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Updated: Sep 25, 2026
Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning
Published on: February 5, 2021
Cloning, characterization and heterologous expression of the Blakeslea trispora gene encoding
María Dolores Quiles-Rosillo1, Rosa María Ruiz-Vázquez, Santiago Torres-Martínez
1Departamento de Genética y Microbiología, Facultad de Biología, Universidad de Murcia, 30071, Murcia, Spain.
Abstract:
The pyrG gene of the fungus Blakeslea trispora, encoding orotidine-5'-monophosphate decarboxylase (OMPD) enzyme, was cloned by heterologous hybridization of a genomic library with the Mucor circinelloides pyrG gene. The deduced amino acid sequence of the B. trispora pyrG gene is highly similar to the OMPD from other organisms. Hybridization analyses revealed that the only copy of this gene present in the genome of B. trispora is constitutively expressed. Heterologous complementation of a mutant of M. circinelloides deficient in OMPD activity with the B. trispora pyrG gene and promoter sequence confirmed the function of this gene. This functional complementation demonstrates that heterologous expression in M. circinelloides might be used to investigate the function of genes of B. trispora.
