Relative strengths of promoters from Cochliobolus heterostrophus

Abstract

GPD1 (encoding glyceraldehyde-3-phosphate dehydrogenase) is a constitutively expressed gene in Cochliobolus heterostrophus that produces a single transcript. The steady state level of GPD1 mRNA is 14-fold greater than that of the constitutively-expressed TRP1 gene (encoding a tryptophan biosynthesis enzyme) indicating that GPD1 has a stronger promoter and/or a more stable mRNA. A set of lacZ translational fusion vectors was constructed to compare the gene expression signals of GPD1, TRP1 and PRO1 (a C. heterostrophus genomic fragment selected for promoter activity) in C. heterostrophus as single copies at the same site in the chromosome. Under conditions that repressed endogenous beta-galactosidase expression, beta-galactosidase activity in transformants was constitutive and required the GPD1, TRP1 or PRO1 expression signals. In-frame GPD1::lacZ activities were 6-fold greater than in-frame TRP1::lacZ and PRO1::lacZ activities, indicating that GPD1 has more efficient expression signals.