Int J Phytoremediation. 2014;16(7-12):937-46. doi: 10.1080/15226514.2013.810575.
Genetically modified plants can serve as an efficient tool for remediation of diverse dangerous pollutants of the environment such as pesticides, heavy metals, explosives and persistent organic compounds. Transgenic lines of Nicotiana tabacum containing bacterial bphC gene from the degradation pathway of polychlorinated biphenyls (PCBs) were tested. The product of the bphC gene - enzyme 2,3-dihydroxybiphenyl-1,2-dioxygenase is responsible for cleaving of the biphenyl ring. The presence of bphC gene in transgenic plants was detected on DNA, RNA and protein level. The expression of the bphC/His gene was verified afterpurification of the enzyme from plants by affinity chromatography followed by a Western blot and immunochemical assay. The enzyme activity of isolated protein was detected. Efficient transformation of 2,3-DHB by transgenic plants was achieved and the lines also exhibited high production of biomass. The transgenic plants were more tolerant to the commercial PCBs mixture Delor 103 than non-transgenic tobacco. And finally, the higher decrease of total PCB content and especially congener 28 in real contaminated soil from a dumpsite was determined after cultivation of transgenic plant in comparison with nontransgenic tobacco. The substrate specificity of transgenic plants was the same as substrate specificity of BphC enzyme.
转基因植物可作为修复环境中多种危险污染物的有效工具,如农药、重金属、爆炸物和持久性有机化合物。本文测试了含有来自多氯联苯(PCBs)降解途径的细菌 bphC 基因的烟草转基因系。bphC 基因的产物 - 酶 2,3-二羟基联苯-1,2-双加氧酶负责裂解联苯环。在 DNA、RNA 和蛋白质水平上检测到转基因植物中 bphC 基因的存在。通过亲和层析从植物中纯化酶后,通过 Western blot 和免疫化学分析验证了 bphC/His 基因的表达。检测到分离蛋白的酶活性。成功地实现了 2,3-DHB 的有效转化,并且这些系也表现出高生物质产量。与非转基因烟草相比,转基因植物对商业 PCB 混合物 Delor 103 的耐受性更高。最后,与非转基因烟草相比,在受污染的垃圾场实际污染土壤中种植转基因植物后,确定总 PCB 含量特别是同系物 28 的含量有更高的降低。转基因植物的底物特异性与 BphC 酶的底物特异性相同。