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

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...

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Autores principales:	Villalobos,Patricio, Rojas,M. Verónica, Conejeros,Pablo, Marshall,Sergio H.
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 1999
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34581999000200005
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spelling	oai:scielo:S0717-345819990002000052003-08-18Lipopolyamine-mediated transfection of reporter plasmids into a fish cell lineVillalobos,PatricioRojas,M. VerónicaConejeros,PabloMarshall,Sergio H. 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 transgenesisinfo:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.2 n.2 19991999-08-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34581999000200005en10.4067/S0717-34581999000200005
institution	Scielo Chile
collection	Scielo Chile
language	English
description	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
author	Villalobos,Patricio Rojas,M. Verónica Conejeros,Pablo Marshall,Sergio H.
spellingShingle	Villalobos,Patricio Rojas,M. Verónica Conejeros,Pablo Marshall,Sergio H. Lipopolyamine-mediated transfection of reporter plasmids into a fish cell line
author_facet	Villalobos,Patricio Rojas,M. Verónica Conejeros,Pablo Marshall,Sergio H.
author_sort	Villalobos,Patricio
title	Lipopolyamine-mediated transfection of reporter plasmids into a fish cell line
title_short	Lipopolyamine-mediated transfection of reporter plasmids into a fish cell line
title_full	Lipopolyamine-mediated transfection of reporter plasmids into a fish cell line
title_fullStr	Lipopolyamine-mediated transfection of reporter plasmids into a fish cell line
title_full_unstemmed	Lipopolyamine-mediated transfection of reporter plasmids into a fish cell line
title_sort	lipopolyamine-mediated transfection of reporter plasmids into a fish cell line
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	1999
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34581999000200005
work_keys_str_mv	AT villalobospatricio lipopolyaminemediatedtransfectionofreporterplasmidsintoafishcellline AT rojasmveronica lipopolyaminemediatedtransfectionofreporterplasmidsintoafishcellline AT conejerospablo lipopolyaminemediatedtransfectionofreporterplasmidsintoafishcellline AT marshallsergioh lipopolyaminemediatedtransfectionofreporterplasmidsintoafishcellline
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Lipopolyamine-mediated transfection of reporter plasmids into a fish cell line

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