[Production of marker-free plants expressing the gene of the hepatitis B virus surface antigen].
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.
Rukavtsova
E B
EB
Gaiazova
A R
AR
Chebotareva
E N
EN
Bur'ianova
Ia I
IaI
D005819:Genetic Markers; D006514:Hepatitis B Surface Antigens / Q000096:biosynthesis* / Q000235:genetics; D006515:Hepatitis B virus; D018551:Solanum lycopersicum / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism*
D005819:Genetic Markers; D006514:Hepatitis B Surface Antigens