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Figure 5 | Genetic Vaccines and Therapy

Figure 5

From: Human cytomegalovirus plasmid-based amplicon vector system for gene therapy

Figure 5

Tn9-8-gpt was rescued from concatamers following serial passage in HF cells. (A) Tn9-8-gpt was passaged in HF cells and monomer units were recovered by linearizing concatemeric DNA from serial passage 3 with HindIII and cloning in bacteria (lanes 2 and 4) and also by linearizing passage 2 DNA with PstI and cloning in bacteria (lane 3). Lane 1 represents the unpassaged clone for comparison. Following rescue in bacteria, DNA was prepared and cut with NcoI. Fragments were separated on an agarose gel and visualized with ethidium bromide staining (left panel). The gel was transferred to a nylon membrane and probed with an a sequence specific probe (right panel). (B) Fragments from an NaeI digest were also separated on an agarose gel and visualized as in panel A. The gel was transferred to a nylon membrane and probed with an a sequence specific probe. Lane 5 shows a 100 bp ladder. Lanes 3 and 4 show a ca. 800 bp fragment that hybridizes to a sequences.

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