DNA sequences required for anaerobic expression of the maize alcohol dehydrogenase 1 gene

Abstract

Expression of the maize alcohol dehydrogenase 1 (Adh1) gene is transcriptionally regulated under conditions of anaerobic stress. DNA sequences required for the expression of Adh1 have been identified by a functional analysis of in vitro constructed hybrid genes consisting of the Adh1 promoter fused to the chloramphenicol acetyltransferase coding region. A series of 5' deletions, 3' deletions, hybrid promoters, and linker scanning mutants of the Adh-CAT hybrid gene were introduced into maize protoplasts by electroporation and assayed for chloramphenicol acetyltransferase activity after incubation of the protoplasts under different oxygen tensions. The results indicate that a 40-base-pair DNA sequence within the Adh1 promoter is required for anaerobically regulated expression of the hybrid gene. Clustered point mutations in this sequence show that it is composed of two essential regions, each approximately 15 base pairs, separated by a 10-base-pair DNA sequence that does not appear to be important for anaerobic expression. Attachment of this 40-base-pair element to an unrelated promoter shows that this DNA sequence is both necessary and sufficient for induction of gene expression by low oxygen stress.