[Prokaryotic expression of wheat <i>TaABI5-D3</i> gene and polyclonal antibody preparation].

Abscisic acid insensitive 5 (ABI5) is a basic leucine zipper transcription factor regulating ABA-mediated seed germination, and <i>TaABI5</i> is closely related to the pre-harvest sprouting in wheat. Studies have shown that <i>TaABI5-D3</i>, one of multiple copies of <i>TaABI5</i> gene, encodes the intact TaABI5 protein. In this study, we constructed the prokaryotic expression vector pET-28a-<i>TaABI5-D3</i> for expression of <i>TaABI5-D3</i> in <i>Escherichia coli</i> and obtained the purified recombinant protein His-TaABI5-D3. This protein existed in the form of inclusion bodies, with the best expression induced by incubation with 0.6 mmol/L IPTG at 16 ℃, 150 r/min overnight (for 12 h). Subsequently, the purified His-TaABI5-D3 was injected into Balb/C mice for the preparation of polyclonal antibodies. Western blotting analysis indicated that the polyclonal antibody had relatively high specificity, laying foundations for clarification of the function of TaABI5 protein in wheat.