Novakova Martina, Mackova Martina, Antosova Zuzana, Viktorova Jitka, Szekeres Miklos, Demnerova Katerina, Macek Tomas
ICT Prague, Faculty of Food and Biochemical Technology, Department of Biochemistry and Microbiology, Prague, Czech Republic.
Bioeng Bugs. 2010 Nov-Dec;1(6):419-23. doi: 10.4161/bbug.1.6.12723.
The aim of this work was to construct transgenic plants with increased capabilities to degrade organic pollutants, such as polychlorinated biphenyls. The environmentally important gene of bacterial dioxygenase, the bphC gene, was chosen to clone into a plant of Nicotiana tabacum. The chosen bphC gene encodes 2,3-dihydroxybiphenyl-1,2-dioxygenase, which cleaves the aromatic ring of dihydroxybiphenyl, and we cloned it in fusion with the gene for β-glucuronidase (GUS), luciferase (LUC) or with a histidine tail. Several genetic constructs were designed and prepared and the possible expression of desired proteins in tobacco plants was studied by transient expression. We used genetic constructs successfully expressing dioxygenase's genes we used for preparation of transgenic tobacco plants by agrobacterial infection. The presence of transgenic DNA , mRNA and protein was determined in parental and the first filial generation of transgenic plants with the bphC gene. Properties of prepared transgenic plants will be further studied.
这项工作的目的是构建具有增强降解有机污染物(如多氯联苯)能力的转基因植物。选择细菌双加氧酶的环境重要基因bphC基因,将其克隆到烟草植株中。所选的bphC基因编码2,3 - 二羟基联苯 - 1,2 - 双加氧酶,该酶可切割二羟基联苯的芳香环,我们将其与β - 葡萄糖醛酸酶(GUS)、荧光素酶(LUC)基因或带有组氨酸尾巴的基因融合进行克隆。设计并制备了几种基因构建体,并通过瞬时表达研究了所需蛋白质在烟草植株中的可能表达情况。我们使用成功表达双加氧酶基因的基因构建体,通过农杆菌感染制备转基因烟草植株。在含有bphC基因的转基因植物的亲代和第一代子代中测定转基因DNA、mRNA和蛋白质的存在情况。所制备的转基因植物的特性将进一步研究。