【摘要】 目的:为用基因工程的方法人工表达芳香烃受体(AhR)及芳香烃受体核转运蛋白(ARNT),构建了融合蛋白的表达载体。方法:抽提C57BL/6J小鼠肝脏总RNA后采用RT-PCR的方法扩增得到芳香烃受体及其核转运蛋白的全长序列。在DNA测序确定其正确性之后,连入表达载体并转化大肠杆菌DH5α感受态细胞,经氨苄青霉素筛选阳性克隆,提取质粒进行酶切分析。结果:成功构建了芳香烃受体及其核转运蛋白的重组表达载体。结论:载体构建成功是基因工程制备芳香烃受体及其核转运蛋白的前提,并为进行二恶英毒性机制及生物学检测方法的研究奠定了基础。更多还原

【Abstract】 Objective: In order to get the proteins of AhR and ARNT by genetic engineering,prokaryotic expression vectors were constructed.Methods: The genes encoding AhR and ARNT were amplified by RT-PCR using the total RNA purified from the liver of C57BL/6J mice.The DNA fragments were ligated into expression vector after the fidelity of them were determined by DNA sequence analysis.Then they were transferred into Ecoli.DH5α competent cells.The positive clones screened by ampicillin were validated by digestion of restrictive endonuclease.Results: The reconstructive expression vectors of AhR and ARNT were successfully established.Conclusion: The construction of expression vectors of AhR and ARNT is the premise of acquisition of them through genetic engineering.It also can provide the basis for establishment of dioxin bioassay system and related research on its toxic mechanism.更多还原