Hain R, Bieseler B, Kindl H, Schröder G, Stöcker R
Bayer AG, PF-E/Institut für Biotechnologie, Leverkusen-Bayerwerk, FRG.
Plant Mol Biol. 1990 Aug;15(2):325-35. doi: 10.1007/BF00036918.
A gene from groundnut (Arachis hypogaea) coding for stilbene synthase was transferred together with a chimaeric kanamycin resistance gene. It was found to be rapidly expressed after induction with UV light and elicitor in tobacco cells (Nicotiana tabacum). Comparative studies of stilbene synthase mRNA synthesis in groundnut and transgenic tobacco suspension cultures revealed the same kinetics of gene expression. Stilbene synthase specific mRNA was detectable 30 minutes after elicitor induction and 10 minutes after UV irradiation. The maximum of mRNA accumulation was between 2 and 8 hours post induction. 24 hours after induction stilbene synthase mRNA accumulation ceased. Furthermore, in transgenic tobacco plants, the gene was found to be inducible in sterile roots, stems and leaves. Stilbene synthase was demonstrated in crude protein extracts from transgenic tobacco cell cultures using specific antibodies. Resveratrol, the product of stilbene synthase, was identified by HPLC and antisera raised against resveratrol.
一个编码芪合酶的花生（落花生）基因与一个嵌合卡那霉素抗性基因一起被转移。发现在烟草细胞（烟草）中用紫外线和激发子诱导后它能迅速表达。花生和转基因烟草悬浮培养物中芪合酶mRNA合成的比较研究揭示了相同的基因表达动力学。激发子诱导后30分钟和紫外线照射后10分钟可检测到芪合酶特异性mRNA。mRNA积累的最大值在诱导后2至8小时之间。诱导24小时后芪合酶mRNA积累停止。此外，在转基因烟草植株中，发现该基因在无菌根、茎和叶中可诱导。使用特异性抗体在转基因烟草细胞培养物的粗蛋白提取物中证实了芪合酶的存在。通过高效液相色谱法和针对白藜芦醇产生的抗血清鉴定了芪合酶的产物白藜芦醇。
