Fig.3-2 Construction diagram of MareA gene
overexpression vector
The RT-qPCR was performed using the cDNA of C100 and ΔMareA as
templates, with qMareA-F and qMareA-R serving as primers. The expression
of the MareA gene in both strains was compared through
calculation. By comparing the expression levels of the MareA gene
in C100 and ΔMareA strains, it was observed that the gene could
be expressed normally in the original strain C100, while its expression
in the knockout strain ΔMareA approached zero. This suggests that
there is almost no transcriptome of the MareA gene in
ΔMareA, further confirming successful knockdown of theMareA gene.
The cDNAs of C100 and OE-MareA were used as templates for
RT-qPCR, with qMareA-F and qMareA-R serving as primers. The expression
of the MareA gene in both strains was compared through
calculation. The relative expression level of the MareA gene in
the overexpression strain OE-MareA was found to be 3.41 times
higher than that in the original strain C100, indicating successful
insertion of the MareA gene fragment into the genome of monascus
C100 and successful screening of the overexpression transformantOE-MareA . The results are presented in Figure 3-3.