[Show abstract][Hide abstract] ABSTRACT: The development of modern gene technologies allows for the expression of recombinant proteins in non-native hosts. Diversity in translational and post-translational modification pathways between species could potentially lead to discrete changes in the molecular architecture of the expressed protein and subsequent cellular function and antigenicity. Here, we show that transgenic expression of a plant protein (alpha-amylase inhibitor-1 from the common bean (Phaseolus vulgaris L. cv. Tendergreen)) in a non-native host (transgenic pea (Pisum sativum L.)) led to the synthesis of a structurally modified form of this inhibitor. Employing models of inflammation, we demonstrated in mice that consumption of the modified alphaAI and not the native form predisposed to antigen-specific CD4+ Th2-type inflammation. Furthermore, consumption of the modified alphaAI concurrently with other heterogeneous proteins promoted immunological cross priming, which then elicited specific immunoreactivity of these proteins. Thus, transgenic expression of non-native proteins in plants may lead to the synthesis of structural variants possessing altered immunogenicity.
Journal of Agricultural and Food Chemistry 12/2005; 53(23):9023-30. · 3.11 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: We describe a robust and reproducible Agrobacterium-mediated chickpea transformation method based on kanamycin selection, and its use to introduce the bean AI1 gene into a desi type of chickpea. Bean AI1 was specifically expressed in the seeds, accumulated up to 4.2% of seed protein and was processed to low molecular weight polypeptides as occurs in bean seeds. The transgenic protein was active as an inhibitor of porcine -amylase in vitro. Transgenic chickpeas containing -AI1 strongly inhibited the development of Callosobruchus maculatus and C. chinensis (Col. : Bruchidae) in insect bioassays.
[Show abstract][Hide abstract] ABSTRACT: Bruchid larvae cause major losses of grain legume crops through- out the world. Some bruchid species, such as the cowpea weevil and the azuki bean weevil, are pests that damage stored seeds. Others, such as the pea weevil (Bruchus pisorum), attack the crop growing in the field. We transferred the cDNA encoding the a-amylase inhibitor (a-AI) found in the seeds of the common bean (Phaseolus vulgaris) into pea (Pisum sativum) using Agrobacferium-mediated transformation. Expression was driven by the promoter of phytohe- magglutinin, another bean seed protein. The a-amylase inhibitor gene was stably expressed in the transgenic pea seeds at least to the T, seed generation, and a-AI accumulated in the seeds up to 3% of soluble protein. This level is somewhat higher than that normally found in beans, which contain 1 to 2% a-AI. In the 1, seed generation the development of pea weevil larvae was blocked at an early stage. Seed damage was minimal and seed yield was not significantly reduced in the transgenic plants. These results confirm the feasibility of protecting other grain legumes such as lentils, mungbean, groundnuts, and chickpeas against a variety of bruchids using the same approach. Although a-AI also inhibits human a-amy- lase, cooked peas should not have a negative impact on human energy metabolism.