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Pea lectin is correctly processed,stable and active in leaves of transgenic potato plants
Authors:Glyn A Edwards  Andrew Hepher  Stephen P Clerk  Donald Boulter
Institution:(1) Department of Biological Science, Durham University, DH1 3LE Durham, Co. Durham, UK;(2) Shell Research Ltd., Sittingbourne Research Centre, ME9 8AG Sittingbourne, Kent, UK;(3) Present address: Shell Research Ltd., Sittingbourne Research Centre, ME9 8AG Sittingbourne, Kent, UK
Abstract:A gene encoding the preproprotein of the pea (Pisum sativum) lectin was expressed in transgenic potato plants using a cauliflower mosaic virus (CaMV) 35S promoter or a tobacco ribulose bisphosphate carboxylase small subunit (ssRubisco) promoter. Presence of the pea lectin to levels greater than 1% of total soluble leaf protein was detected by radioimmunoassay (RIA). The pattern of expression derived from the two promoters was established using both RIA and a squash-blot immunolocalisation technique. Western blotting demonstrated that the preproprotein was correctly processed, generating agr and beta subunits that assembled to give an isolectin form observed in pea seeds and roots. It was also found that the haemagglutination activity and specificity of pea lectin synthesised in transgenic potato leaves was comparable to purified lectin from pea cotyledons.
Keywords:Agrobacterium  haemagglutination  lectin (Pisum)  protein processing  transgenic potato
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