Methods on erg9 gene deletion in Schizosaccharomyces pombe

Abstract

Gene modification to key enzymes in biosynthesis pathway becomes familiar technology which can increase efficiency of industrial strains. In the current study, we established a desirable deletion approach based on homologous recombination. It was simple and available for spn5 gene deletion. Also, four kinds of erg9 gene deletion cassettes were constructed in similar manner. The lengths of flanking sequences for homologous integration were from 59 bp to more than 1 Kb, and three selective markers (G418, Zeocin and LEU2) were employed in these cassettes. However, Δerg9 mutant was not obtained in this work. The possible reason is that erg9 gene is an essential gene and a single copy in its chromosome. Deficiency of erg9 gene resulted in nonviability to the cells. In addition, LEU2 marker caused unspecific integration and the difficulty of screening mutants is increased during deletion procedures. This work provides important evidence to modify industrial strains by using deletion technologies. © 2009 Curtin University of Technology and John Wiley & Sons, Ltd.