Expression of cDNA for a bark lectin of Robinia in transgenic tobacco plants

FEBS Letters

Yamamoto, Naoki¹  Yoshida, Kazumasa²  Koshiba, Tomokazu³  Shinohara, Kenji²  Tazaki, Kiyoshi² 

[1] National Institute of Agrobiological Resources, Tsukuba, Ibaraki 305, Japan;Forestry and Forest Products Research Institute, Kukizaki, Ibaraki 305, Japan;Department of Biology, Tokyo Metropolitan University, Hachioji, Tokyo 192-03, Japan

关键词: Robinia pseudoacacia;    Bark lectin;    Transgenic tobacco;    Hemagglutinating activity;    CaMV;    cauliflower mosaic virus;    CBB;    Coomassie brilliant blue;    FITC;    fluorescein isothiocyanate;    GUS;    β-glucuronidase;    PBS;    phosphate-buffered saline;    PCR;    polymerase chain reaction;   

DOI  :  10.1016/0014-5793(95)01254-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown