FUNCTIONAL-ANALYSIS OF LINKER INSERTIONS AND POINT MUTATIONS IN THE ALPHA-AMY2 54 GA-REGULATED PROMOTER/

Functional analysis of a gibberellin-regulated wheat a-amylase promote r, alpha-Amy2/54, has indicated that three regions were essential for expression. By studying the ability of mutant promoters, containing a randomly inserted 22 bp excision linker, to direct expression in oat a leurone protoplasts we have refined the positions and extents of these three cia elements and also demonstrated the presence of two addition al elements. By converting the linker insertions to either single base point mutations or deletions using the class IIS restriction endonucl ease Bsm I we have shown that nucleotides -119 and -109 within the GAR E (-121)GTAACAGAGTCTGG(-108) and nucleotide -152 within the proposed e lement (-156)GATTGACTTGACC(-144) are essential for high level expressi on from this promoter.