A One-Time Inducible Transposon to Create Knockout Mutants in Rice

Use of a transposon is an efficient tagging tool for exploring the function of the gene it inserts into or is adjacent to. A few modifications have been applied to the native Ac transposon to allow it to transpose efficiently or spontaneously and stop quickly thereafter. Furthermore, locating the transposon between a constitutive plant promoter and a reporter gene, such as the firefly luciferase gene, allows for nondestructively detecting excision events in vivo . This chapter describes a detailed protocol for one-time inducible transposon tagging of rice cells and their subsequent screening and regeneration into mutant lines.