EVALUATION OF T3 GENERATION TRANSGENIC TOBACCO WITH HEPATITIS B GENE FOR GROWTH, YIELD AND MOLECULAR CHARACTERS
Loading...
Date
2010-07-21
Authors
Journal Title
Journal ISSN
Volume Title
Publisher
University of Agricultural Sciences GKVK, Bangalore
Abstract
Hepatitis B virus (HBV) is a major cause of acute and chronic hepatitis.
The current HBV vaccine is a biotechnology product that falls in the category of
subunit vaccines and is made from yeast cells grown by fermentation. In recent
years, a novel production system of vaccines-edible vaccines has been developed.
Edible vaccines can serve multiple immunization priorities and offer advantages,
including simplicity of use, lesser expense, enhanced immune responses at
mucosal sites, and stimulation of humoral immunity. The plant-based production
of vaccines provides new opportunities to develop oral vaccines for hepatitis B.
Different host systems were employed to produce hepatitis B. In addition, a plant
based HBsAg expression system makes possible the testing of an oral
immunization strategy by simply feeding the plant samples. The present
investigation lays emphasis on study of stability of the recombinant protein
expressed in tobacco plants. Restriction digestion analysis of the gene construct
pHB118 with restriction enzymes EcoRI and BamHI yielded two separate bands
of 9.7kb and 3.6kb size. The presences of HBsAg gene in T3 transgenic plants
were confirmed by PCR analysis. The transgenic tobacco plants showed the
presence of 900 bp band in the PCR analysis. The crude protein obtained from the
transformed tobacco plants were tested by SDS-PAGE for the presence of 24 kDa
protein, and ELISA confirmed the antigen specificity and immunogenic nature of
the Hepatitis B surface antigen. The T3 generation seeds obtained from the
transgenic tobacco plants were tested for the germination in presence of
kanamycin. It was observed that the segregation ratio was 3:1 indicating
Mendelian inheritance. The growth parameters of T3 generation transgenic and
control tobacco plants showed slight variations in growth between transgenic and
control plants.
Description
Keywords
null