Figure 2. Gene expression ratio for ovalbumin transcript in
DF1+/OVA Pro ∆ cells.
- Up- regulation of the ovalbumin mRNA in DF1 +/OVA Pro
∆cells were assessed by RT-qPCR. Upon deletion of the distal
ovalbumin promoter, an increased level of expression of ovalbumin was
determined (DF1 ∆). The
transcript levels of ovalbumin
for these samples are ~104 fold
higher than the ovalbumin transcript levels in the wild-type DF1 (WT
DF1). The transcript levels for ovalbumin in the hormonally-activated
tissue of magnum (from 35-week old laying hen) shows the highest level
of expression. Gene expression ratio for the Ovalbumin gene
over Gapdh has been calculated by the Pfaffl method of relative
quantification.
- Agarose gel (2%) electrophoresis for analysis of the RT-PCR products
amplified by primers P4 and P5 (for ovalbumin, Figure 1 and Table 2),
and P8 and P9 (for Gapdh , Table 2). The expected amplicon size
for ovalbumin and Gapdh are 179 bp and 187 bp, respectively.
WT, wild type; DF1 ∆, distal ovalbumin promoter
knockout DF1 cells (DF1 +/OVA Pro ∆); M, DNA size
marker; NTC, no template control. RT, reverse transcriptase.