Hemagglutination activity assay of the rice Gal_lectin member LOC_Os03g06940. (A) Domain structures of the positive control (up) and the Gal-lectin member detected by Pfam database. (B) In vitro expression of the recombinant proteins as shown by rectangles. M, molecular marker; 1, expressed GST tag in the vector pGEX-6p as a negative control; 2, recombinant protein from the Gal_lectin domain and its flanking region; 3, from the Glyco_hydro_35 domain; 4, exactly from the Gal_lectin domain; 5, from the Jacalin domain as a positive control. (C) and (D) No agglutination of the erythrocytes was observed when E. coli extracts from the empty vector (lane1 in B) or from the recombinant construct containing the Glyco_hydro_35 domain (lane 3 in B) were used for the activity assay. (E) and (F) Agglutination of erythrocytes by the E. coli extract from positive control (E) or from the recombinant construct containing the Gal_lectin domain (F). Recombinant proteins from both lane 2 and 4 show the agglutination activity and only the latter is shown in (F).