Cloning of the Rhizopus niveus pyr4 gene and its use for the transformation of Rhizopus delemar
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- Horiuchi, H., Takaya, N., Yanai, K. et al. Curr Genet (1995) 27: 472. doi:10.1007/BF00311218
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We have cloned a pyr4 gene encoding orotidine-5′-monophosphate decarboxylase of the filamentous fungus Rhizopus niveus. The pyr4 gene of R. nivens has an open reading frame composed of 265 amino-acid residues and has two putative introns. We have also isolated a pyr4 mutant of Rhizopus delemar from 5-fluoroorotic acid-resistant mutants and transformed it with the pyr4 gene of R. niveus as a selectable marker. Introduced DNA was integrated into the chromosome in a multiple tandem array. The mitotic stability of the introduced DNA was increased by a repeated sporulation process. The expression of the Escherichia coli β-glucuronidase gene in R. delemar was successfully obtained under the control of the pgk2 gene promoter of R. niveus by co-transformation with the pyr4 gene.