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Titolo:
Transfection of eastern oyster (Crassotrea virginica) embryos
Autore:
Buchanan, JT; Nickens, AD; Cooper, RK; Tiersch, TR;
Indirizzi:
Louisiana State Univ, Dept Oceanog & Coastal Sci, Baton Rouge, LA 70820 USA Louisiana State Univ Baton Rouge LA USA 70820 , Baton Rouge, LA 70820 USA Louisiana State Univ, Ctr Agr, Louisiana Agr Expt Stn, Aquaculture Res Stn, Baton Rouge, LA 70803 USA Louisiana State Univ Baton Rouge LA USA 70803 , Baton Rouge, LA 70803 USA Louisiana State Univ, Ctr Agr, Louisiana Agr Expt Stn, Dept Vet Sci, BatonRouge, LA 70803 USA Louisiana State Univ Baton Rouge LA USA 70803 i, BatonRouge, LA 70803 USA
Titolo Testata:
MARINE BIOTECHNOLOGY
fascicolo: 4, volume: 3, anno: 2001,
pagine: 322 - 335
SICI:
1436-2228(200107/08)3:4<322:TOEO(V>2.0.ZU;2-E
Fonte:
ISI
Lingua:
ENG
Soggetto:
STARBURST POLYAMIDOAMINE DENDRIMERS; NEOMYCIN PHOSPHOTRANSFERASE GENE; GREEN FLUORESCENT PROTEIN; LUCIFERASE REPORTER GENE; CRASSOSTREA-GIGAS; TRANSIENT EXPRESSION; EUKARYOTIC CELLS; TRANSGENIC FISH; EFFICIENT TRANSFER; NEO GENE;
Keywords:
Crassostrea virginica; gene transfer; antibiotic resistance; green fluorescent protein; transgenic bivalve;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Agriculture,Biology & Environmental Sciences
Citazioni:
61
Recensione:
Indirizzi per estratti:
Indirizzo: Tiersch, TR Louisiana State Univ, Aquaculture Res Stn, 2410 Ben Hur Rd, Baton Rouge, LA 70820 USA Louisiana State Univ 2410 Ben Hur Rd Baton Rouge LAUSA 70820
Citazione:
J.T. Buchanan et al., "Transfection of eastern oyster (Crassotrea virginica) embryos", MAR BIOTEC, 3(4), 2001, pp. 322-335

Abstract

There is a need for research in disease resistance and microbial elimination in the eastern oyster Crassosostrea virginica. Gene transfer may lead toadvances in this area, and a means of selecting transfected larvae would be useful. We transfected 3-hour-postfertilization embryos with the bacterial gene aminoglycoside phosphotransferase II (neo(r)), which confers resistance to neomycin and related antibiotics such as G418. The antibiotic G418 was examined as a potential selective agent. A neutral red assay was used todetermine survival after 48 hours of exposure to various concentrations ofG418 (0-4 mg/ml). We examined the effects of electroporation and chemically mediated transfection of 3-hour- postfertilization embryos on survival tostraight-hinge larvae. DNA alone was found to have no effect on survival (P > .05). For electroporation we found that increased voltage and pulse duration decreased survival (P < .05). Chemically mediated transfection did not significantly affect survival (P = .5172). Transgenic larvae were identified after electroporation and chemically mediated transfection. These larvae were reared for 24 hours and exposed to G418 at 0.3 mg/ml for 48 hours. Significant differences in survival between transfected and nontransfected larvae were detected for electroporation (P = .0147) and chemically mediatedtransfection (P = .037). Gene transfer was also confirmed with polymerase chain reaction and observation of expression of green fluorescent protein. This Study documents the first successful insertion and expression of foreign DNA in eastern oyster larvae.

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Documento generato il 29/02/20 alle ore 09:06:16