Analysis of the gene for the elongation factor 1 alpha from the zygomycete Absidia glauca. Use of the promoter region for constructions of transformation vectors

Abstract

The complete genomic DNA sequence was determined for one of the gene for the elongation factor 1 alpha (TEF), isolated from the zygomycete Absidia glauca. Sequence comparison with TEF genes from other fungi show the highest similarity to TEF-genes of the closely related zygomycete Mucor racemosus (Sundstrom et al. 1987). Southern-blot analysis of genomic DNA from A. glauca with the TEF gene reveals six chromosomal copies in the genome. In transformation experiments of A. glauca, vector constructions were used which allow targeting of one of the TEF loci. Several transformants of A. glauca were analyzed at the DNA level. In most cases, rearranged forms of autonomously replicated plasmids could be found in these isolates. However, some transformants show a different restriction pattern of the TEF loci if compared with the parental strains. From Southern-blot data it could be concluded that in one case the rearrangement lies downstream of one TEF locus. In a second case genetic parts following the 3'-end of the TEF gene are moved towards the 5'-end of the gene.