Expressing of the human interferon gamma gene in the Tobacco chloroplast
Message:
Abstract:
Interferon gamma (IFN-g) is one of the most important proteins used in diagnostic and therapeutic especially in virus disease therapy and regulation of immune system actions. There are several ways for production of interferon gamma protein as an active ingredient of many medicines and one of these ways is using plants and molecular farming. In latest years chloroplast transformation due to having many advantages such as ability to accumulate large amounts of foreign protein in chloroplast, being free of human pathogens and expression of several genes simultaneously has been considered. The purpose of this research was to check feasibility of interferon gamma production using chloroplast transformation in a dicistronic operon. Interferon gamma gene was cloned in pKCZ vector. Then gold particles coated with this vector were bombarded on tobacco leaves by gene gun. The pieces of leaves were transferred to shoot regeneration medium containing spectinomycin. Thereafter the regenerated shoots were transferred to MS medium for rooting. Presence of interferon gamma gene and it’s expression in regenerated plants was confirmed by PCR and RT-PCR. Additionally gene expression in protein level was confirmed by SDS-PAGE and dot blot. Finally four regeneration rounds were performed to obtain homoplasmy.
Language:
Persian
Published:
Journal of Genetics, Volume:8 Issue: 4, 2014
Pages:
333 to 340
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