마이크로어레이를 이용한 조피볼락(Sebastes schlegeli)에서 벤조피렌에 의해 유도되는 유전자의 스크리닝

Abstract

Benzo(a)pyrene (BaP) is a representative ecotoxicant in marine environment and a model compound of polycyclic aromatic hydrocarbons(PAHs), which has an ability to bioaccumulate in aquatic organisms. This study aimed to identify molecular biomarkers suitable for assessing environmental pollution using a microarray technique. Specifically, we examined the effects of BaP on gene expressions in the rockfish,Sebastes schlegeli. We constructed the subtractive cDNA library with liver RNA from BaP-exposed fish and non-exposed control fish and obtained 10,000 candidate clones which gene expressions were altered by BaP-exposure. We determined BaP-responsive genes using ahigh-density microarray of 10,000 candidate genes of mentioned above. To determine temporal changes in gene expression post-BaPtreatment, we compared transcriptome levels between non-exposed and BaP-exposed fishes. Statistical analysis showed that approximately 400 genes are significantly induced or reduced by BaP treatment (2 uM). Molecular characterizations including nucleotide sequence, full length gene cloning, and estimation of the functions of these genes are being processed. Many methods have been developed to monitormarine environmental status, which depend on quantifying the levels of the toxic components in polluted seawater or on ecologicalaccessing, such as species diversity or richness. However, those methods could not provide information on physiological or genetic changes induced by such environmental stresses. Comparing with the conventional methods, our data will propose that these BaP-responsible genes can be useful for biological risk assessment of PAHs on marine organism at molecular level.