首页> 外国专利> METHOD FOR DETERMINING GENOTOXICITY OF XENOBIOTICS BASED ON ANALYSIS OF DAMAGE TO MITOCHONDRIAL DNA OF EARTH BUMBLEBEE (BOMBUS TERRESTRIS)

METHOD FOR DETERMINING GENOTOXICITY OF XENOBIOTICS BASED ON ANALYSIS OF DAMAGE TO MITOCHONDRIAL DNA OF EARTH BUMBLEBEE (BOMBUS TERRESTRIS)

机译：基于对土蜂线粒体DNA损伤分析的异种药遗传毒性测定方法

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FIELD: biotechnology.;SUBSTANCE: invention relates to biotechnology. Method for determining the genotoxicity of xenobiotics based on an analysis of damage to mitochondrial DNA of earth bumblebee (Bombus terrestris). Method comprises feeding ground bumblebees (Bombus terrestris) with the tested xenobiotics throughout the day, sampling of biological material and isolation of DNA from pectoral muscles of the bumblebee, determination of the quantitative level of damage to mitochondrial DNA by real-time PCR at an elongation temperature of 66.1±0.5 °C for 5 minutes using Encyclo-polymerase corresponding to Encyclo-buffer and other standard reaction components with primer pairs having specific sequences, comparison of the quantitative level of damage to the mitochondrial DNA of the experimental and control groups. Higher level of damage in the experimental group is an indicator of the genotoxicity of xenobiotics. Sequences of primers for determining the amount of damage in mitochondrial DNA B. terrestris and the number of its copies:; <TableGroup colSep="1" cols="3" rowSep="1"> <Colspec colName="c1" colWidth="10mm"/> <Colspec colName="c2" colWidth="73mm"/> <Colspec colName="c3" colWidth="75mm"/> <Tbody> <Row> <entry colSep="1" rowSep="1">No.</entry> <entry colSep="1" rowSep="1">Straight</entry> <entry colSep="0" rowSep="1">Back</entry> </Row> <Row> <entry colSep="1" rowSep="1">1</entry> <entry colSep="1" rowSep="1">CCCCAGATATAGCTTTTCCTC</entry> <entry colSep="0" rowSep="1">CCAGGAATTGCATCAACTTT</entry> </Row> <Row> <entry colSep="1" rowSep="1">2</entry> <entry colSep="1" rowSep="1">CTTCAATTCACCTTTAAAACAA</entry> <entry colSep="0" rowSep="1">GTATTACCACGAATTCGATATG</entry> </Row> <Row> <entry colSep="1" rowSep="0">3</entry> <entry colSep="1" rowSep="0">CGCTATTGCTGGCACTAATTT</entry> <entry colSep="0" rowSep="0">AAATTATTCAGAAACAAAATGGAAA</entry> </Row> </Tbody> </TableGroup> </TableFormula> </Table> ;Present invention can be used in testing various xenobiotics, including pesticides, for genotoxicity.;EFFECT: technical result consists in development of highly sensitive, highly reproducible and relatively inexpensive way to determine the genotoxicity of xenobiotics.;1 cl, 2 dwg, 1 ex </span> <span class="z_kbtn z_kbtnclass hoverxs" style="display: none;">展开▼</span> </div> <div class="translation abstracttxt"> <span class="zhankaihshouqi fivelineshidden" id="abstract"> <span>机译：</span>技术领域：本发明涉及生物技术。基于对大黄蜂线粒体DNA损伤的分析来确定异种生物的遗传毒性的方法。该方法包括全天为地面大黄蜂（<I> Bombus terrestris </ I>）饲喂经过测试的异种生物，取样生物材料并从大黄蜂的胸肌中分离DNA，并通过以下方法确定线粒体DNA损伤的定量水平使用对应于Encyclo-buffer和其他标准反应组分的Encyclo-聚合酶和具有特定序列的引物对，在66.1±0.5°C的延伸温度下进行5分钟实时PCR，比较对线粒体DNA损伤的定量水平实验组和对照组。实验组中较高水平的损伤是异生物素遗传毒性的指标。用于确定线粒体DNA IB中的损伤量的引物序列。 terrestris </ I>及其副本数： <表格> <TableFormula> <标题/> <TableGroup colSep =“ 1” cols =“ 3” rowSep =“ 1”> <Colspec colName =“ c1” colWidth =“ 10mm” /> <Colspec colName =“ c2” colWidth =“ 73mm” /> <Colspec colName =“ c3” colWidth =“ 75mm” /> <身体> <行> <entry colSep =“ 1” rowSep =“ 1”>否。</ entry> <entry colSep =“ 1” rowSep =“ 1”>直接</ entry> <entry colSep =“ 0” rowSep =“ 1”>返回</ entry> </ Row> <行> <entry colSep =“ 1” rowSep =“ 1”> 1 </ entry> <entry colSep =“ 1” rowSep =“ 1”> CCCCAGATATAGCTTTTCCTC </ entry> <entry colSep =“ 0” rowSep =“ 1”> CCAGGAATTGCATCAACTTT </ entry> </ Row> <行> <entry colSep =“ 1” rowSep =“ 1”> 2 </ entry> <entry colSep =“ 1” rowSep =“ 1”> CTTCAATTCACCTTTAAAACAA </ entry> <entry colSep =“ 0” rowSep =“ 1”> GTATTACCACGAATTCGATATG </ entry> </ Row> <行> <entry colSep =“ 1” rowSep =“ 0”> 3 </ entry> <entry colSep =“ 1” rowSep =“ 0”> CGCTATTGCTGGCACTAATTT </ entry> <entry colSep =“ 0” rowSep =“ 0”> AAATTATTCAGAAACAAAATGGAAA </ entry> </ Row> </ Tbody> </ TableGroup> </ TableFormula> </ Table> ;本发明可用于测试包括农药在内的各种异源生物的遗传毒性。效果：技术成果在于开发了一种高度灵敏，高度可重复且相对廉价的测定异源生物遗传毒性的方法。1 cl，2 dwg，1前 </span> <span class="z_kbtn z_kbtnclass hoverxs" style="display: none;">展开▼</span> </div> </div> <div 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