Cloning of Soybean 24 kDa Oleosin Gene and Its Transient Expression asa Carrier for Foreign Protein

The genomic DNA sequence encoding soybean 24 kDa oleosin and its promoter were cloned and analyzed for investigation of the potentials of the oleosin acted as a carrier for production of recombinant proteins in plant. The -300 box, GA-rich, G-box, SEF-3, SEF-4, RY box, ABA box, CAn and TATA box were found in the upstream region of the soybean oleosin gene, which shows the functional oleosin promoter available. Homology comparison reveals that the soybean 24 kDa oleosin shares the highest identity with the soybean oleosin isoform A (U09118, GenBank), reaching to 98.4% in nucleotide. A soybean oleosinhirudin fusion gene driven by the oleosin promoter was constructed and inserted into plant binary expression vector. The intact tobacco plantlets were transformed by means of vacuum infiltration approach, with the Agrobacterium tumefaciens harboring the above vector. The transient correct expression of oleosin-hirudin fusion gene was identified by SDS/PAGE, western blotting and enterokinase treatment.