Construction and characterization of bacterial artificial chromosomes containing HSV-1 strains 17 and KOS.

Journal of Virological Methods
William W GieraschDavid A Leib

Abstract

Bacterial artificial chromosomes (BACs) were constructed containing full-length, infectious DNA of HSV-1 strains 17 and KOS. To generate BACs without altering viral genes, sequences required for selection and propagation of the BAC were placed between the U(L)37 and U(L)38 genes, and flanked by LoxP sites. The system was tested by studying multiple properties of these HSV-1 BAC constructs in vitro and in vivo following propagation in bacteria, virus reconstitution from HSV-BAC DNA in eukaryotic cells, and Cre-recombinase-mediated excision of the BAC backbone. Based on in vitro growth in mouse embryo fibroblasts and in vivo growth in mouse corneas and trigeminal ganglia, the strain KOS BAC-derived virus behaved similarly to wild-type. Small changes in neurovirulence were, however, observed. The strain 17 BAC-derived virus exhibited modest decreases in growth and virulence compared to wild-type. Modest differences were observed in reactivation from latency with both strain KOS and 17 BAC-derived viruses. In addition, the system was further validated by performing mutagenesis of the BACs by allelic exchange in E. coli. These BACs are suitable for the rapid generation of recombinant viruses for pathogenesis and other studies, but a...Continue Reading

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Citations

Aug 15, 2013·Analytical Chemistry·Shaohui HuHeng Zhu
Oct 3, 2012·Proceedings of the National Academy of Sciences of the United States of America·Matthew P TaylorLynn W Enquist
Nov 17, 2010·Antimicrobial Agents and Chemotherapy·Jocelyne Piret, Guy Boivin
Apr 11, 2008·Journal of Virology·Tracy Jo PasiekaDavid A Leib
Aug 24, 2007·Journal of Virology·Kelly Elizabeth CollerGregory Allan Smith
Apr 2, 2010·Journal of Virology·Holly A SaffranJames R Smiley
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Nov 20, 2009·Journal of Virology·Sarah Elizabeth Antinone, Gregory Allan Smith
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Sep 10, 2014·Molecular Therapy : the Journal of the American Society of Gene Therapy·Lucia MazzacuratiPaola Grandi

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