Abstract

Ribosome-inactivating proteins (RIPs) are toxic proteins synthesized by many plants and some bacteria, that specifically depurinate the 28S RNA and thus interrupt protein translation. RIPs hold broad interest because of their potential use as plant defense factors against pathogens and cancer cells. The most promising way to exploit plant RIPs as weapons against cancer cells is either by designing molecules in which the toxic domains of RIPs are linked to selective tumor targeting domains or directly delivered as suicide genes for cancer gene therapy. In this research the sequence of rice RIP gene was synthesized in PUC57 plasmid. The gene was cloned in pPICZαA vector and expressed in Pichia pastoris KM71H (mut^s) strain. The expected protein which had an apparent molecular mass of 34 kDa was detected by SDS-PAGE analysis and confirmed by Western blot.

Key words: Ribosome-inactivating proteins, Pichia pastoris, N-glycosidases.

Abbreviation

Abbreviations: SDS-PAGE: Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis; RIP: Ribosome-inactivating proteins; EGFR: epidermal growth factor receptor; IT: Immunotoxin