5.4 Site-Directed Mutagenesis
In site-directed mutagenesis, the enzyme is used to synthesize the mutant DNA strand from a single-stranded DNA template.
A specific , such as NciI, is used to selectively nick the DNA template strand while leaving the newly synthesized mutant strand intact.
Next, is used to progressively digest the 3’ end of the nicked DNA template strand.
The gap created by exonuclease III is filled in using the enzyme and the DNA backbone is resealed by .
True or False: In site-directed mutagenesis, the DNA template must be linear, double-stranded DNA.
A. |
B. |
True or False: In site-directed mutagenesis, the oligonucleotides can be designed to introduce point mutations, insertions, or deletions into the gene sequences.
A. |
B. |
True or False: M13 bacteriophage DNA is often used in site-directed mutagenesis experiments as the template strand because it is produced as a circular, single-stranded DNA molecule.
A. |
B. |
In the example shown, why is it important that the cytosine residue incorporated into the newly synthesized mutant DNA contains a sulfur atom substituted for one of the oxygen atoms?
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