Figure 3: Molecular analysis confirming the integration and transcription of HVA1 transgene in plants. (a) PCR results of T3 transgenic plants. The expected band size is 670 bp; (b) Southern blot hybridization showing integration of HVA1 gene digested with BamH1. The results indicate that there are two copies of transgene in all varieties. (c) RT-PCR of HVA1 expression for T2 transgenic plants. Like in PCR, here the expected band size is 670 bp for HVA1. Below the RT-PCR is the cDNA loading control showing the expression of ubiquitin with the expected band size of 450 bp. (d) Northern blot analysis also confirmed the transcription of HVA1 gene in Sedona and Matterhorn. Wt: wild type shows no transgene integration; C. RNA transcription analysis of HVA1 gene in T3 transgenic plants. Mat: “Matterhorn” and Sed: “Sedona” showed some expression. The remaining lanes, Wt: wild type, Mon: “Montcalm” and Con: “Condor” showed no transcriptions.