Publication:
Replication of the 3' LUIII 3' subgenomic clone in human cells

dc.contributor.advisor Diffoot Carlo, Nanette
dc.contributor.author Vélez Pérez, Lisandra
dc.contributor.college College of Arts and Sciences - Sciences en_US
dc.contributor.committee Ríos Velázquez, Carlos
dc.contributor.committee Montalvo Rodríguez, Rafael
dc.contributor.committee Pastrana Ríos, Belinda
dc.contributor.department Department of Biology en_US
dc.contributor.representative Cádiz, Mayra E.
dc.date.accessioned 2018-08-09T14:38:47Z
dc.date.available 2018-08-09T14:38:47Z
dc.date.issued 2004
dc.description.abstract 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. en_US
dc.description.abstract Los parvovirus son pequeños virus icosaédricos sin envoltura que contienen un DNA lineal de hebra sencilla de aproximadamente 5000 pb. El virus LuIII es un parvovirus autónomo que comparte hasta un 80% de homología con los parvovirus autónomos MVM y H-1, pero éste encapsida cantidades iguales de hebras positiva y negativa. Este patrón es idéntico a AAV y B-19. Para estudiar la posibilidad de que los terminales estén determinando la polaridad de la hebra que se encapsida, un plásmido que contiene dos copias del terminal 3' fue construido. Sin embargo, este plásmido llamado 3' LuIII 3', tiene una inserción de origen desconocido entre los dos terminales. Tres estrategias distintas se llevaron a cabo para eliminar esta secuencia. Las primeras dos estrategias consistieron en eliminar esta secuencia desconocida cortando el plásmido por separado con las enzimas PmeI y MluI. La tercera estrategia consistió en construir la molécula recombinante de novo. Independientemente de la estrategia utilizada, no se obtuvo un clon 3' LuIII 3' como fuera deseado. Para determinar si el clon original 3' LuIII 3' era capaz de replicarse en células eucarióticas, el plásmido fue co-transfectado con el clon infeccioso pGLu883ΔXba en células HeLa, no observándose evidencia de replicación para el plásmido 3' LuIII 3'. en_US
dc.description.graduationYear 2004 en_US
dc.identifier.uri https://hdl.handle.net/20.500.11801/810
dc.language.iso en en_US
dc.rights.holder (c) 2004 Lisandra Vélez Pérez en_US
dc.rights.license All rights reserved en_US
dc.subject Parvoviruses en_US
dc.subject Icosahedral viruses en_US
dc.subject Parvovirus LuIII en_US
dc.subject Autonomous parvovirus en_US
dc.subject Rodent parvovirus MVM and H1 en_US
dc.subject Plasmid known as 3' LuIII 3' en_US
dc.subject.lcsh Parvoviruses en_US
dc.subject.lcsh Genomes en_US
dc.subject.lcsh Molecular cloning en_US
dc.subject.lcsh Recombinant molecules en_US
dc.title Replication of the 3' LUIII 3' subgenomic clone in human cells 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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