转座标签法构建禾谷镰刀菌插入突变体

he objective of the present study was to obtain the inserted mutants in Fusarium graminearum. We co-transformed the plasmids, which contained transposon element mimp1 and transposase impala E respectively, into the niaD mutant by the PEG-mediated protoplast transformation. Then, we selected the hygromycin-resistant transformants and screened these transformants by PCR and southern blot analysis to get the original strain. Five hundred mutants were obtained by cultivated the original strain 2047 on MM medium which contained nitrate nitrogen as the sole nitrogen source. Thirty six inserted sites which identified by Tail-PCR and sequencing scattered randomly on four chromosomes. Among them, twenty two mutants with its mimp1 inserted into promoter regions were identified. Random distribution in four chromosomes and a significant bias towards promoter regions of mimp1-tagging system has advantages to select and identify genes in Fusarium graminearum.