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Titolo:
Inhibition of Trypanosoma brucei gene expression by RNA interference usingan integratable vector with opposing T7 promoters
Autore:
Wang, ZF; Morris, JC; Drew, ME; Englund, PT;
Indirizzi:
Johns Hopkins Univ, Sch Med, Dept Biol Chem, Baltimore, MD 21205 USA JohnsHopkins Univ Baltimore MD USA 21205 l Chem, Baltimore, MD 21205 USA
Titolo Testata:
JOURNAL OF BIOLOGICAL CHEMISTRY
fascicolo: 51, volume: 275, anno: 2000,
pagine: 40174 - 40179
SICI:
0021-9258(200012)275:51<40174:IOTBGE>2.0.ZU;2-9
Fonte:
ISI
Lingua:
ENG
Soggetto:
DOUBLE-STRANDED-RNA; CRITHIDIA-FASCICULATA; C-ELEGANS; NEUROSPORA; POLYMERASE; PROTEIN; REPLICATION; HELICASE; CLONING; DSRNA;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
33
Recensione:
Indirizzi per estratti:
Indirizzo: Englund, PT Johns Hopkins Univ, Sch Med, Dept Biol Chem, 725 N Wolfe St, Baltimore, MD21205 USA Johns Hopkins Univ 725 N Wolfe St Baltimore MD USA 21205 5 USA
Citazione:
Z.F. Wang et al., "Inhibition of Trypanosoma brucei gene expression by RNA interference usingan integratable vector with opposing T7 promoters", J BIOL CHEM, 275(51), 2000, pp. 40174-40179

Abstract

RNA interference is a powerful method for inhibition of gene expression inTrypanosoma brucei (Ngo, H., Tschudi, C,, Gull, K,, and Ullu, E. (1998) Proc. Natl. Acad. Sci. U. S. A. 95, 14687-14692), Here we describe a vector (pZJM) for in vivo tetracycline-inducible synthesis of double-stranded RNA (dsRNA) in stably transformed cells. The dsRNA is synthesized from opposing T7 promoters. We tested the vector with genes involved in processes such askinetoplast DNA replication, mitochondrial mRNA synthesis, glycosyl phosphatidylinositol biosynthesis, glycosome biogenesis, and polyamine biosynthesis, In most cases the induction of dsRNA caused specific and dramatic loss of the appropriate mRNA and in many cases there was growth inhibition or cell death. One striking phenotype was the loss of kinetoplast DNA after interference with expression of a topoisomerase II. The gene being analyzed by this procedure need not even be fully sequenced. In fact, many of the geneswe tested were derived from partial sequences in the T. brucei genome database that were identified by homology with known proteins. It takes as little as 3 weeks from identification of a gene sequence in the data base to the appearance of a phenotype.

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Documento generato il 28/09/20 alle ore 02:20:16