Faculty of Pharmaceutical Sciences, Chiba University, Yayoi-cho 1-33, 260, Chiba, Japan.
Plant Cell Rep. 1990 Jul;9(3):121-4. doi: 10.1007/BF00232085.
The chimeric neo and gus genes on a mini Ti vector are efficiently transferred into the genome of fox glove (Digitalis purpurea L.) using a binary vector system based on a rootinducing Ri plasmid, pRi15834. The transgenic state of established transformed roots was confirmed by Southern blot analysis and by detection of agropine and mannopine. The expression of the chimeric genes controlled by the promoters from TR 1'-2' genes, nos gene and cauliflower mosaic virus 35S RNA was demonstrated by enzymatic and histochemical assays of neomycin phosphotransferase II and ß-glucuronidase. Enzyme-linked immunosorbent assay (ELISA) was carried out using polyclonal antibody reactable against digitoxin to investigate the production of cardenolides. The results of ELISA indicated that the cardioactive glycosides were highly produced in the green transformed hairy roots.
利用基于发根诱导 Ri 质粒 pRi15834 的二元载体系统，将微型 Ti 载体上的嵌合 neo 和 gus 基因高效地转移到毛地黄（Digitalis purpurea L.）的基因组中。通过 Southern blot 分析和农杆菌碱和曼诺辛的检测，证实了已建立的转化根的转基因状态。通过对新霉素磷酸转移酶 II 和β-葡糖苷酸酶的酶和组织化学分析，证明了由 TR 1'-2'基因、nos 基因和花椰菜花叶病毒 35S RNA 启动子控制的嵌合基因的表达。利用可与地高辛反应的多克隆抗体进行酶联免疫吸附测定（ELISA），以研究强心苷的产生。ELISA 的结果表明，在绿色转化的毛状根中高度产生了强心糖苷。
