拟南芥多药物和有毒化合物排出家族属次级转运蛋白家族,此类转运蛋白与解毒内源的次生代谢物和外源的有毒化合物有关。通过PCR的方法从拟南芥基因组中扩增到该家族成员DTX12的启动子序列,构建双元载体pBI101.2-ProDTX12-GUS,通过农杆菌介导的方法转化拟南芥,然后对转基因植株用GUS底物进行组织化学显色分析。同时,通过半定量RT-PCR的方法,进一步验证了DTX12在不同组织中的表达情况。结果表明该基因在成熟的花器官的花药中和幼苗的根尖特异表达,另外,在子叶的尖端也有少量的表达。由于DTX12编码的是一个具有转运有毒化合物功能的蛋白,推测其功能可能是转运与细胞分裂或生长有关的次生代谢物。
The multidrug and toxic compound extrusion family belongs to the secondary transporter family. Members of the MATE family are found to be involved in the detoxification of endogenous secondary metabolites and xenobiotics. In this paper, we amplified the promoter sequence of DTX12, one of the MATE family genes, by PCR technique , and then constructed it into binary plasmid pBI101.2, fusing to GUS gene, subsequently transformed into Arabidopsis by Agrobacterium mediated method, finally analyzed the histochemical GUS staining result. At the same time, we confirmed the expression pattern of DXT12 again by semi-quantitative RT-PCR. The result shows that this gene expresses specifically in the anthers of mature flowers, root tips and the cotyledon tips of seedling. As a multidrug and toxic compound transporter, DXT12 is presumed to be a transporter of secondary metabolites involved in cell division and elongation.