The gene from the dimorphic fungus was cloned by functional complementation of the arginine auxotrophy present in strain EL2 (Arg) using a gene library constructed in the double autonomously replicating sequence vector pRM1. Sequence analysis revealed a putative 857 amino acid polypeptide (95 kDa) which showed high homology (63% protein identity) to the 6 gene. Similarly to the gene, the gene is responsible for both the acetylglutamate kinase and acetylglutamyl-phosphate reductase activities, the second and third steps of arginine biosynthesis at the mitochondria. The gene complemented the mutation present in (strain D160-4D) on a yeast episomal plasmid using its own regulatory signals. A set of non-integrative high-efficiency plasmid vectors based on this gene marker was constructed and a null ,6δ strain was obtained using the common -blaster strategy. In addition, we generated an ,6δ null mutant in a single transformation event, thus improving the basic strategy for generating gene deletions in


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