Genetic introduction of thyroid hormone transporter in Bood-Brain-Barrier

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We have established adenovirus-mediated gene transfer system into cells.The coding region of LST-1 and LST-2, both of which have isolated by us have introduced into adenovirus vector, packaged, and transfected into HEK293 cell with adenovirus genomic DNA.The cells that produces homologous-recombinant shrieked and made plaque formation. After producing large scale infected medium, the recombinant adenovirus was purified by CsCl ultracentrifuge.The infection titer of the purified LST-1 and LST-2 adenoviruses were measured and the IC 50 of each virus was 1.26 and 1.48 X 10^<10> pfu, respectively.When recombinant virus was applied to cultured cell line, the LST-1 and LST-2 protein was strongly expressed within 24 hr after application. The size of the band was also confirmed by Western blot analysis. The LST-1 and LST-2 expressing mammalian cell transports various organic anions as substrates and the apparent Km value of tauro-cholate uptake is 24 micro M and 32 micro M, respectively and both of which is a comparative value of the native cells reported.The establishment of the adenovirus-mediated gene transfer system wiil lead to the various applied sysem for screening of the brain-, liver and kidney oriented drug delivery and therapeutic tools.