (1995)
Synthesis and maturation of green fluorescent protein in a cell-free translation system
V. Kolb (2004)
Modern methods for probing RNA structure.
J. Kjems (1998)
In vitro selective RNA synthesis with L-A virus nanoparticles.
T. Ebihara (1999)
Protein Engineering of Albebetin, A de Novo Protein with a Predesigned Three-Dimensional Structure
D. A. Dolgikh (1996)
Functional antibody production using cell-free translation: Effects of protein disulfide isomerase and chaperones
L. Ryabova (1997)
Is protein disulfide isomerase a redox‐dependent molecular chaperone?
Richard A Lumb (2002)
Interplay between Drosophila Bloom's syndrome helicase and Ku autoantigen during nonhomologous end joining repair of P element-induced DNA breaks
B. Min (2004)
Use of bacteriophage RNA polymerase in RNA synthesis.
V. Gurevich (1996)
A Site-specific, Membrane-dependent Cleavage Event Defines the Membrane Binding Domain of FtsY*
J. Millman (1999)
Cell-Free Synthesis Optimization of Barley Vacuolar Na/H-Antiporter, a Highly Hydrophobic Protein
A. Nizkorodova (2010)
Electrogenic amino acid exchange via the rBAT transporter
M. Coady (1994)
The 3′‐terminal untranslated region of alfalfa mosaic virus RNA 4 facilitates the RNA entry into translation in a cell‐free system
L. Ryabova (1993)
Proteins and azobenzenes
A. Reiter (2013)
The ribosome-in-pieces: binding of elongation factor EF-G to oligoribonucleotides that mimic the sarcin/ricin and thiostrepton domains of 23S ribosomal RNA.
A. Munishkin (1997)
Towards the Development of Synergistic Inhibitors that Exploit the Replication Strategy of HIV-1
L. K. Pattenden (2005)
rBAT is an Amino Acid Exchanger with Variable Stoichiometry
M. Coady (1996)
Novel hybridization assay configurations based on in vitro expression of DNA reporter molecules.
E. Laios (1998)
Identification of a Novel Function of the AlphavirusCapping Apparatus
L. Vasiljeva (2000)
See more
Semantic Scholar Logo Some data provided by SemanticScholar