Expression of human growth hormone in transgenic rice cell suspension culture

Abstract

Human growth hormone (hGH), a pituitary-derived polypeptide, evidences a wide range of biological functions, including protein synthesis, cell proliferation, and metabolism. A synthetic hGH gene (shGH) has been synthesized on the basis of plant-optimized codon usage via an overlap PCR strategy and located in a plant expression vector under the control of the rice amylase 3D (Ramy3D) promoter, which is induced by sugar starvation. The plant expression vector was introduced into rice calli (Oryza sativa L. cv. Donjin) via particle bombardment transformation methods. The integration of the shGH gene into the chromosome of the transgenic rice callus was verified via genomic DNA PCR amplification and shGH expression in transgenic rice suspension cells was confirmed via Northern blot analysis. The shGH protein was detected in the transgenic rice cell suspension culture medium following induction with sugar starvation, using Western blot analysis. The quantity of shGH that accumulated in the transgenic rice cell suspension medium was 57 mg/l. The shGH accumulated in the transgenic rice cell suspension culture medium evidenced a biological activity similar to that of Escherichia coli-derived recombinant hGH. These results indicate that the shGH was generated and accumulated in the transgenic rice cell suspension culture medium, and manifested biological activity.

High-Level Transient Expression of ER-Targeted Human Interleukin 6 in Nicotiana benthamiana

“…These induce the formation of protein bodies (PB's) as storage organelles in leave tissue, which are devoid of proteolytic activity [67]. The impact of cellular proteases can also be reduced by coexpressing a protease inhibitor, or the development of specialized plant lines lacking protease activity [68]–[70].…”

Expression of functional recombinant human growth hormone in transgenic soybean seeds

“…Expression of recombinant hGH protein in plants has been reported in transgenic tobacco (Nicotiana tabaccum) seeds (Leite et al 2000), chloroplasts (Staub et al 2000), the apoplast of leaves of Nicotiana benthamiana (Gils et al 2005), rice cell suspensions (Kim et al 2008), tobacco cell suspension, corn seeds and soybean [Glycine max (L.) Merrill] seeds and leaves (Russell et al 2005). In a recent study by Rabindran et al (2009), a plant virusbased expression vector was utilised to transiently express a biologically active hGH in N. benthamiana plants.…”

Molecular Farming in Plants

“…c. Plant cell suspension culture: This method involves the removal of cell walls and gene transfer to the obtained protoplasts and suspension culture. The purification system and its downstream processing are cheaper and easier [17]. In addition, the use of suspension culture can decrease heterogeneity in proteins and sugar (N-glycans) regarding the uniformity of the type and size of cells [5].…”