A versatile 2A peptide-based bicistronic protein expressing platform for the industrial cellulase producing fungus, Trichoderma reesei

Table 1 PCR analysis of selected C2G transformants along with their protein expression and fluorescence intensity measurements

Transformants	Fluorescence intensity	Western blot analysis		PCR analysis
Transformants	Fluorescence intensity	Cel7A	eGFP	PCR-1	PCR-2	PCR-3
A1	8393	+	+	+	+	+
C2	16,062	+	+	+	+	+
C4	2462	+	+	+	+	+
C5	17,975	+	+	+	+	+
D1	4358	+	+	+	+	+
D3	7360	+	+	+	+	+
A2	1976	−	−	−	+	−
A4	2736	−	−	+	+	+
AST1116	2130	−	−	−	–	−
JLT102	2088	+	−	−	–	−
SV001	ND	+	+	−	+	+
SV002	ND	+	+	−	+	+
SV004	10,539	+	+	+	+	+

Fluorescent intensities are shown in arbitrary fluorescence units (FLU)
+ and − under western blot and PCR analysis columns represent presence or absence of the protein and gene, respectively, Faint represents very low, but detectable levels of the respective protein, ND denotes no data available, Controls JLT102A (AST1116 expressing native Cel7A under the eno promoter), AST1116 (Cel7A-deleted T. reesei QM6a strain), SV001 and SV002 (AST1116 expressing eGFP-2A-Cel7A; colonies C2-1 and D2-1 in Fig. 3a), and SV004 (AST1116 expressing Cel7A-2A-eGFP; colony A5-4 in Fig. 3a)

ISSN: 2731-3654