Transformation of Cochliobolus lunatus with pUT 720 changes the steroid hydroxylating ability of the fungus

Abstract

The filamentous fungus Cochliobolus lunatus, a known 11 beta-hydroxylator of steroids, was transformed to bleomycin resistance using the heterologous plasmid pUT 720. This plasmid contains the Sh ble gene expressed under the control of the Aspergillus nidulans gpd and trpC expression signals. The bleomycin-resistant colonies appeared with a frequency of six per microgram of DNA. All colonies were real transformants and no "abortive" growth was observed. In all transformants tested the plasmid molecules became stably integrated into the genome of the host, and one of the plasmid molecules integrated in a site-specific manner. Transformants retained the ability to hydroxylate the steroid ring, but the hydroxy group was inserted at the 15 alpha position.