Marine Biotechnology Molecular Biology and Genetics
EJB Electronic Journal of Biotechnology ISSN: 0717-3458 Vol.2 No.2, Issue of August 15, 1999.
© 1999 by Universidad Católica de Valparaíso -- Chile Received March 3 , 1999 / Accepted April 16 , 1999
RESEARCH ARTICLE

Lipopolyamine-mediated transfection of reporter plasmids into a fish cell line

Patricio Villalobos
Laboratorio de Genética Molecular, Instituto de Biología, Facultad de Ciencias Básicas y Matemáticas,
Universidad Católica de Valparaíso, Avenida Brasil 2950, Valparaíso - Chile
Fax 56 32 596703
E-mail : biaggini@ucv.cl

M. Verónica Rojas
Laboratorio de Genética Molecular, Instituto de Biología, Facultad de Ciencias Básicas y Matemáticas,
Universidad Católica de Valparaíso, Avenida Brasil 2950, Valparaíso - Chile
Fax 56 32 596703
E-mail : mvrojas@ucv.cl

Pablo Conejeros
Laboratorio de Genética Molecular, Instituto de Biología, Facultad de Ciencias Básicas y Matemáticas,
Universidad Católica de Valparaíso, Avenida Brasil 2950, Valparaíso - Chile
Fax 56 32 596703


Sergio H. Marshall *
Laboratorio de Genética Molecular, Instituto de Biología, Facultad de Ciencias Básicas y Matemáticas,
Universidad Católica de Valparaíso, Avenida Brasil 2950, Valparaíso - Chile
Fax 56 32 273420
E-mail : vriea@ucv.cl

* Corresponding author

Keywords : Chromosomal integration , DNA maintenance , Fish cell line , In vitro transfection , Reporter gene expression.

Abstract Full Text

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.

Supported by UNESCO / MIRCEN network
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