Genomic deletions in Aureobasidium pullulans by an AMA1 plasmid for gRNA and CRISPR/Cas9 expression

Table 1 Number of transformants obtained on indicated selection media

Transformation¹	Plasmid² (µg)	V proto³ (mL)	Hygromycin B colonies⁴ (CFU in colonies/µg DNA)⁵		5-FOA⁸
Transformation¹	Plasmid² (µg)	V proto³ (mL)	Fresh⁶	Frozen⁷	5-FOA⁸
1	2	5.2	NT	150 (75)	105 out of 126 (83%)
1	5	5.2	NT	250 (50)	105 out of 126 (83%)
2	2	0.2	0	0	33 out of 33 (100%)
2	2	0.5	10 (52)	5 (26)
2	2	4.5	290 (168)	37 (21)
2	5	0.2	0	0
2	5	0.5	47 (98)	7 (15)
2	5	4.5	572 (132)	116 (27)

NT Not tested
¹Number of the transformation
²Quantity of plasmid DNA added during the transformation
³Volume of the used protoplast solution plated with agar (the total volume of the solution is 5.2 mL)
⁴Number of colonies on the hygromycin B selection plate and
⁵CFU in colonies per µg DNA given in brackets and calculated based on Eq. 1
⁶The protoplasts used for the transformation were freshly prepared
⁷The protoplasts used were thawed
⁸Number of colonies on the 5-FOA selection plate after transfer from the hygromycin B plate. The results are expressed in number of colonies growing out of the total number of colonies transferred, and the representative percentage is also given

ISSN: 2054-3085