Amplificación y clonación de cDNAs de los genes del citochromo P4501A1 y la metalotioneína de Sparus aurata Linnaeus, 1758 y Liza aurata (Risso, 1810) por Race-PCR
2001; American Association for the Advancement of Science; Linguagem: Inglês
ISSN
1095-9203
AutoresM. Cousinou, G. Dorado, Juan López‐Barea,
Tópico(s)Environmental Toxicology and Ecotoxicology
ResumoThe biotransformation of xenobiotics found in marine ecosystems is catalysed by inducible systems, a property that makes them useful as early-warning biomarkers of environmental pollution. We have focused our study on cytochrome P4501A1 ( CYP 1A1) as a phase I response against organic aromatic xenobiotics, and metallothionein ( MT ), which reflects pollution by transition metals. The high homology existing between the sequences of both genes, already cloned in different fish species, enabled us to design degenerate oligos to amplify by RT-PCR specific sequences of CYP 1A1 and MT genes in two teleost fish species of the Spanish South Atlantic littoral, the gilthead seabream, Sparus aurata Linnaeus, 1758, and the grey mullet, Liza aurata (Risso 1810). To this end, the expression of both genes was previously induced by intraperitoneal injection with Aroclor 1254 ( CYP 1A1) and CdCl2 ( MT ). The specific sequences for both genes amplified by RT-PCR with degenerate oligos were subsequently used to design new, specific oligos to obtain by Race-PCR (Rapid Amplification of cDNA Ends) the complete cDNAs coding for both genes in both fish species, which were subsequently cloned and sequenced. The coding sequences and the corresponding proteins were compared with those already obtained in other fish species. We are currently developing homologous probes for mRNA quantification using molecular biology techniques in both fish species, to be employed as molecular biomarkers of pollution in the Spanish South Atlantic littoral.
Referência(s)