Construction of deletion mutants for three proteasome protein coding genes and characterization of their phenotypes in Neurospora crassa

Abstract:

Abstract: [Objective] In this study, we generated knockout strains of rpn4, rpn7, and rpn10 genes respectively in Neurospora crassa, and studied their phenotypes to uncover their functions in the 26 S protesome pathway. [Methods]We used gene-replacement strategy to make knockout strains. After electrotransformation, the resulted transformants were examined by PCR. To get homokaryotic mutants, the heterokaryotic strains were crossed with the wild-type strain. The ascospores were germinated after heat shock, and the resulting strains were examined by PCR to confirm the homokaryotic strains. [Results] Homokaryotic rpn4 and rpn10 knockout strains, and heterokaryotic rpn7KO strains were obtained. [Conclusion] Compared to the wild-type strain, rpn7KO strains exhibited severe defects in hyphal growth and conidial formation; rpn4KO strain had defect in hyphal growth too, but displayed normal conidiation; the growth and developmental phenotypes of rpn10KO strain were slightly affected. Taken together, these data demonstrated that these three genes were important for the growth and developmental phenotypes of Neurospora crassa.