In vitro check for this species in other resources transfection , Reporter gene expression"/>
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Electronic Journal of Biotechnology
Universidad Católica de Valparaíso
ISSN: 0717-3458
Vol. 2, No. 2, 1999, pp. 89-98
Bioline Code: ej99012
Full paper language: English
Document type: Research Article
Document available free of charge

Electronic Journal of Biotechnology, Vol. 2, No. 2, 1999, pp. 89-98

 en Lipopolyamine-mediated transfection of reporter plasmids into a fish cell line
Villalobos, Patricio; Rojas, M. Verónica; Conejeros, Pablo & Marshall, Sergio H.

Abstract

Conditions have been optimised to transfect the fish cell line CHSE-214 to measure expression, maintenance and putative chromosomal integration of the reporter gene LUC, spliced into two versions of an expression vector. The first is pCMVL, and the second p103, a novel pCMVL-derived plasmid to which a highly conserved tandem repeat from the salmon genome was added in an inverted configuration flanking the LUC gene to promote its chromosomal integration. A minimal ratio of one to one, lipopolyamine carrier to plasmid DNA, was enough to efficiently transfect the cell line to follow the fate of target DNAs up to five cell passages. In this time-span we demonstrated the maintenance of the foreign DNA in the cells, the concomitant expression of the reporter gene, and a higher stability of p103 over the control plasmid which might suggest a higher potential for integration. Thus, we define an efficient model system for future in vitro evaluation of potential target genes of commercial interest for fish transgenesis.

Keywords
Chromosomal integration , DNA maintenance , Fish cell line , In vitro check for this species in other resources transfection , Reporter gene expression

 
© 1999 by Universidad Católica de Valparaíso -- Chile
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