Know the proteins that are able to help DNA stay in a single stranded form

Understand the composition of the DNA polymerase III holoenzyme and how the different factors interact with each other

Understand how Okazaki fragments are connected together

The structure of oriC (origin of replication in E. coli)

Understand how replication initiation is controlled

Be able to describe the end replication problem and how it can be overcome

The ultimate goal: Be able to label and describe the figure on slides 15 and 16

For the following mechanisms, understand/describe (to the detail discussed in lecture):

• The sequence features required for the mechanism to take place
• The steps of the mechanism
• The proteins involved at each step
• The outcome(s) of the mechanism

Mechanisms:

Replication initiation

• DNA melting
• priming

Replication

• Coordination of replication on the leading and lagging strands
• Termination of replication

For mechanisms as well as the other proteins mentioned in the learning objectives, think about what would happen if you removed their functionality through mutagenesis.

Assays/techniques to be familiar with:

Temperature sensitive mutants Including the difference between quick stop and slow stop mutants