RESUMEN
A peroxiredoxin6 (PRDX6)-like gene segment has been found in up-regulated cDNA libraries of earthworm Eisenia fetida exposed to benzo[a]pyrene(BaP). Analysis with basic local alignment search tool (BLAST) and phylogenetic analysis revealed that the gene segment contains a characteristic motif which was encoded by Cys, illustrating that this gene segment belongs to PRDX. To verify the response of PRDX in E. feitida exposed to poly aromatic hydrocarbons (PAHs), four-week pollution experiments were conducted following the methods recommended by the Organization for Economic Co-operation and Development (OECD). The results showed that the expression of PRDX was up-regulated in earthworm exposed to 1.0 mg x kg(-1) Pyrene and BaP. It was indicated that PRDX in E. fetida was the potential molecular biomarkers of antioxidative stress caused by soil contamination.
Asunto(s)
Monitoreo del Ambiente/métodos , Oligoquetos/genética , Peroxiredoxina VI/metabolismo , Hidrocarburos Policíclicos Aromáticos/toxicidad , Contaminantes del Suelo/toxicidad , Secuencia de Aminoácidos , Animales , Benzo(a)pireno/análisis , Benzo(a)pireno/toxicidad , Datos de Secuencia Molecular , Oligoquetos/efectos de los fármacos , Oligoquetos/fisiología , Peroxiredoxina VI/genética , Hidrocarburos Policíclicos Aromáticos/análisis , Contaminantes del Suelo/análisis , Estrés Fisiológico/fisiologíaRESUMEN
The earthworm Eisenia fetida's benzo [a] pyrene (BaP) exposure experiments were carried out in artificial soil according to ISO 11268-1:1993. And then the upregulated and downregulated subtractive cDNA libraries were constructed by Clontech PCR-Select cDNA Subtration Kit. From the BaP exposure upregulated subtractive cDNA library, several cDNA segments matched mitochondrion-encoded genes were found, including cytochrome c oxidase subunit I (CO I), subunit II (CO II), subunit Ill (CO III), NADH dehydrogenase subunit 1 (NDH1), and ATP synthase subunit 6. The result indicated BaP and the subsequent oxidative stress disturbed the expression of mitochondrion-encoded genes, and this was potential biomarker for oxidative stress following xenobiotic exposure.
Asunto(s)
Benzo(a)pireno/toxicidad , Genes Mitocondriales/efectos de los fármacos , Oligoquetos/efectos de los fármacos , Contaminantes del Suelo/toxicidad , Secuencia de Aminoácidos , Animales , ADN Mitocondrial/química , ADN Mitocondrial/genética , Complejo IV de Transporte de Electrones/genética , Expresión Génica/efectos de los fármacos , Genes Mitocondriales/genética , ATPasas de Translocación de Protón Mitocondriales/genética , Datos de Secuencia Molecular , NADH Deshidrogenasa/genética , Oligoquetos/genética , Análisis de Secuencia de ADN , Homología de Secuencia de AminoácidoRESUMEN
To search for the molecular biomarkers of sub-lethal polycyclic aromatic hydrocarbons (PAHs)-contamination of soil, the subtractive cDNA libraries of earthworm Eisenia fetida exposed to benzo(a)pyrene (BaP) in artificial soil were constructed by suppression subtractive hybridization. After sequencing and analyzing with basic local alignment search tool (BLAST), two clones matching heat shock protein 70 k Da (HSP70) and one clone matching heat shock protein 90 k Da (HSP90) were isolated from the up-regulated library, and subsequently, the up-regulation of HSP70 and HSP90 was verified by real-time PCR in E. fetida exposed to 0.1 mg x kg(-1) and 1.0 mg x kg(-1) BaP. It was indicated that these two newly identified HSPs in E. fetida were the potential molecular biomarkers for soil contamination monitoring.