Production of marker-free plants expressing the gene of the hepatitis B virus surface antigen |
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Authors: | E B Rukavtsova A R Gayazova E N Chebotareva Ya I Buryanov |
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Institution: | (1) Department of Horticulture, Agricultural College, Shihezi University, Shihezi, 832003, People’s Republic of China |
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Abstract: | The pBM plasmid, carrying the gene of hepatitis B virus surface antigen (HBsAg) and free of any selection markers of antibiotic or herbicide resistance, was constructed for genetic transformation of plants.
A method for screening transformed plant seedlings on nonselective media was developed. Enzyme immunoassay was used for selecting
transgenic plants with HBsAg gene among the produced regenerants; this method provides for a high sensitivity detection of HBsAg in plant extracts. Tobacco
and tomato transgenic lines synthesizing this antigen at a level of 0.01–0.05% of the total soluble protein were obtained.
The achieved level of HBsAg synthesis is sufficient for preclinical trials of the produced plants as a new generation safe
edible vaccine. The developed method for selecting transformants can be used for producing safe plants free of selection markers. |
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