Figure 2. Gene expression ratio for ovalbumin transcript in DF1+/OVA Pro ∆ cells.
  1. 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.
  2. 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.