Bacterial artificial chromosomes or BACS are circular DNA molecules which

contain a replicon that is based on the F factor. BACs have oriS and repE which
encode an ATP-driven helicase and parA, parB and parC for partitioning. The
original BAC vectors are 7.4 kb while others are 8-9kb in length. You can clone
80-300kb fragments in BACs. Generally, the host needs to be deficient in
homologous recombination machinery in order for the BAC to be stable (i.e.
recA-).

repE
for plasmid replication and regulation of copy number.
parA and parB
for partitioning F plasmid DNA to daughter cells during division and ensures stable
maintenance of the BAC.
A selectable marker
for antibiotic resistance; some BACs also have lacZ at the cloning site for blue/white
selection.
T7 & Sp6
phage promoters for transcription of inserted genes.

Bacterial artificial chromosome (BAC)

Bacterial artificial chromosomes (BACs) involve a cloning system that is derived from a
particular plasmid found in the bacterium Escherichia coli . The use of the BAC allows large
pieces of deoxyribonucleic acid (DNA ) from bacterial or non-bacterial sources to be
expressed in Escherichia coli. Repeated expression of the foreign DNA produces many
copies in the bacterial cells, providing enough material for analysis of the sequence of the
DNA. BACs proved useful in the sequencing of the human genome.
The BAC is based on a plasmid in Escherichia coli that is termed the F (for fertility) plasmid.
The F plasmid (or F factor) contains information that makes possible the process called
conjugation . In conjugation, two Escherichia coli bacteria can physically connect and an
exchange of DNA can occur.
A BAC contains the conjugation promoting genetic information as well as stretch of DNA
that is destined for incorporation into the bacterium. The foreign DNA (e.g., portion of human
genome) is flanked by sequences that mark the boundaries of the insert. The sequences are
referred to as sequence tag connectors. When the BAC becomes incorporated into the
genome of Escherichia coli the sequence tag connectors act as markers to identify the
inserted foreign DNA.
Using a BAC, large stretches of DNA can be incorporated into the bacterial genome and
subsequently replicated along with the bacterial DNA. In molecular biology terminology,
pieces of DNA that contain hundreds of thousands of nucleotides (the building blocks of
DNA) can be inserted into a bacterium at one time. As the process is done using different
sections of the foreign DNA, the amount of DNA that can be analyzed can be very large.
BACs were developed in 1992. Since then, their usefulness has grown immensely. The
primary reason for this popularity is the stability of the inserted DNA in the bacterial genome.

Because the inserted DNA remains in the bacterial genome during repeated cycles of
replication, the information is not lost. As well, the BAC can be sequenced using the normal
tools of molecular biology.
The most dramatic recent example of the power of BACs is their use by The Institute for
Genomic Research (TIGR ) in the technique of shotgun cloning that was employed in the
sequence determination of the human genome. Many fragments of the human genome could
be incorporated into BACs. The resulting "library" could be expressed in Escherichia coli
and the sequences determined. Subsequently, these sequences could be reconstructed to
produce the orderly sequence of the actual genome. This approach proved to be less
expensive and quicker than the method known as directed sequencing, where a genome was
sequenced in a linear fashion starting at one end of the genome.
The total number of fragments of the DNA from the human genome that have been expressed
in Escherichia coli by the use of BACs is now close to one million. In addition to the human
genome, BACs have also been used to sequence the genome of agriculturally important
plants such as corn and rice, and of animals such as the mouse.
With the realization of the sequence of the human genome, the use of BACs is becoming
important in the screening of the genome for genetic abnormalities.
Infectious disease
The genomes of several large DNA viruses and RNA viruses have been cloned as BACs.
These constructs are referred to as "infectious clones", as transfection of the BAC construct
into host cells is sufficient to initiate viral infection. The infectious property of these BACs
has made the study of many viruses such as the herpesviruses, poxviruses and coronaviruses
more accessible.[5][6][7] Molecular studies of these viruses can now be achieved using genetic
approaches to mutate the BAC while it resides in bacteria. Such genetic approaches rely on
either linear or circular targeting vectors to carry out homologous recombination.[8]