Abstract
BBM (baby boom) is an important embryogenesis transcription factor in plants, which is valuable in molecular research and could improve the transformation efficiency in recalcitrant species. However, in time and dose aspect, which stage and how much BBM transcription factor is necessary for stimulating embryogenesis is still largely unknown in Brassicas. GR system and dexamethasone are important to timing and quantitative analysis of transcription factors. In this study, BP and LR reaction were applied to build czn3999-35s: Omega-BBM-GR construct, which was transformed into <i>E. coli</i> DH5 α and <i>Agrobacterium </i>C58 competent cells by electric shock transformation respectively. The correct strain verified by PCR digestion reaction was kept in glycerol and stored at -80°C. <i>Brassica napus</i> 12075 cotyledon petioles were used as recipient, and eventually 22 transgenic plantlets were obtained from agrobacterium mediated bulk-way transformation. We describe Bulk-Way transformation protocol in detail, and provides valuable materials for analyzing dose and time effect of BBM in Brassicas.
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