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Titolo:
Production of transgenic live-bearing fish and crustaceans with replication-defective pantropic retroviral vectors
Autore:
Sarmasik, A; Chun, CZ; Jang, IK; Lu, JK; Chen, TT;
Indirizzi:
Univ Connecticut, Ctr Biotechnol, Storrs, CT 06269 USA Univ Connecticut Storrs CT USA 06269 Ctr Biotechnol, Storrs, CT 06269 USA Univ Connecticut, Dept Mol & Cell Biol, Storrs, CT 06269 USA Univ Connecticut Storrs CT USA 06269 ol & Cell Biol, Storrs, CT 06269 USA
Titolo Testata:
MARINE BIOTECHNOLOGY
, volume: 3, anno: 2001, supplemento:, 1
pagine: S177 - S184
SICI:
1436-2228(2001)3:<S177:POTLFA>2.0.ZU;2-M
Fonte:
ISI
Lingua:
ENG
Soggetto:
GENE-TRANSFER; HIGH-TITER; INTEGRATION; EXPRESSION; TRANSMISSION; HEPATOCYTES; CELLS;
Keywords:
replication-defective pantropic retroviral vectors; live-bearers; crustaceans; gene transfer; transgenic animal;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Agriculture,Biology & Environmental Sciences
Citazioni:
18
Recensione:
Indirizzi per estratti:
Indirizzo: Chen, TT Univ Connecticut, Ctr Biotechnol, 184 Auditorium Rd,U-149, Storrs, CT 06269 USA Univ Connecticut 184 Auditorium Rd,U-149 Storrs CT USA 06269 USA
Citazione:
A. Sarmasik et al., "Production of transgenic live-bearing fish and crustaceans with replication-defective pantropic retroviral vectors", MAR BIOTEC, 3, 2001, pp. S177-S184

Abstract

Transgenic fish have been routinely produced by microinjecting or electroporating foreign DNA into one-cell stage embryos or unfertilized eggs. Whileboth techniques are effective in producing transgenic fish species from which unfertilized or newly fertilized eggs can be easily obtained, these techniques are not applicable to live-bearing fish and many crustacean specieswhere unfertilized or newly fertilized eggs are not readily available. In this paper, we describe a new method of introducing foreign DNA into the live-bearing fish, Poeciliposis lucida, and crayfish, Procambarus clarkii, bydirectly transforming the immature ovary or testis of these animals with replication-defective pantropic retroviral vectors carrying a reporter gene (neo(R)). A significant fraction of the progeny derived from these treated animals contains the neo(R) reporter gene, determined by a PCR-based assay. The PCR-positive individuals were crossed with nontransgenic individuals, and about 50% of the resulting progeny carried the transgene, suggesting that the F-1 animals are germline transgenic. Integration of the transgenes was confirmed by detecting the junction fragments of the genomic DNA associated with transgene constructs. The expression of reporter genes was detected by reverse transcription (RT) PCR assay. These results showed that foreign genes could be reproducibly transferred into live-bearing Fish and crustaceans by directly transforming the immature gonads with replication-defective pantropic retroviral vectors.

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Documento generato il 19/02/20 alle ore 15:04:22