The invention discloses a method for
gene modification of
aureobasidium pullulans, which belongs to the technical field of
gene engineering, and comprises the following steps: determining a
malic acid synthase
gene, obtaining a
malic acid synthase gene segment, searching a
gene sequence of the
malic acid synthase in the disclosed
aureobasidium pullulans as a template on NCBI (National Center of
Biotechnology Information), and carrying out
gene modification on the malic acid synthase gene segment to obtain a malic acid synthase gene segment; the method comprises the following steps: respectively designing an upstream primer and a downstream primer, taking extracted
genome DNA of
aureobasidium pullulans M1 as a template, respectively carrying out PCR amplification and recombinant
plasmid pUC57-PMA-ms construction and
linearization treatment on the upstream primer and the downstream primer, and introducing the
plasmid into the
aureobasidium pullulans M1 to obtain an
aureobasidium pullulans recombinant strain; according to the method, the
polymalic acid synthetase gene (PMA-ms) on an
aureobasidium pullulans genome is knocked out from aureobasidium pullulans, the yield of
pullulan in the obtained recombinant
plasmid M1-2 is about 39.11 g / L, and compared with an original strain M1, the yield of M1-2 is increased by 9.79 g / L, and the practical application prospect of the method is improved.