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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

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

Plasmid(s) ade8+-selected transformants collected on* Total transformants*
1st day 2nd day 3rd day 4th day
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)
  1. *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
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