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| Villalobos,Patricio; Rojas,M. Verónica; Conejeros,Pablo; Marshall,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... |
| Tipo: Journal article |
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| Ano: 1999 |
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34581999000200005 |
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| Jorquera,Esteban; Morales,Paz; Tapia,David; Torres,Pamela; Eissler,Yoanna; Espinoza,Juan C; Conejeros,Pablo; Kuznar,Juan. |
| Background: The genomes of several infectious pancreatic necrosis viruses (IPNVs) isolated in Chile were sequenced with a single amplification approach for both segments A and B. The resulting sequences were then used to determine the conservation of the primer-binding regions used in polymerase chain reaction (PCR)-based diagnostic methods proposed in the literature. Thus, the robustness of each technique was studied, particularly the eventual effect of further mutations within the primer-binding sites. Results: On analysis, most methods currently used to detect Chilean IPNV varieties were deemed adequate. However, the primers were designed to be genogroup specific, implying that most detection methods pose some risk of detecting all strains prevalent in... |
| Tipo: Journal article |
Palavras-chave: IPNV detection; Mismatch's Tm analysis; QPCR. |
| Ano: 2016 |
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582016000200005 |
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