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Expression of hepatitis B small surface antigen in Santalum album embryogenic cell suspension cultures

Shekhawat, U. K. S., Ganapathi, T. R., Srinivas, L.
Biologia plantarum 2010 v.54 no.4 pp. 720-724
Agrobacterium radiobacter, Santalum album, Western blotting, additives, culture media, enzyme-linked immunosorbent assay, genetically modified organisms, growing media, hepatitis B, hepatitis B antigens, kanamycin, reverse transcriptase polymerase chain reaction, surface antigens, trehalose
Embryogenic cell suspension cultures of Santalum album were transformed with Agrobacterium tumefaciens harboring pD35SHER plant expression vector having hepatitis B small surface antigen (HBsAg) with a C-terminal ER retention signal. The transformed colonies were selected on culture medium supplemented with kanamycin and subsequently the transgenic nature of these colonies was confirmed by PCR analysis. The expression of HBsAg was confirmed by RT-PCR analysis and Western blot analysis and the expression was quantified using monoclonal antibody-based ELISA. Cell suspension cultures were initiated from the colony with expression of 11.09 μg(HBsAg) g⁻¹(f.m.). To further increase the expression of HBsAg, transgenic S. album suspensions were cultured on media with various medium additives and cells growing in medium with 30 mM trehalose showed the expression of 19.95 μg(HBsAg) g⁻¹(f.m.).