Rozman D, Komel R
Institute of Biochemistry, Medical Faculty, Ljubljana, Slovenia.
Curr Genet. 1992 Aug;22(2):123-7. doi: 10.1007/BF00351471.
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.
丝状真菌新月弯孢霉是一种已知的甾体11β-羟化酶，利用异源质粒pUT 720将其转化为博来霉素抗性。该质粒包含在构巢曲霉gpd和trpC表达信号控制下表达的Sh ble基因。博来霉素抗性菌落出现的频率为每微克DNA六个。所有菌落都是真正的转化体，未观察到“流产”生长。在所有测试的转化体中，质粒分子稳定地整合到宿主基因组中，并且其中一个质粒分子以位点特异性方式整合。转化体保留了甾体环羟化的能力，但羟基插入在15α位。
