Figure 6

Molecular analysis of transgenic lines under evaluation in confined field trial.

(a) RT-PCR of 10 transgenic plantain lines (peptide only and dual stacked genes lines) using primers specific to peptide and 25S ribosomal gene. The figure shows cropped gels and both gels were run under the same experimental conditions. Amplified RT-PCR product designations were shown on the right and products sizes were shown on the left. M - molecular weight marker, NTC - non-transgenic control; (b) Relative transcript levels of the peptide in 10 transgenic lines (peptide only and dual stacked genes lines). Expression of peptide was normalized with banana 25S ribosomal gene (internal control) and the non-transgenic plant served as a calibrator. Relative expression was determined from replicate measurements in two independent biological replicates and three technical replicates. Values are means ± SEM. (c) Detection of cystatin expression by western blot for seven transgenic plantain lines (cystatin lines and dual stacked genes line) using protein extracted from roots pooled of four replicate plants in each block of the contained field trial. The molecular weight of the transgenic maize cystatin is indicated (). Non-transformed Gonja manjaya control (NTC) protein extract was spiked with the indicated quantity of 6 × His-tagged cystatin for positive controls. The figure shows cropped blots and gels were run under the same experimental conditions.