Designing and construction of Gamma Interferon- Oleosin fusion and transformation in Brassica napus

The oleosin fusion protein provide a unique route for the large-scale production of recombinant proteins in plants, as well as an efficient process for purification of desired polypeptide. By fuseing oleosin to desired gene and using seed specific promoter,targeting of recombinant protein to seed oil bodies will be possible. In this study, we used plant oleosin as a carrier for the production of the human gamma interferon protein in Brassica napus seeds. At the first step, oleosin gene was isolated from Brassica napus genome and cloned in pBI121 plant binary vector. Human gamma interferon gene was cloned downstream of oleosin and proteolytic cleavage site was set between them. The interferonoleosin fusion gene was under the control of seed specific promoter (Napin). Cotyledon explant and A. tumefaciens (LBA4404) were used for plant transformation. The transformed shoots were screened on kanamycin contained media. The presence and expression of the transgene was confirmed in the transformants by PCR and RT-PCR.