Candida glabrata shuttle vectors suitable for translational fusions to lacZ and use of beta-galactosidase as a reporter of gene expression

The functionality of P-galactosidase encoded by the E. coil lacZ gene as a reporter of gene expression in C. glabrata was investigated. C. glabrata/E. coli shuttle vectors were constructed, containing both a C. glabrata CEN-A RS cassette, to allow regular segregation and episomal replication of the p lasmids, and the lacZ coding sequence of E, coil. The functionality of P-ga lactosidase in C. glabrata was verified by inserting the promoter and the 5 ' coding region of the HIS3 gene from C. glabrata directionally upstream of the lacZ gene. By fusing the promoter of the copper-controlled MTII gene t o the lacZ reporter, we showed that P-galactosidase activity can be differe ntially induced in C. glabrata. P-galactosidase reporter activities were de tected qualitatively by an indirect filter assay and quantitatively from pe rmeabilized cells. (C) 2000 Elsevier Science B.V. All rights reserved.