在前期建立的苯并[a]芘(benzo[a]pyrene,BaP)诱导上调表达的蚯蚓cDNA文库中,发现一种过氧化物还原蛋白peroxiredoxin6(PRDX6)基因片段,并提交到GenBank.Blastx比对与基因进化分析结果表明,该序列与已知PRDX6的最高相似率为86%,期望值为4E-30.序列分析表明,该序列具有PRDX6基因含Cys编码的一个特征motif,表明该序列属于PRDX6基因.为进一步验证蚯蚓PRDX6基因对多环芳烃污染的响应情况,使用OECD推荐方法,进行了土壤菲、芘、荧蒽和苯并芘污染对蚯蚓的毒性响应的试验(周期28 d),并采用定量PCR方法,检测了各实验组蚯蚓PRDX6基因的表达差异.结果发现,1.0 mg·kg-1的芘和苯并[a]芘均可显著上调蚯蚓PRDX6基因的表达,表明PRDX6基因可作为土壤污染引起蚯蚓抗氧化应激检测的分子标记.
A peroxiredoxin6 (PRDX6)-like gene segment has been found in up-regulated eDNA 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-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.