<p style="text-align: justify;">The CZW-3 squash line was produced using an <i>Agrobacterium</i>-meditated transformation of the parental variety YS20 via a protocol in which the transfer-DNA (T-DNA) region of the bacterial tumour inducing (Ti) plasmid was modified using recombinant DNA techniques to contain the coat protein (CP) encoding sequences from CMV, ZYMV and WMV2. During transformation, the T-DNA portion of the plasmid was transferred into the plant cells and stably integrated into the plant&#39;s genome, and there was no incorporation of backbone plasmid sequences outside of the T-DNA region.<br />
The constitutive expression of the CP genes was regulated by the 35S promoter and termination sequences from cauliflower mosaic virus (CaMV). In addition, all three CP genes were fused to the 5&#39; untranslated sequence from CMV to enhance translation of the transgene mRNA.</p>
