2. Cell Cycle

3. Cell Cycle Control of DNA Replication
Initiation of replication involves replicators and the
origin recognition complex, or ORC, a heteromeric
protein that binds to replicators. ORC is bound to
the replicators throughout the cell cycle. Early in G1
(just after M), ORC serves as a “landing pad” for
proteins essential to replication control. Proteins binding
to ORC establish a pre-replication complex (pre-RC),
but the pre-RC can be formed only during a window
of opportunity during G1. One of the principal proteins
in assembly of the pre-RC in yeast is Cdc6p (the
replication activator protein encoded by the yeast
cdc6 gene). Once Cdc6p binds to ORC, replication
licensing factors (RLFs) that “license” or permit
DNA replication to occur then bind to the
chromosomes.

6. Fluorescence Activated Cell Sorting (FACS)

7. FLAG-Tag Plasmid (Example)
HA-Tag: (hemagglutinin)
YPYDVPDYA

8. PCNA (proliferating cell nuclear antigen)
The protein encoded by this gene is found in the nucleus and is a cofactor of
DNA polymerase. The encoded protein acts as a homotrimer and helps
increase the processivity during DNA replication.

14. DpnI cleaves DNA only when it is methylated on both strands.
And mammalian cells lack the ability to methyate Dpn I restriction sites.