Sesame oleosin and prepro-2S albumin expressed as a fusion polypeptide in transgenic rice were split, processed and separately assembled into oil bodies and protein bodies

A recombinant polypeptide containing the precursor protein of a sesame storage protein, 2S albumin, fused to the C-terminus of a sesame oleosin was expressed in transgenic rice seeds under the control of a rice glutelin promoter. The recombinant polypeptide of 32 kDa, equivalent to the resultant molecular mass of sesame oleosin (15 kDa) and prepro-2S albumin (17 kDa), was detected in the endoplasmic reticulum fraction of maturing transgenic rice seeds, but not in the purified oil bodies or the soluble extract of transgenic seeds. However, sesame oleosin presumably fused with a 2 kDa C-terminal appendix originating from the signal sequence of prepro-2S albumin, was found in the purified oil bodies, and mature sesame 2S albumin apparently processed into two subunits (9 and 4 kDa) linked by disulfide bonds was detected in extracts of transgenic seeds. Immunogold labeling revealed that the sesame oleosin and 2S albumin were separately located in oil bodies and protein bodies of embryo cells of transgenic rice seeds. While sesame 2S albumin was also detected in protein bodies of endosperm cells of transgenic seeds, the co-expressed sesame oleosin, probably degraded due to the lack of oil bodies in this tissue, and was not detected. The results provide a new technique for introducing two recombinant polypeptides separately into rice oil bodies and protein bodies from one expression construct.