Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Plasmid cDNA-directed protein synthesis in a coupled eukaryotic in vitro transcription-translation system.
Craig D
,
Howell MT
,
Gibbs CL
,
Hunt T
,
Jackson RJ
.
???displayArticle.abstract???
A system is described in which transcription of cDNA clones by bacteriophage T7 RNA polymerase is coupled to translation in the micrococcal nuclease treated rabbit reticulocyte lysate in a single reaction of coupled transcription-translation. The monovalent and divalent cation requirements for translation are dominant for optimum expression in this coupled system, so that transcription is relatively inefficient. Nevertheless, the use of appropriate DNA concentrations leads to the synthesis of sufficient RNA to saturate the protein synthesis capacity of the system. The fidelity and efficiency of expression in this coupled system are high, and the degree of purification of the plasmid DNA is relatively uncritical. The system therefore offers very considerable advantages for rapid screening of 'mini-preparations' of cDNA plasmid constructs for retention of the correct reading frame and expression of the desired protein product.
Bergmann,
Translation of capped and uncapped vesicular stomatitis virus and reovirus mRNA'S. Sensitivity to m7GpppAm and ionic conditions.
1979, Pubmed
Bergmann,
Translation of capped and uncapped vesicular stomatitis virus and reovirus mRNA'S. Sensitivity to m7GpppAm and ionic conditions.
1979,
Pubmed
Borman,
Initiation of translation of human rhinovirus RNA: mapping the internal ribosome entry site.
1992,
Pubmed
,
Xenbase
Chu,
Translational recognition of the 5'-terminal 7-methylguanosine of globin messenger RNA as a function of ionic strength.
1978,
Pubmed
Dasso,
On the fidelity of mRNA translation in the nuclease-treated rabbit reticulocyte lysate system.
1989,
Pubmed
Dasso,
Efficient initiation of mammalian mRNA translation at a CUG codon.
1989,
Pubmed
Edery,
Functional characterization of eukaryotic mRNA cap binding protein complex: effects on translation of capped and naturally uncapped RNAs.
1984,
Pubmed
Elroy-Stein,
Cap-independent translation of mRNA conferred by encephalomyocarditis virus 5' sequence improves the performance of the vaccinia virus/bacteriophage T7 hybrid expression system.
1989,
Pubmed
Farrell,
Phosphorylation of initiation factor elF-2 and the control of reticulocyte protein synthesis.
1977,
Pubmed
Gehrke,
5'-Conformation of capped alfalfa mosaic virus ribonucleic acid 4 may reflect its independence of the cap structure or of cap-binding protein for efficient translation.
1983,
Pubmed
Hickey,
A relation between inhibition of protein synthesis and conformation of 5'-phosphorylated 7-methylguanosine derivatives.
1977,
Pubmed
Hunter,
The role of polyamines in cell-free protein synthesis in the wheat-germ system.
1977,
Pubmed
Jackson,
Potassium salts influence the fidelity of mRNA translation initiation in rabbit reticulocyte lysates: unique features of encephalomyocarditis virus RNA translation.
1991,
Pubmed
Jackson,
The novel mechanism of initiation of picornavirus RNA translation.
1990,
Pubmed
Jackson,
Preparation and use of nuclease-treated rabbit reticulocyte lysates for the translation of eukaryotic messenger RNA.
1983,
Pubmed
Jackson,
A detailed kinetic analysis of the in vitro synthesis and processing of encephalomyocarditis virus products.
1986,
Pubmed
Jang,
A segment of the 5' nontranslated region of encephalomyocarditis virus RNA directs internal entry of ribosomes during in vitro translation.
1988,
Pubmed
Kaminski,
Initiation of encephalomyocarditis virus RNA translation: the authentic initiation site is not selected by a scanning mechanism.
1990,
Pubmed
Kozak,
The scanning model for translation: an update.
1989,
Pubmed
Krieg,
In vitro RNA synthesis with SP6 RNA polymerase.
1987,
Pubmed
Mellits,
Removal of double-stranded contaminants from RNA transcripts: synthesis of adenovirus VA RNAI from a T7 vector.
1990,
Pubmed
Melton,
Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter.
1984,
Pubmed
Minshull,
Translation of cyclin mRNA is necessary for extracts of activated xenopus eggs to enter mitosis.
1989,
Pubmed
,
Xenbase
Monroy,
Purification of mRNA guanylyltransferase from vaccinia virions.
1978,
Pubmed
Monroy,
Characteristics of reactions catalyzed by purified guanylyltransferase from vaccinia virus.
1978,
Pubmed
Norman,
Isolation and properties of cDNA clones encoding SRF, a transcription factor that binds to the c-fos serum response element.
1988,
Pubmed
Palmenberg,
The nucleotide and deduced amino acid sequences of the encephalomyocarditis viral polyprotein coding region.
1984,
Pubmed
Pines,
Molecular cloning and characterization of the mRNA for cyclin from sea urchin eggs.
1987,
Pubmed
,
Xenbase
Rozen,
Bidirectional RNA helicase activity of eucaryotic translation initiation factors 4A and 4F.
1990,
Pubmed
Sonenberg,
Capped mRNAs with reduced secondary structure can function in extracts from poliovirus-infected cells.
1982,
Pubmed
Sonenberg,
Cap-binding proteins of eukaryotic messenger RNA: functions in initiation and control of translation.
1988,
Pubmed
van der Werf,
Synthesis of infectious poliovirus RNA by purified T7 RNA polymerase.
1986,
Pubmed
