Recombinant DNA MethodMutagenesis
By: Jessica Green
Mutagenesis
Mutagenesis (the creation or
formation of a mutation) can be used
as a powerful genetic tool. By
inducing mutations in specific ways
and then observing the phenotype of
the organism the function of genes
and even individual nucleotides can
be determined.
Mutagenesis Laboratory
Some types of Mutagenesis
Directed Mutagenesis
Site-directed/Site-specific
Mutagenesis
Mismatched Mutagenesis
Directed Mutagenesis
a largely discredited hypothesis
proposing that organisms can
respond to environmental stresses
through directing mutations to certain
genes or areas of the genome.
Site-directed Mutagenesis
Where a specific site in a cloned DNA
needs to be altered in a precise, predetermined way
Can be designed to create specific
nucleotide substitutions, deletions,
and so on
Image of Site-directed Mutagenesis
Two different forms of Site-specific
Mutagenesis
5’ add-on mutagenesis
- Diagram on page 148 of the text
- a new sequence or chemical group
is added to the 5’ end of a PCR
product
Mismatched primer mutagenesis
- primer is designed to be only
partially complementary
Mismatched Mutagenesis
Diagram pg. 147 of text; Figure 5.19
Can create a desired point mutation
at a unique predetermined site within
a cloned DNA molecule
At the intended mutation site it bears
a base that is complementary
References
http://humogef.ifj.edu.pl/zaklad/laborat
or.html (pictures)
Textbook
http://www3.interscience.wiley.com/cg
ibin/jhome/10009058?CRETRY=1&S
RETRY=0