Biolistic transient expression assay of Cm-eIF4E-Ved in C. zeyheri. (A) Schematic structure of MNSV and Cm-eIF4E constructs used in the transient expression assay. cDNAs were cloned into the binary vector pBIN61 between left (LB) and right (RB) borders of the Agrobacterium Ti plasmid. The 35S promoter and terminator are indicated as 35S-P and 35S-T, respectively. (B) RT-PCR detection of MNSV accumulation in bombarded leaves. pBMα5 (Mα5) and pB264 (264) constructs were bombarded separately and in combination with pB4E-PI (4E-PI) or pB4E-Ved (4E-Ved) into leaves of C. zeyheri. Two to three independent samples were included in the gel showed. Virus accumulation was assessed using RT-PCR two days post bombardment. C+ and C- indicate positive and negative controls of RT-PCR, respectively. C+ corresponds to leaves from susceptible melon bombarded with pBMα5 and pB264. In C-, RT-PCR was carried out with RNA from non-inoculated C. zeyheri leaves.
Nieto et al. BMC Plant Biology 2007 7:34 doi:10.1186/1471-2229-7-34