Development of a pyrG Mutant of Aspergillus oryzae Strain S1 as a Host for the Production of Heterologous Proteins

作者：Ling Selina Oh Siew; Storms Reginald; Zheng Yun; Rodzi Mohd Rohaizad Mohd; Mahadi Nor Muhammad; Illias Rosli Md; Murad Abdul Munir Abdul; Abu Bakar Farah Diba^*

来源：The Scientific World Journal, 2013, 2013: 634317.

DOI：10.1155/2013/634317

The ease with which auxotrophic strains and genes that complement them can be manipulated, as well as the stability of auxotrophic selection systems, are amongst the advantages of using auxotrophic markers to produce heterologous proteins. Most auxotrophic markers in Aspergillus oryzae originate from chemical or physical mutagenesis that may yield undesirable mutations along with the mutation of interest. An auxotrophic A. oryzae strain S1 was generated by deleting the orotidine-5%26apos;-monophosphate decarboxylase gene (pyrG) by targeted gene replacement. The uridine requirement of the resulting strain GR6 pyrG Delta 0 was complemented by plasmids carrying a pyrG gene from either Aspergillus nidulans or A. oryzae beta-Galactosidase expression by strain GR6 pyrG Delta 0 transformed with an A. niger plasmid encoding a heterologous beta-galactosidase was at least 150 times more than that obtained with the untransformed strain. Targeted gene replacement is thus an efficient way of developing auxotrophic mutants in A. oryzae and the auxotrophic strain GR6 pyrG Delta 0 facilitated the production of a heterologous protein in this fungus.

• 出版日期2013