Recombinant bacterial artificial chromosomes

A Bacterial Artificial Chromosome (BAC) is a vector that allows the propagation of larger exogenous DNA fragments, up to several hundred kb. BACs are propagated in recombination-deficient strains of E. coli. They are more stable and easier to handle than yeast artificial chromosomes (YACs). 

The principles that govern the delivery of recombinant DNA in clonable form to a host cell, and its subsequent amplification in the host, are well illustrated by considering three popular cloning vectors commonly used in experiments with E. coli—plasmids, bacteriophages, and bacterial artificial chromosomes—and a vector used to clone large DNA segments in yeast. 

Messerle M, Hahn G, Brune W, Koszinowski UH (2000) Cytomegalovirus bacterial artificial chromosomes a new herpesvirus vector approach. Adv Virus Res 55 463 -478 Mocarski-ES J, Kemble GW (1996) Recombinant cytomegaloviruses for study of replication and pathogenesis. Intervirology 39 320 330... 

Key words Recombineering, Hit and Fix method. Oligonucleotide, Bacterial artificial chromosome, Point mutation... 

Muyrers JP, Zhang Y, Testa G, Stewart AF (1999) Rapid modification of bacterial artificial chromosomes by ET-recombination. Nucleic Acids Res 27 1555-7. 

Various bacterial plasmids, bacteriophages, and yeast artificial chromosomes are used as cloning vectors (Brown, 2001). At the heart of the general approach to generating and propagating a recombinant DNA molecule is a set of enzymes which synthesize, modify, cut, and join DNA molecules. 

A major advance in replication-defective adenoviruses is the development of methods for creating recombinant viruses from distinct molecular clones in which the entire genome of the vector has been subcloned into a prokaryotic or invertebrate host. This has now been accomplished in a number of systems including yeast artificial chromosomes (21), cosmids (22), and bacterial plasmids (23-25). The subsequent discussion will focus on methods in which the adenoviral genome is subeloned in its entirety into bacterial plasmids,... 

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