Vélez Pérez, Lisandra

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    Replication of the 3' LUIII 3' subgenomic clone in human cells
    (2004) Vélez Pérez, Lisandra; Diffoot Carlo, Nanette; College of Arts and Sciences - Sciences; Ríos Velázquez, Carlos; Montalvo Rodríguez, Rafael; Pastrana Ríos, Belinda; Department of Biology; Cádiz, Mayra E.
    Parvoviruses are small, non-enveloped, icosahedral viruses that contain a linear, single-stranded DNA genome of approximately 5000 bp. Parvovirus LuIII is an autonomous parvovirus that shares up to 80% of sequence homology with the rodent parvovirus MVM and H1, but encapsidates equal amounts of plus and minus strand, identical to AAV and B19. To study the possibility that the termini determine the strand to be encapsidated, a plasmid containing two copies of the left end termini was initially constructed. However, this plasmid known as 3' LuIII 3' has an insertion of unknown origin between the two termini. To eliminate this stuffer sequence, three different strategies were followed. The first two strategies consisted in eliminating the stuffer sequence by digesting the plasmid independently with PmeI and MluI enzymes, and then re-circulating it, and the third strategy consisted in constructing the recombinant molecule de novo. Irrespective of the strategy used, the desired 3' LuIII 3' clone was not obtained. To determine if the original clone 3' LuIII 3', was able of replicating in eukaryotic cells, the plasmid was co-transfected into HeLa cells with the infectious clone pGLu883ΔXba. No evidence of 3' LuIII 3' replication was observed.