Restriction digests of the clone give the following sizes (kb): KpnI--2.9, 1.3; NotI--2.9, 1.3; XhoI--4.2. Restriction sites flanking the 2X35S/EPITOPE/NOS 3' cassette allow excision with either NotI for subcloning into the binary vector pART27 or KpnI for subcloning into the binary v ector pRD400. There are also unique SacI and XhoI sites for either directional subcloning or insertion of other plant transgenes before subcloning. The vector contains a specialized cassette allowing the in-frame N- or C-terminal fusion of a polymerase chain reaction (PCR)-amplified mature protein coding sequence from any gene to a sequence encoding an epitope of the influenza hemagglutanin a-chain. The vector was constructed using the pGEM-9Zf derivative pART7 from the vector PJIT166 and the 3' transcription terminator from the nopaline synthase (NOS) gene subcloned from the vector pCaGUS. |
