1984
DOI: 10.1002/j.1460-2075.1984.tb02271.x
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A novel in vitro transcription-translation system: accurate and efficient synthesis of single proteins from cloned DNA sequences.

Abstract: A system is described which permits the efficient synthesis of single proteins in vitro. The essential element in this expression system is a strong promoter derived from coliphage T5 which produces, with high efficiency, specific RNAs in capped or uncapped form, depending upon the experimental conditions used. The transcription‐coupled capping of RNA allows the direct translation of the RNA in eukaryotic extracts from wheat germ as well as from HeLa cells. The synthesis of three different proteins is reported… Show more

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Cited by 251 publications
(156 citation statements)
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“…Radiolabeled precursor proteins were synthesized in reticulocyte lysate as described previously (Pelham and Jackson, 1976;Krieg and Melton, 1964;Stueber et al, 1964).…”
Section: Synthesis Of Radiolsbsled Proteins and Import In Vitromentioning
confidence: 99%
“…Radiolabeled precursor proteins were synthesized in reticulocyte lysate as described previously (Pelham and Jackson, 1976;Krieg and Melton, 1964;Stueber et al, 1964).…”
Section: Synthesis Of Radiolsbsled Proteins and Import In Vitromentioning
confidence: 99%
“…Though, in these cases transcripts were "capped" by the addition of 7-methylguanosine triphosphate to the transcription mixture (18) no proteins were synthesized in the rabbit reticulocyte lysate and only low amounts of proteins were produced in the wheat germ extract (data not shown). $30 lysates prepared from intact chloroplasts (1) of spinach and barley did not show any protein synthesis due to exogenously added DNA or transcripts derived from pDS6 (data not shown).…”
Section: Resultsmentioning
confidence: 99%
“…DNA-fragments containing either both cytochrome b-559 genes together or the individual psbE and psbF were inserted into the multiple cloning sites of the expression vectors pDS6 (18) and pGem3 (Promega Biotec) (Figure 1). Transcripts made from pDS6 and pGem3 were translated in an $30 lysate prepared from E. coli cells (14,22).…”
Section: Resultsmentioning
confidence: 99%
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