eprintid: 8594
rev_number: 8
eprint_status: archive
userid: 1
dir: disk0/00/00/85/94
datestamp: 2009-03-02 11:01:21
lastmod: 2014-08-21 16:25:53
status_changed: 2012-08-14 15:28:11
type: article
metadata_visibility: show
creators_name: Stüber, Dietrich
creators_name: Ibrahimi, Ibrahim
creators_name: Cutler, Daniel
creators_name: Dobberstein, Bernhard
creators_name: Bujard, Herrmann
title: A novel in vitro transcription-translation system: accurate and efficient synthesis of single proteins from cloned DNA sequences
ispublished: pub
subjects: ddc-570
divisions: i-706000
keywords: transcription-coupled translation , T5 promoter , membrane translocation
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, including lysozyme, which is shown to be translocated across membranes when appropriate assay conditions are used. The simplicity of the experimental procedure, the high purity and specific activity of the [35S]methionine-labefled proteins produced offer a number of possibilities for the study of structure-function relationships of proteins.
abstract_translated_lang: eng
date: 1984
date_type: published
id_scheme: DOI
id_number: 10.11588/heidok.00008594
schriftenreihe_cluster_id: sr-2
schriftenreihe_order: 029
ppn_swb: 1373878355
own_urn: urn:nbn:de:bsz:16-opus-85945
language: eng
bibsort: STUBERDIETANOVELINVI1984
full_text_status: public
publication: The EMBO Journal
volume: 3
number: 13
pagerange: 3143-3148
citation:   Stüber, Dietrich ; Ibrahimi, Ibrahim ; Cutler, Daniel ; Dobberstein, Bernhard ; Bujard, Herrmann  (1984) A novel in vitro transcription-translation system: accurate and efficient synthesis of single proteins from cloned DNA sequences.  The EMBO Journal, 3 (13).  pp. 3143-3148.      
document_url: https://archiv.ub.uni-heidelberg.de/volltextserver/8594/1/Stueber_1984_EMBOJ.pdf