Cloning and expression of Kluyveromyces fragilis LAC4 gene

The genomic library of Kluyveromyces fragilis was constructed in E. coli TG1, and 5 beta-galactosidase gene (LAC4) clones have been obtained from the library by complementation of the Kluyveromyces lactis lac4-8 mutation. The studies on the structure and the function of the LAC4 gene revealed that (i) the gene can also complement E. coli lacZ mutation; (ii) the physical map of the K. fragilis LAC4 gene was very similar to that of K. lactis; (iii) the beta-galactosidase levels expressed by the clone strains were much higher than that expressed by the original strain; (iv) the variation of the beta-galactosidase level of different clone strains induced by lactose or galactose was related to the retained degree of the 5' flanking region of LAC4 gene, suggesting that there might be a lactose specific transcription activating element in the region.