EXPRESSION OF KOREAN MISTLETOE LECTIN A-CHAIN GENE IN GENETICALLY MODIFIED NICOTIANA TABACUM

Abstract

Genetically modified (GM) tobacco (Nicotiana tabacum) harboring KMLA has been constructed to develop a plant expression system for the protein, KMLA. The 747 bp-long of the KMLA gene was ligated into BamHI/SacI- cut pCAMBIA3300 to obtain a recombinant plasmid, pCAMBIA3300-kmla. It was then transformed into Agrobacterium tumefaciens LBA4404 by liquid nitrogen method and the A. tumefaciens LBA4404/ pCAMBIA3300-kmla was inoculated into leaf discs of tobacco (20 days old). After co-cultivation, numerous calli were developed from transformed tobacco leaf explants which were cultured on MS basal medium containing kanamycin (Kan) (50 mg/ml), and then there were three GM tobacco plants generated from these calli. Transgenicity of the tobacco plants was confirmed by PCR screening and the KMLA gene expression was observed by Western blot.