Biomedical Engineering Reference
In-Depth Information
Most commonly used vectors now have a number of unique restriction
sites for this purpose in a region called the polylinker or multiple cloning
site (MCS). The regions immediately adjacent to the MCS often contain
specialized sequences that are used for a variety of purposes, such as
providing primer sites for sequencing the inserts (the cloned, inserted
DNA), polymerase sites for synthesizing RNA probes from the cloned
DNA. These flanking sequences can also contain other modifications
that enable the cloned DNA to be expressed in eukaryotic cells, or aid
in the analysis of promoter or enhancer activity of the cloned DNA. They
may also include additional sequences that permit the construction of
fusion proteins (see below). Plasmids used for molecular biology are
generally identified with a small ā€œpā€ at the beginning of their name. Com-
mon plasmids include the pBR vectors (pBR322, pBR328, etc.), pUC
vectors (pUC18 and pUC19, for example). Vectors within a series can
differ in the complexity or orientation of the polylinker, or contain different
or additional antibiotic resistance genes. Figure 3 shows an agarose gel
Figure
3.
Agarose gel electrophoresis of a restriction digest of a plasmid
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