BE.109:DNA engineering

BE.109 Laboratory Fundamentals of Biological Engineering

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DNA engineering Protein engineering Systems engineering Bio-material engineering

Module 1

Instructors: Bevin Engelward and Natalie Kuldell

In this experimental module you will modify the gene for EGFP (Enhanced Green Fluorescent Protein) to truncate the protein it encodes. Cells expressing the full-length protein glow green when exposed to light of the appropriate wavelength. You will be designing and then creating an expression vector to delete the first 32 amino acids of EGFP. Cells transfected with your expression vector should not glow green, a prediction you will test. You will also test whether this N-terminally truncated EGFP can recombine with a C-terminally truncated version to regenerate full length EGFP in vivo. Finally you will examine the frequency of recombination when the C-terminally truncated version is “damaged” with different double-strand breaks.