Unit

UNIT 10.32 Use of Bacterial Artificial Chromosomes in Generating Targeted Mutations in Human and Mouse Cytomegaloviruses

  1. Eva Maria Borst,
  2. Corinna Benkartek,
  3. Martin Messerle

Published Online: 1 MAY 2007

DOI: 10.1002/0471142735.im1032s77

Current Protocols in Immunology

Current Protocols in Immunology

How to Cite

Borst, E. M., Benkartek, C. and Messerle, M. 2007. Use of Bacterial Artificial Chromosomes in Generating Targeted Mutations in Human and Mouse Cytomegaloviruses. Current Protocols in Immunology. 77:VII:10.32:10.32.1–10.32.30.

Author Information

  1. Institute of Virology, Hannover Medical School, Hannover, Germany

Publication History

  1. Published Online: 1 MAY 2007
  2. Published Print: MAY 2007

Abstract

Cloning of cytomegalovirus (CMV) genomes as bacterial artificial chromosomes (BAC) in E. coli and their manipulation using the techniques of bacterial genetics has greatly facilitated the construction of CMV mutants. This unit describes easily applicable procedures that allow rapid introduction of any kind of targeted mutation into BAC-cloned CMV genomes. Protocols for the reconstitution of virus from isolated BAC DNA, preparation of a virus stock, and isolation and characterization of viral DNA are also included. Special emphasis is laid on description of critical steps and thorough characterization of the altered BACs.

Keywords:

  • cytomegaloviruses;
  • bacterial artificial chromosomes;
  • mutagenesis