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Titolo:
Development of a noninvasive reporter system for gene expression in Porphyromonas gingivalis
Autore:
Liu, Y; Abaibou, H; Fletcher, HM;
Indirizzi:
Loma Linda Univ, Sch Med, Dept Microbiol & Mol Genet, Loma Linda, CA 92350USA Loma Linda Univ Loma Linda CA USA 92350 ol Genet, Loma Linda, CA 92350USA
Titolo Testata:
PLASMID
fascicolo: 3, volume: 44, anno: 2000,
pagine: 250 - 261
SICI:
0147-619X(200011)44:3<250:DOANRS>2.0.ZU;2-B
Fonte:
ISI
Lingua:
ENG
Soggetto:
GREEN FLUORESCENT PROTEIN; ARG-GINGIPAIN; BACTEROIDES; VIRULENCE; CLONING; PRTH;
Keywords:
Porphyromonas; Porphyromonas gingivalis; reporter gene; gene expression; xylosidase; arabinosidase; xa; luciferase;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
28
Recensione:
Indirizzi per estratti:
Indirizzo: Fletcher, HM Loma Linda Univ, Sch Med, Dept Microbiol & Mol Genet, Loma Linda, CA 92350USA Loma Linda Univ Loma Linda CA USA 92350 a Linda, CA 92350USA
Citazione:
Y. Liu et al., "Development of a noninvasive reporter system for gene expression in Porphyromonas gingivalis", PLASMID, 44(3), 2000, pp. 250-261

Abstract

Several reports have supported the association of Porphyromonas gingivaliswith periodontal disease. Genetic studies are vital for understanding the relative importance of virulence factors in this organism. Thus, gene reporters may prove useful for the study of gene expression in this organism. Wehave investigated the use of the green fluorescent protein (GTP), bacterial luciferase, and bifunctional xylosidase/arabinosidase enzyme (XA) as reporters of gene expression in P. gingivalis. Fusion cassettes containing the promoterless tetracycline resistant gene [tetA(A)Q2] and the promoterless gfp, luxAB, or xa gene were placed under the control of the rgpA promoter inP. gingivalis W83 using recombinational allelic exchange. The rgpA gene encodes for an arginine-specific pretense in P. gingivalis. No GFP activity was detected in P. gingivalis isogenic mutants carrying the rgpA::gfp-tetA(Q)2 fusion construct. Luciferase activity in P. gingivalis mutants carrying the rgpA::luxAB-tetA(Q)2 fusion was only detected in the presence of exogenous FMNH2. xa gene expression in P. gingivalis with the rgpA::xa-tetA(Q)2 fusion construct was detected in crude extracts using rho -nitrophenol derivatives as substrate and on agar plates with methylumbelliferyl derivatives under long-wave ultraviolet light. This indicates that both luxAB and xa genes can be used as reporters of gene expression in P, gingivalis. However, only the xa gene can be used as a noninvasive reporter gene. (C) 2000 Academic Press.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 27/11/20 alle ore 13:20:34