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Titolo:
Construction of promoter probe vectors for Synechocystis sp PCC 6803 usingthe light-emitting reporter systems Gfp and LuxAB
Autore:
Kunert, A; Hagemann, M; Erdmann, N;
Indirizzi:
Univ Rostock, FB Biol, D-18051 Rostock, Germany Univ Rostock Rostock Germany D-18051 , FB Biol, D-18051 Rostock, Germany
Titolo Testata:
JOURNAL OF MICROBIOLOGICAL METHODS
fascicolo: 3, volume: 41, anno: 2000,
pagine: 185 - 194
SICI:
0167-7012(200008)41:3<185:COPPVF>2.0.ZU;2-A
Fonte:
ISI
Lingua:
ENG
Soggetto:
GREEN FLUORESCENT PROTEIN; CIRCADIAN GENE-EXPRESSION; SP STRAIN PCC-6803; PHOTOSYSTEM-II; CYANOBACTERIUM; SYNECHOCOCCUS; LUCIFERASE; RHYTHMS; CLONING;
Keywords:
iron stress; GFP; LuxAB; promoter probe vectors; salt stress; Synechocystis;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Agriculture,Biology & Environmental Sciences
Life Sciences
Citazioni:
24
Recensione:
Indirizzi per estratti:
Indirizzo: Kunert, A Univ Rostock, FB Biol, Doeberaner Str 143, D-18051 Rostock, Germany Univ Rostock Doeberaner Str 143 Rostock Germany D-18051 Germany
Citazione:
A. Kunert et al., "Construction of promoter probe vectors for Synechocystis sp PCC 6803 usingthe light-emitting reporter systems Gfp and LuxAB", J MICROB M, 41(3), 2000, pp. 185-194

Abstract

Two promoter probe vectors were constructed for the cyanobacterium Synechocystis sp. strain PCC 6803 using reporter genes, which can be easily detected and quantified in vivo by the ability of their encoded proteins to emit light. The vectors allow the transcriptional fusion of promoter sequences with the gfp and luxAB genes, respectively, and their stable integration into a neutral site of the Synechocystis chromosome. Functionality df these vectors was demonstrated by cloning the promoter of the isiAB operon into both promoter probe vectors and analyzing the stress-dependent emission of light by the obtained reporter strains. As was found before for the isiAB operon, the P-isiAB reporter gene fusions were induced by iron starvation and high salt stress. Induction rates of mRNA of the wild type operon and the reporter gene fusions were found to be essentially the same, indicating that a promoter Fragment containing all necessary regulatory elements has been cloned. However, using the gfp gene a slow increase of protein and fluorescence was found, while the luxAB reporter gene constructs led to a rapid increase in luminescence. The same was found after retransfer of cells back into control media, in which the Gfp protein disappeared slowly, while the LuxAB-based luminescence decreased rapidly. These experiments show that both reporter genes can be used in Synechocystis: the luxAB system seems to be favourable regarding reaction time, while the gfp system has the advantage ofbeing independent from any substrate. (C) 2000 Elsevier Science B.V. All rights reserved.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 27/11/20 alle ore 01:52:03