Document Detail


Cloning of the genomes of human cytomegalovirus strains Toledo, TownevarRIT3, and Towne long as BACs and site-directed mutagenesis using a PCR-based technique.
MedLine Citation:
PMID:  12667824     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The 230-kb human cytomegalovirus genome is among the largest of the known viruses. Experiments to determine the genetic determinants of attenuation, pathogenesis, and tissue tropism are underway; however, a lack of complete sequence data for multiple strains and substantial problems with genetic instability during in vitro propagation create serious complications for such studies. For example, recent findings suggest that common laboratory strains Towne and AD169 passaged in cultured human fibroblasts are missing up to 15 kb of genetic information relative to clinical isolates. To establish standard, genetically stable genomes that can be sequenced, disseminated, and repeatedly reconstituted to produce virus stocks, we have undertaken to clone two variants of Towne, designated Towne(long) and Towne(short) (referred to as TownevarRIT3) (A., Proc. Natl. Acad. Sci. USA 98, 7829-7834), and the pathogenic strain Toledo into bacterial artificial chromosomes (BACs). We further demonstrate the ease with which mutagenesis can be achieved by deleting 13.5 kb from the Toledo genome using a PCR-based technique.
Authors:
Gabriele Hahn; Dietlind Rose; Markus Wagner; Sylvia Rhiel; Michael A McVoy
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Virology     Volume:  307     ISSN:  0042-6822     ISO Abbreviation:  Virology     Publication Date:  2003 Mar 
Date Detail:
Created Date:  2003-04-01     Completed Date:  2003-05-16     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0110674     Medline TA:  Virology     Country:  United States    
Other Details:
Languages:  eng     Pagination:  164-77     Citation Subset:  IM    
Affiliation:
Max von Pettenkofer Institut, Abteilung für Virologie, Ludwig-Maximilians-Universitt München, Pettenkoferstrasse 9A, D-80336 Munich, Germany. ghahn@m3401.mpk.med.uni-muenchen.de
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MeSH Terms
Descriptor/Qualifier:
Cell Line
Chromosomes, Artificial, Bacterial
Cloning, Molecular / methods
Cytomegalovirus / classification*,  genetics*
Genome, Viral*
Humans
Mutagenesis, Site-Directed
Polymerase Chain Reaction / methods
Restriction Mapping

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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