Selection markers for transformation of the sequenced reference monokaryon Okayama 7/#130 and homokaryon AmutBmut of Coprinopsis cinerea

Table 2 Transformations of C. cinerea OK130 (ade8-1) with ade8⁺-vector pCcAde8 alone or, using same batches of protoplasts, in combination with various pYSK7 laccase gene derivatives

Plasmid(s)	ade8⁺-selected transformants collected on*				Total transformants*
Plasmid(s)	1st day	2nd day	3rd day	4th day	Total transformants*
Experiment 1: Laccase overexpression
pCcAde8	17 (1)	15	7	2	41
pCcAde8 + pYSK7	26 (8)	20 (13)	7 (3)	7 (1)	60 (25)
pCcAde8 + pYSK-lcc5	14 (2)	27 (8)	25 (5)	10 (5)	76 (20)
pCcAde8 + pYSK-lcc9	10 (4)	23 (10)	23 (5)	8 (0)	64 (19)
Experiment 2: Laccase silencing
pCcAde8	17	20	12	6	55
pCcAde8 + pYSK-lcc9-antisense-1	5 (2)	7 (2)	12 (6)	6 (4)	30 (14)
pCcAde8 + pYSK-lcc9-antisense-2	2 (1)	9 (5)	17 (12)	8 (6)	36 (24)

*Data in brackets of experiment 1 indicate number of clones with > sixfold increased levels of laccase as detected by activity assay in liquid fermentation and native-PAGE; data in brackets of experiment 2 indicate clones with 2- to 11-fold (2^‒ΔΔCT) decreases in lcc9 mRNA transcriptional levels as detected by qRT-PCR

ISSN: 2054-3085