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Heterologous Expression of ABC Transporter in Catharanthus Roseus (L.) G. Don
 Pomahačová, Barbora ; Dušková, Jiřina (advisor) ; Tůmová, Lenka (referee) ; Grančai, Daniel (referee)
In this thesis, the regulation of transport is investigated as a limiting factor in biosynthesis of plant secondary metabolites. CjMDR1 in Catharanthus roseus (L.) G. Don. has been selected as a model system of active transporters. CjMDR1 is a member of ABC (ATP-Binding Cassette) protein superfamily. Especially, in previous reports it was found to transport preferably berberine in its producing plant, Coptis japonica. In order to look into the role of the ABC transporter berberine was exogenously fed to Catharanthus roseus cell line and evaluate the transport of the compound. Based on a series of experiments in this study it can be concluded as follows. 1. Transport of berberine in wild Catharanthus roseus was not clearly associated with ATP-dependency. Using diverse inhibitors it was proved that berberine was not an exclusive substrate of native ABC transporters in Catharanthus. Alternatively berberine might be transported via H+-antiport or efflux transporters from MATE (multidrug and toxic compound extrusion) group which was previously found in other heterologous plants (Yazaki, 2005). In reality in Catharanthus some effect of pH on plasmatic membrane on berberine transport has been observed. However, the exact mechanism of berberine transport on vacuolar and cell level was not fully uncovered...
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