YAC and BAC cloning systems. The general steps involved in the construction of yeast artificial chromosome (YAC, left) and bacterial artificial
chromosome (BAC, right) clones are summarized. Specifically, high-molecular-weight source DNA (e.g., human DNA) is carefully prepared, partially
digested with a restriction enzyme, and size selected to yield large DNA fragments (e.g., typically about 200 to 1000 kb for YACs and about 100 to 300 kb
for BACs). Appropriate vector sequences are then ligated to the size-selected, insert DNA. For YACs, this consists of two vector arms that together contain
all the structural elements necessary for the propagation of a chromosome in yeast (see Green et al.76 for details). For BACs, this consists of a single
vector fragment that contains a suitable antibiotic-resistance gene (see Birren et al.114 for details). The ligated DNA is then transformed into appropriately
Source: General Themes, The Online Metabolic and Molecular Bases of Inherited Disease
prepared yeast or bacterial cells, respectively. The systems are set up such that the only cells that grow are those containing the appropriate yeastCitation:(inValle
Beaudet
AL, Vogelstein
B, Kinzler KW,
Antonarakis
SE, Ballabio
A, Gibson
K, the
Mitchell
G. The
Online
and
Molecular
selectable markers
the D,
case
of YACs)
or antibiotic-resistance
gene
(in the case
of BACs).
Note that
resulting
YACs
andMetabolic
BACs are
linear
and
Bases
of
Inherited
Disease;
2014
Available
at:
http://mhmedical.com/
Accessed:
July
12,
2017
circular DNA molecules, respectively.
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