De Jesús-Maldonado, Idaris

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Study of an ARS-like function of map units 88-100 of parvovirus LUIII
(2004) De Jesús-Maldonado, Idaris; Diffoot-Carlo, Nanette; College of Arts and Sciences - Sciences; Gill-Eccles, John; Massol-Deyá, Arturo; Department of Biology; Laboy, Jorge
The synthesis of minus strand by parvovirus LuIII requires replication from the left palindrome of the plus strand DNA. An origin of replication yet to be identified must be present at the right end of the minus strand, required for the synthesis of plus strand. Parvovirus LuIII contains a unique A/T-rich sequence that shares a sequence homology with the autonomously replicating sequences (ARS) found in yeast. To test if the A/T rich region and sequences downstream the A/T sequence are responsible for autonomous replication, nucleotides 4527 to 5135 of LuIII were cloned (pUraLu88- 100) into a vector containing the URA3 gene as an auxotrophic marker. pUra-Lu88- 100 was transformed into Saccharomyces cerevisiae (SEY S288C,ura3-) by the electroporation method. Plasmid DNA was isolated from the resulting colonies and transformed into Echerichia coli DH5α cells. Analysis performed to the plasmid rescued from yeast indicate that pUra-Lu88-100 was capable of existing in a free state in S. cerevisiae, suggesting that the elements required for a limited ARS-like function are present in at the right end of the LuIII viral genome.