Báez Crespo, Alexa M.

Profile Picture

Filters

20111
Reset filters

Settings

Citations

Publication Search Results

Now showing 1 - 1 of 1
  • Thumbnail Image
    Publication
    Replication efficiency of the LuIII Parvovirus miinigenomes 3’LUIII3’ and 3’LUIII5’
    (2011) Báez Crespo, Alexa M.; Diffoot Carlo, Nanette; College of Arts and Sciences - Sciences; Montalvo Rodríguez, Rafael; Ríos Velázquez, Carlos; Department of Biology; Parés Matos, Elsie I.
    Parvoviruses are currently being studied for use as possible genetic vectors in gene therapy. They are small, non-enveloped viruses of icosahedral structure, and possess a single-stranded DNA genome of approximately 5,000 base pairs that replicate only during the S-phase of the host cell. The viral replication strategy resembles the rolling circle replication model requiring the viral protein NS1. This protein functions as a site specific DNA binding protein, thus, binding the viral origin and initiating self-replication process. Studies have shown that genetic constructs with two viral terminal palindromic structures are sufficient for autonomous replication of the molecule in the presence of NS1. To compare the replication of a minigenome containing two left termini with that of one containing a left and right palindrome of LuIII the DsRed gene from the pCMV-DsRed (Clontech) was inserted between the hairpins of both vectors. The red fluorescent protein (DsRed) allowed monitoring of the replication of these constructs. The results show that LuIII 3'-DsRed-3’ and LuIII 3'-DsRed-5’ were both capable of expressing fluorescence in the cells over time, thus, indicating that both recombinant plasmids were capable of self replication. The results also suggest that replication of LuIII 3'-DsRed-3’ was more efficient than LuIII 3'- DsRed-5’. This result suggests that a LuIII genetic vector containing two copies of the 3’ hairpin (identical palindromic sequences) may be more efficient than a vector containing the LuIII 3’ and 5’ palindromes. This research has been helpful in providing insight about the replication efficiency expected, based on the response of the 3’ and 5’ terminal hairpin arrangements in genetic vectors used in gene therapy.