Construction, expression and enzymatic activity analysis of AUR1 eukaryotic expression vector of Botrytis cinerea / 生物工程学报

ABSTRACT

In order to study the expression and the activity of inositol phosphorylceramide synthase (BcAUR1 gene) in Botrytis cinerea, we amplified BcAUR1 by RT-PCR from Botrytis cinerea, using the special primers with FLAG and BamH I/Xho I restriction sites. Recombinant pYES2-BcAUR1 was constructed to transform into Saccharomyces cerevisae deltayorl by LiAC. The expression of inositol phosphorylceramide (IPC) synthase and its activity were detected by Western blotting and HPLC, respectively. The results show that pYES2-BcAUR1 could express in uracil mutant deltayorl of Saccharomyces cerevisae. IPC synthase enzyme activity of pYES2-BcAUR1 transformants significantly increased and was approximately double than no-load BcAUR1 transformants. The low concentration of Aureobasidin A could inhibit growth of no-load BcAUR1 transformants, but pYES2-BcAUR1 transformants could resist fungal growth inhibition which was induced by Aureobasidin A.