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From gene mutation to protein characterization
Author(s) -
Moffet David A.
Publication year - 2009
Publication title -
biochemistry and molecular biology education
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.34
H-Index - 39
eISSN - 1539-3429
pISSN - 1470-8175
DOI - 10.1002/bmb.20257
Subject(s) - mutant , green fluorescent protein , mutation , sequence (biology) , microbiology and biotechnology , point mutation , gene , biology , protein sequencing , sequence analysis , computational biology , genetics , peptide sequence , biochemistry
A seven‐week “gene to protein” laboratory sequence is described for an undergraduate biochemistry laboratory course. Student pairs were given the task of introducing a point mutation of their choosing into the well studied protein, enhanced green fluorescent protein (EGFP). After conducting literature searches, each student group chose the mutation they wanted to introduce into EGFP. Students designed their sequence‐specific mutagenic primers and constructed their desired mutation. The resulting EGFP mutant proteins were expressed in E. coli , purified and characterized. This laboratory sequence connected the major concepts of molecular biology and biochemistry, while incorporating the thrill of novel discovery in an undergraduate‐level biochemistry laboratory course.