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Titolo:
Analysis of temporal and spatial expression of the CcaR regulatory elementin the cephamycin C biosynthetic pathway using green fluorescent protein
Autore:
Kyung, YS; Hu, WS; Sherman, DH;
Indirizzi:
Univ Minnesota, Dept Microbiol, Minneapolis, MN 55455 USA Univ Minnesota Minneapolis MN USA 55455 robiol, Minneapolis, MN 55455 USA Univ Minnesota, Biol Proc Technol Inst, Minneapolis, MN 55455 USA Univ Minnesota Minneapolis MN USA 55455 l Inst, Minneapolis, MN 55455 USA Univ Minnesota, Dept Chem Engn & Mat Sci, Minneapolis, MN 55455 USA Univ Minnesota Minneapolis MN USA 55455 at Sci, Minneapolis, MN 55455 USA
Titolo Testata:
MOLECULAR MICROBIOLOGY
fascicolo: 3, volume: 40, anno: 2001,
pagine: 530 - 541
SICI:
0950-382X(200105)40:3<530:AOTASE>2.0.ZU;2-D
Fonte:
ISI
Lingua:
ENG
Soggetto:
STREPTOMYCES-COELICOLOR A3(2); DNA-BINDING PROTEIN; ANTIBIOTIC PRODUCTION; ESCHERICHIA-COLI; GENE-EXPRESSION; CLAVULIGERUS; REPORTER; LIVIDANS; CLUSTER; CLONING;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
31
Recensione:
Indirizzi per estratti:
Indirizzo: Sherman, DH Univ Minnesota, Dept Microbiol, Mayo Mail Code 196,420 Delaware St SE, Minneapolis, MN 55455 USA Univ Minnesota Mayo Mail Code 196,420 Delaware St SE Minneapolis MN USA 55455
Citazione:
Y.S. Kyung et al., "Analysis of temporal and spatial expression of the CcaR regulatory elementin the cephamycin C biosynthetic pathway using green fluorescent protein", MOL MICROB, 40(3), 2001, pp. 530-541

Abstract

The DNA-binding capability of a key secondary metabolite regulatory element (CcaR) in the Streptomyces clavuligerus cephamycin C pathway was investigated by gel mobility retardation and DNase I footprinting analysis. These results revealed that CcaR specifically binds to the promoter region of the lysine-epsilon -aminotransferase gene (lat). Green fluorescent protein (GFP) was subsequently used as a reporter to analyse in vivo expression of CcaR. The corresponding isogenic strain containing ccaR::gfp in the chromosome produced cephamycin C at levels similar to those of wild-type S. clavuligerus. Confocal laser scanning microscopy revealed that expression of CcaR in liquid culture was temporally dynamic and spatially heterogeneous in S. clavuligerus mycelia. The highly fluorescent seed culture mycelia quickly lostfluorescence upon inoculation into fresh culture medium. The characteristic green colour reappeared in a small portion of mycelia during mid-exponential growth phase. As the culture aged, the population expressing CcaR expanded, and the expression level increased. This was followed by a reduction in the CcaR-expressing population towards the end of the culture period. During peak expression, CcaR was distributed uniformly in mycelia, but became localized distal to the chromosome when the culture entered stationary phase. In solid phase analysis, abundant CcaR expression was evident in the substrate mycelia, but was completely absent in aerial hyphae. These results show regulatory linkage between ccaR and lat, whose expression profile showed a similar spatial decoupling between morphogenesis and antibiotic production. In addition, visualizing CcaR within S. clavuligerus mycelia demonstrates a distinct pattern of localization over the course of physiological differentiation.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 03/12/20 alle ore 11:13:51