Current Genetics

, Volume 27, Issue 5, pp 472–478 | Cite as

Cloning of the Rhizopus niveus pyr4 gene and its use for the transformation of Rhizopus delemar

  • H. Horiuchi
  • N. Takaya
  • K. Yanai
  • M. Nakamura
  • A. Ohta
  • M. Takagi
Original Paper

Abstract

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.

Key words

Rhizopus niveus pyr4 Rhizopus delemar Transformation 

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Copyright information

© Springer-Verlag 1995

Authors and Affiliations

  • H. Horiuchi
    • 1
  • N. Takaya
    • 1
  • K. Yanai
    • 1
  • M. Nakamura
    • 1
  • A. Ohta
    • 1
  • M. Takagi
    • 1
  1. 1.Department of Biotechnology, Division of Agriculture and Agricultural Life SciencesThe University of TokyoTokyoJapan

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