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CHEM 3525
Course Title:
Biochemistry II Lab
This course will be an elective for the Minor in Chemistry, and will eventually be an elective or
required upper-level division course for the Major in Chemistry as that is developed. As we
already offer the 2000-level year-long series of Organic Chemistry, there will be a number of
students well-prepared to take this class to complete requirements for the Minor and Major in
Chemistry.
Upon completion of the course, each student should:
 Know how to use literature and information search engines.
 Understand how to design primers for cloning a gene of interest into a recombinant DNA
vector.
 Be able to create mutants through site–directed mutagenesis.
 Learn about sequencing.
 Be trained how to induce overexpression of proteins into E.Coli.
 Know how to purify proteins through Nickel Chromatography.
 Be able to characterize them by Coomassie staining.
This is a hands-on laboratory course where students will develop their own biochemical research
project. They will learn how to clone, express and purify a protein of interest. A Histidine tagged
version of wild-type GFP will be engineered. Students will then design GFP mutants by sitedirected mutagenesis. They will choose their own GFP mutations by studying the literature. Once
the mutants are confirmed by sequencing, proteins will be overexpressed into E.Coli and purified
by Nickel Chromatography. Coomassie staining will confirm the presence of wild-type and
mutant GFP proteins.
Assessment will include individual and group assignments (25% of total grade), exams (25% of
total grade), a final exam (20% of total grade), participation in class discussions (10%), and a
report/proposal based current topics and reports in the scientific literature (20%).
Biochemistry
Jeremy M. Berg, John Tymoczko, Lubert Stryer
@2012 Seventh Edition
ISBN-13: 9781429229364
Other Information:
Areas in bold are required.