Publication:
Study of an ARS-like function of map units 88-100 of parvovirus LUIII
Study of an ARS-like function of map units 88-100 of parvovirus LUIII
dc.contributor.advisor | Diffoot-Carlo, Nanette | |
dc.contributor.author | De Jesús-Maldonado, Idaris | |
dc.contributor.college | College of Arts and Sciences - Sciences | en_US |
dc.contributor.committee | Gill-Eccles, John | |
dc.contributor.committee | Massol-Deyá, Arturo | |
dc.contributor.department | Department of Biology | en_US |
dc.contributor.representative | Laboy, Jorge | |
dc.date.accessioned | 2018-08-09T14:30:47Z | |
dc.date.available | 2018-08-09T14:30:47Z | |
dc.date.issued | 2004 | |
dc.description.abstract | 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. | en_US |
dc.description.abstract | La síntesis de la hebra negativa por el parvovirus LuIII requiere la replicación del palindrome izquierdo de la hebra positiva de ADN. Un origen de replicación que no a sido identificado debe estar presente en el extremo derecho de la hebra negativa. El parvovirus LuIII contiene una secuencia única rica en A/T que comparte homología con las secuencias de replicación autónoma encontradas en levaduras. Para evaluar si la región rica en A/T y las secuencias río abajo de ésta son responsables de replicación autónoma, los nucleotidos 4527 a 5135 de LuIII fueron clonados (pUraLu88-100) en un vector que contenía el gene URA3 como un marcador auxotrófico. pUra-Lu88-100 fue transformado en el S. cerevisiae (SEY S288C, ura3 -) por el método de electroporación. El ADN del plásmido fue aislado de las colonias resultantes y transformado posteriormente en células del E. coli DH5α. Análisis realizados al plásmido rescatado de la levadura indican que el clon pUra- Lu88-100 fue capaz de existir en un estado libre en S. cerevisiae, sugiriendo que los elementos requeridos para llevar a cabo replicación autónoma limitada similar a la descrita para ARS están presentes en el terminal derecho del genoma viral de LuIII. | en_US |
dc.description.graduationYear | 2004 | en_US |
dc.identifier.uri | https://hdl.handle.net/20.500.11801/800 | |
dc.language.iso | en | en_US |
dc.rights.holder | (c) 2004 Idaris De Jesús-Maldonado | en_US |
dc.rights.license | All rights reserved | en_US |
dc.subject | Parvovirus LuIII | en_US |
dc.subject | A/T-rich sequence | en_US |
dc.subject | Saccharomyces cerevisiae (SEY S288C,ura3-) | en_US |
dc.subject | Echerichia coli DH5α cells | en_US |
dc.subject | LuIII viral genome | en_US |
dc.subject.lcsh | Parvoviruses | en_US |
dc.subject.lcsh | Viral genomes | en_US |
dc.subject.lcsh | Escherichia coli | en_US |
dc.subject.lcsh | Nucleotide sequence | en_US |
dc.title | Study of an ARS-like function of map units 88-100 of parvovirus LUIII | en_US |
dc.type | Thesis | en_US |
dspace.entity.type | Publication | |
thesis.degree.discipline | Biology | en_US |
thesis.degree.level | M.S. | en_US |
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