ЭлБиб

303 / 592

PLANT GENETIC TRANSFORMATION, CELL SELECTION AND SOMATIC HYBRIDIZATION
USEOFPLASMIDSCARRYINGHEPATITISВ SURFACEANTIGENE FOR
CONSTRUCTIONOFVACCINEPRODUCINGTRANSGENICPLANTS
E.B.Rukavtsova, O.E.Zolova, Ya.I.Buryanov
Branch ofInstitute of Bioorganic Chemistry, PushchinoPushchino State University,
Pushchino, E-Mail:
Recently, transgenic plants producing foreign proteins (immunoglobulins,
viral surface antigenes), which play a role in human disease prophylaxis,
have been obtained. The aim of this work is the production of transgenic
tobacco plants expressing the Hepatitis В surface antigene
(HBsAg). DNA coding for HBsAg,which contains additional restriction
sites for cloning, was synthesized by polymerase chain reaction. The HBsAg
gene was cloned into pRT103 vector under control of CaMV 35S plant
promoter. Orientation of the insert was confirmed by restriction analysis of
this plasmid. The 35S-HBsAg-polyA cassette was subcloned into binary
vectors, pGA482 and pBiB-Hyg, which contain nptll gene for plant selection.
Agrobacterium tumefaciens LBA44Q4 (pAL4404) strain was transformed with
these plasmids. Agrobacterium tumefaciensstrain carrying pGA482::HBsAg
plasmid was used for inoculation of tobacco leaf discs. The shoot regenerants
were developed on selection medium with kanamycin and than were isolated
and cultured on the medium for rooting. The molecular biological and
biochemical analysis of these plants is in progress.
We propose that the use of Agrobacterium tumefaciensobtained strains
fortransformation of raspberry and strawberry plants can produce food
vaccines against Hepatitis B.
30 3
Научная электронная библиотека ЦНСХБ