TABLE 2

Comparison of transformation strategies

Transformation strategyNo. of protoplasts usedAmt of linearized DNA used (μg)No. of transformants obtained (mean ± SD)Rate of positive transformants (%, mean ± SD)
Gene deletions using hyg as markera1073020 ± 263 ± 12
Gene integration using split markerb107503 ± 1100
Gene integration using full-length markerc107504 ± 186 ± 7
Ectopic gene integration (cotransformation)d1075 (target gene) and 1 (marker gene)e48 ± 1276 ± 14
  • a Deletion of asl1 and hah1 in this study.

  • b Integration of the EYFP gene upstream of asl1 using a partial asl1 expression cassette as the marker in this study.

  • c Integration of the EYFP gene upstream of pyr4 using a full-length pyr4 expression cassette as the marker in this study.

  • d Average results from our laboratory.

  • e Circular plasmids were used.