Construction and characterization of Japanese medaka (Oryzias latipes) hepatic cDNA library and its implementation to biomarker screening in aquatic toxicology
- Construction and characterization of Japanese medaka (Oryzias latipes) hepatic cDNA library and its implementation to biomarker screening in aquatic toxicology
- Pham, Chi Hoa; 박경서; 김병찬; 김한나; 구만복
- cDNA library; Aquatic toxicology; Medaka; Biomaker; Real time RT-PCR
- Issue Date
- Aquatic toxicology
- VOL 105, NO 3-4, 569-575
- To strengthen the toxicogenomic study, we constructed a library of hepatic cDNA from Japanese medaka
under influence of specific chemical mediated stress responses. Gene expression profile analysis of the
cDNA microarrays followed by real time RT-PCR assay were conducted to screen particular biomarkers
for 17-beta estradiol (E2), nonylphenol (NP) and 2-chlorophenol (2CP). Information of 1509 high-quality
ESTs including 260 new ESTs was added onto GenBank and dbEST. The ESTs were clustered and assembled
into 159 contigs and 372 singletons. Among them, 128 contigs and 163 singletons (54.8%) were functionally
characterized and 13 UniESTs (2.5%) were hypothetical proteins. Ontology analysis resulting in 282
UniESTs which involved with 2102 GOs and 93 sequences associated with 116 enzyme codes. For each
test chemical, two specific biomarkers were selected from the gene expression profiling of microarrays.
The expression patterns of the marker genes in real time PCR analysis were consistent with the regulated
gene expression patterns in microarrays. The tentative biomarkers showed unique gene expression
patterns depending on chemical concentration(s) and exposure duration in real time RT-PCR analysis.
The analysis accomplished of the hepatic cDNA library and its information added to genetic and genomic
resources could be sufficiently valuable specifically for aquatic toxicity studies.
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