As. Pac. J. Mol. Biol. & Biotech., Apr 2013 Vol. 2, -
Characterization of T2 Tobacco Plants expressing the Hepatitis B gene for Molecular and Morphological Characters
Asha Velayudhan Nair1*, Yogendra Kalenahalli Narasimhamurthy2, Paramanahally Hanumanthegowda Ramanjini Gowda3
1School of Biochemistry, University of Bristol, United Kingdom.
2Plant Science Department, McGill University, Montreal, Quebec, Canada.
3Department of Biotechnology, University of Agricultural Sciences, Bangalore, India.
* Author for correspondence: Asha Velayudhan Nair
School of Biochemistry, Medical Sciences Building, University of Bristol, BS8 1TD, United Kingdom.
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Abstract.
Hepatitis B virus (HBV) infection remains a worldwide health problem and immunization is the only means to prevent the occurrence of this disease. As the available vaccines are very expensive, there is still need for a less expensive vaccine source especially in most of the developing countries. Plants provide great advantages over other expression systems for the production of recombinant proteins effectively at low cost. In this paper we have shown the stable expression of the hepatitis B surface antigen (HBsAg) in transgenic tobacco plants into the second generation; the current primary focus in plant transformation is the stability of transgene expression. The presence of the HBsAg gene in putative transgenics of the T2 generation of tobacco plants was confirmed by PCR analysis that produced a 900 bp amplification product. Transgenic and untransformed (control) tobacco plants were evaluated for the presence and expression of the hepatitis B surface antigen gene by SDS-PAGE, protein dot blot and ELISA. The T2 generation seeds obtained from the transgenic tobacco plants were tested for germination rate in the presence of kanamycin and were observed to have a segregation ratio of 3:1, indicating Mendelian inheritance. Observations of growth and yield parameters of T2 generation tobacco plants transformed with the HBsAg gene were recorded at regular intervals. It was found that the presence of the foreign gene did not inhibit the growth of the transgenic plants; transgenics showed similar growth rate compared to that of the control plants.