苯并[a]芘对赤子爱胜蚓线粒体编码基因表达影响研究

Impact of Benzo[a]pyrene the Expression of Mitochondrion-encoded Genes in the Earthworm Eisenia fetida

根据ISO11268-1:1993方法,完成了土壤中苯并[a]芘对赤子爱胜蚓(Eisenia fetida)的人工污染模拟实验.并采用Clontech PCR-SelectTMcDNASubtration Kit构建了污染组和对照组蚯蚓之间的抑制消减文库.经测序和比对分析,在苯并[a]芘诱导上调文库中发现5个线粒体DNA(mtDNA)编码的基因,且在上调文库中具有较高的频率.它们分别为:细胞色素c氧化酶亚基Ⅰ、Ⅱ、Ⅲ,NADH氢脱氢酶亚基1,ATP合成酶亚基.研究表明,土壤中苯并[a]芘污染改变了蚯蚓线粒体基因组的表达水平,线粒体基因组表达异常是多环芳烃污染胁迫的重要分子生物标记物.

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 eDNA libraries were constructed by Clontech PCR-SelectTM eDNA Subtration Kit. From the BaP exposure upregulated subtractive eDNA library, several eDNA segments matched mitochondrion-encoded genes were found, including cytochrome c oxidase subunit Ⅰ （ CO Ⅰ）, subunit Ⅱ （ CO Ⅱ ）, subunit Ⅲ（ 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.