Tazaki K, Yoshida K, Shinohara K, Koshiba T, Yamamoto N
Forestry and Forest Products Research Institute, Ibaraki, Japan.
FEBS Lett. 1995 Dec 11;377(1):54-8. doi: 10.1016/0014-5793(95)01254-0.
A cDNA encoding a bark lectin of Robinia pseudoacacia was introduced into tobacco plants. The expression of the lectin cDNA under control of the 35S promoter was confirmed by Western blot analysis and a hemagglutination assay of extracts of transgenic plants. Western blot analysis revealed that the subunit of the lectin from tobacco had a molecular mass of 29 kDa. The sequence of nine amino acids from the N-terminus of the lectin from transgenic tobacco plants was identical to that of the bark lectin from Robinia, indicating that the lectin had been processed correctly at its N-terminus in tobacco. The molecular mass of the purified native lectin produced by tobacco plants was estimated to be 112 kDa by gel filtration on a column of Superdex 200. It is suggested that the lectin subunits assembled to form tetramers in transgenic tobacco plants.
将编码刺槐树皮凝集素的cDNA导入烟草植株。通过蛋白质免疫印迹分析和转基因植物提取物的血细胞凝集试验,证实了凝集素cDNA在35S启动子控制下的表达。蛋白质免疫印迹分析显示,烟草凝集素的亚基分子量为29 kDa。转基因烟草植株凝集素N端九个氨基酸的序列与刺槐树皮凝集素的序列相同,表明该凝集素在烟草中N端加工正确。通过在Superdex 200柱上进行凝胶过滤,估计烟草植株产生的纯化天然凝集素分子量为112 kDa。提示该凝集素亚基在转基因烟草植株中组装形成四聚体。