|
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)
|
- *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
