Functional examination of the generated GatewayRdestination vectors. Full-length cDNA sequence of BAK1 was PCR amplified from Arabidopsis Col-0 plants and introduced into two destination vectors, pB35GWG and pB35GWF, by in vitro DNA recombination mediated by BP clonase and LR clonase. Expression constructs harboring BAK1 cDNA were transformed into the bri1-5 mutant, a weak allele of bri1. Overexpression of BAK1 using the vectors can suppress the bri1-5 mutant phenotype, indicating that the vectors are functional. Expression of BAK1 was confirmed by Western hybridization in transgenic plants. Anti-FLAG and anti-GFP antibodies were mixed to detect the signals on one membrane.
Gou et al. BMC Genomics 2010 11:19 doi:10.1186/1471-2164-11-19