biologia plantarum

International journal on Plant Life established by Bohumil Nìmec in 1959

Biologia plantarum 54:720-724, 2010 | DOI: 10.1007/s10535-010-0128-6

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

U. K. S. Shekhawat1, T. R. Ganapathi1,*, L. Srinivas1
1 Plant Cell Culture Technology Section, Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre, Trombay, Mumbai, India

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.).

Keywords: Agrobacterium tumefaciens; C-terminal ER retention signal; genetic transformation; medium additives
Subjects: Agrobacterium tumefaciens; calcium; gibberellic acid, gibberellin; in vitro culture, cell suspension; mannitol; nitrogen supply; polymerase chain reaction (PCR); Santalum album; transgenic plants; trehalose; Western blot analysis

Received: January 16, 2009; Accepted: August 15, 2009; Published: December 1, 2010  Show citation

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Shekhawat, U.K.S., Ganapathi, T.R., & Srinivas, L. (2010). Expression of hepatitis B small surface antigen in Santalum album embryogenic cell suspension cultures. Biologia plantarum, 54(4), 720-724. doi: 10.1007/s10535-010-0128-6
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