Fig. 5
Application analysis of antisera by western blotting. a, b Expression comparisons of CABYV and MABYV in N. benthamiana leaves inoculated with full-length infectious clones by western blotting and RT-PCR. Marker: pageruler prestained protein ladder; Mock: inoculated pCass-RZ empty vector; Lanes 1, 2, 3: leaf samples expressing CABYV or MABYV infectious clone; “+”: proteins extracted from N. benthamiana leaves transiently expressing MPCABYV or MPMABYV. Rubisco stained with coomassie brilliant blue is used as a loading control (middle panel). The lower panels are RT-PCR data from the same samples. c Simulation detection of cucumber samples infected with CABYV, MABYV or SABYV. Marker: pageruler prestained protein ladder; Mock: proteins extracted from healthy cucumber leaves; EV: N. benthamiana leaves inoculated with pMDC32 empty vector; “+”: protein extracted from N. benthamiana leaves transiently expressing the three MP genes; The other lanes show different dilutions of proteins extracted from N. benthamiana leaves transiently expressing MPs (1:16, 1:32, 1:64, 1:128, 1:256 and 1:512) and mixed with proteins extracted from healthy cucumber leaves. The three antisera were used at 1:10000