Immunoblotting Analysis of nsLTPl in Cereal Grains with Antiserum Raised against Recombinant Rice nsLTP1

摘要

Nonspecific lipid transfer protein 1 (nsLTPl) is widely distributed in plants and has been recognized as a widely cross-reacting plant pan-allergen. In the present study, a recombinant protein of rice nsLTPl was prepared with an Escherichia coli expression system, and polyclonal antiserum was raised by immunizing rabbits with this recombinant nsLTPl. The antiserum specifically immunoreacted with rice nsLTPl and homologous nsLTPl proteins of other kinds of cereal grains. Moreover, the antiserum was applied to compare nsLTPl content among rice cultivars and among rice fractions separated by grain-milling. Variation of nsLTPl contents were observed among rice cultivars, and rice nsLTPl was proved to be mainly located in the outer layer of rice grains, providing evidence for decreasing nsLTPl content by grain-milling.