## 2 Materials and Methods
### Western Blot  
Separation of proteins for Western blot analysis was performed by denaturing polyacrylamide gel electrophoresis according to the Laemmli method [13]. Plant extracts were leveled in protein content; 20 ug of total protein was applied to the lane. The transfer of proteins from the gel to a nitrocellulose membrane was carried out in a semi-dry blot system (Trans-Blot SD semi-dry transfer cell, Bio-rad, Hercules, CA, USA) at a current of 2 mA/cm2 for 30 min. We used primary polyclonal goat antibodies to recombinant interferon-gamma (R&D systems, Abingdon, UK) and secondary polyclonal anti-goat rabbit antibodies labeled with horseradish peroxidase (Agrisera, Vannas, Sweden). The development of the nitrocellulose membrane was carried out using 3,3^-diaminobenzidine. The color intensity and the area of interferon-gamma spots on the membrane were assessed using the ImageJ 1.41 software. The content of interferon was calculated in relative units, which were obtained by multiplying the area of the spot on the blot by the intensity of its color, and then the approximate content in ug was calculated relative to the commercial preparation of interferon-gamma, which was applied at a concentration of 1 ug per lane (positive control).  
\begin{table}
\begin{tabular}{c c c c} \hline \hline p35S (cauliflower mosaic & 35S\_F & CCTCCTCGGATTCCATTGCCC \\ \cline{2-3}  & virus promoter) & 35S\_R & GTCCATCTTTGGACCACTGTCGG \\ \cline{2-3}  & OCST (octopine synthase & OCS\_F & GCGAGAGCCCTATGCATGCATGC \\ \cline{2-3}  & gene terminator) & OCS\_R & GAAACCGGGAAGGATCTGAGC \\ \cline{2-3}  & pNOS (noplasmic synthase & NOS\_F & CGATGACGCGGAACAAGCCGT \\ \cline{2-3}  & gene promoter) & NOS\_R & GACCTTAGGGCACTTTGAACCGC \\ \cline{2-3}  & _NPT II_ (neomycin & NPT\_F & GTGTTCCGGTGTCAGGCAGG \\ \cline{2-3}  & phosphotransferase) & NPT\_R & CCCAAGGAACCCCCCTCTCG \\ \cline{2-3}  & RB and LB (right and left & RB\_in\_1 & GTTACCGGCCAATATATCCT \\ \cline{2-3}  & borders of T-DNA) & LB\_in\_1 & TGCACGATATATTTGGTGTGA \\ \hline \multirow{4}{*}{**RT-PCR**} & \multirow{2}{*}{_EF-1a_ (reference gene)} & EF1 & CAAGCGGCTATCAAGTAGC \\ \cline{2-3}  & & EF2 & TGTCCGAGCAGCATCAATCACA \\ \cline{2-3}  & _RISIFNG_ (recombinant & RTSIFNG-1 & AGGAGATATGGACATCATCAAAGCA \\ \cline{2-3}  & bovine gamma-interferon) & RTSIFNG-2 & AGTCGCTGGACCGGATTTGA \\ \hline \multirow{4}{*}{**FPNI-PCR**} & \multirow{4}{*}{T-DNA sequence near right border (RB)} & 35 Sm\_antiSP & CTGATCATAGGCGAGAATTAAAGGGA \\ \cline{2-3}  & & 35 Sm\_SP3 & TCGGAAAACCTGCTGCCA \\ \cline{2-3}  & T-DNA sequence near right border (RB) & 35 Sm\_SP2 & GCTCACTGCCCCGCTTCCCAG \\ \cline{2-3}  & & 35 Sm\_SP1 & AGTGTAAAGCCCTGGGGGTCCT \\ \cline{2-3}  & & NOST\_antiSP & CCTAGTTTGCCGCTATATTTGTGTTTCC \\ \cline{2-3}  & T-DNA DNA sequence near left border (LB) & NOST\_SP3 & GAAGCAGATCGTTCAAACATTTGGC \\ \cline{2-3}  & & NOST\_SP2 & TCGTTTTGGGTCTATCCACCGGT \\ \cline{2-3}  & & NOST\_SP1 & CCAACTGCCAAAATCATCCACACCGT \\ \hline \multirow{4}{*}{**Conformation of T-DNA insertion sites in the plant genome**} & \multirow{4}{*}{B6 line} & B6\_R1 & TAGCCAACATACCTCAATTAGAGCTTCCT \\ \cline{2-3}  & B6\_L2 & AATGGCCGAGAGTGAAGCA \\ \cline{2-3}  & & 311\_L1 & AGCACTTCCCAGGCGCATATTACC \\ \cline{2-3}  & & 311\_R1 & CAAATGAAGTTTTGCTAATGTATGGAAT \\ \hline \hline \end{tabular}
\end{table}
Table 1: Primers used in the process.