In vitro and in vivo study of the activity of secreted alkaline phosphatase mRNA ribozyme gene
Zakharchuk, A.N.; Doronin, K.K.; Karpov, V.A.; Krugliak, V.A.; Naroditskiĭ, B.S.
Molekuliarnaia Genetika Mikrobiologiia i Virusologiia 6: 8-12
1994
ISSN/ISBN: 0208-0613 PMID: 7739597 Document Number: 426045
A ribozyme was constructed of the catalytic domain of tobacco ringspot virus satellite RNA and flanking sequences complementary to the target, secreted alkaline phosphatase (SEAP) mRNA. The ribozyme specifically cleaved the substrate in vitro; compared with that constant temperature. The relationship between specific endoribonuclease activity of the ribozyme and Mg2+ concentration was shown. The ribozyme was active in 293 cell line which was cotransfected with plasmids carrying the SEAP gene under control of RSV LTR promoter and the ribozyme gene under early HCMV promoter: SEAP activity reduced by half.