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Heterologous transformation of Mucor circinelloides with the Phycomyces blakesleeanus leu1 gene
E A Iturriaga1, J M Díaz-Mínguez, E P Benito
1Departamento de Microbiología y Genética, Facultad de Biología, Universidad de Salamanca, Spain.
Abstract:
The leu1 gene of Phycomyces blakesleeanus was isolated within a HindIII-HindIII genomic DNA fragment by heterologous hybridization screening of a cosmid library, making use of the Mucor circinelloides leuA gene as a probe. The complete nucleotide sequence of this fragment reveals a single 2070 bp ORF with no introns, which presents at least 68% homology with that of the leuA gene. The P. blakesleeanus leu1 gene has also been expressed in the M. circinelloides mutant R7B (leu-), which was used to isolate the leuA gene by complementation. The homology with other known sequences shows that the leu1 gene encodes the P. blakesleeanus alpha-IPM (isopropylmalate) isomerase.