5.

Cma ST8Sia IV expression in a heterologous plant system. (A) The total protein (TP) extracts from leaves infiltrated with Cma ST8Sia IV constructs with two distinct mammalian Sia pathways, N-Sia (upper) and O-Sia pathway (lower), were monitored by WB using mAb735. In addition, aliquots of PNGase F- or endoN-treated samples were analyzed. TP from leaves infiltrated without Cma ST8Sia IV was used as a negative control. (B) Polysialylated proteins extracted from leaves were enriched by ion exchange resin and inactive endoN affinity precipitation. Chain length analysis by anion exchange HPLC was performed for Cma ST8Sia IV with the N-Sia pathway and Cma ST8Sia IV with the O-Sia pathway.