TABLE OF CONTENTS INTRODUCTION
This set of experiments is organized as a set of interconnected processes. The initial exercises in Part 1 (Lab 1.1) involve growing E. coli cells and subsequent isolation of plasmid DNA containing the myo-3 gene fragment from the nematode C. elegans. During the same lab, in a parallel procedure, the plasmid DNA that will receive the myo-3 fragment (via ligation) and then be used for protein expression is isolated. The plasmid strands are cleaved with a sequence-specific restriction endonuclease ( HindIII), then the desired fragment is resolved from the remaining DNA by electrophoresis on a polyacrylamide gel (Lab 1.2). The amount of DNA fragment is quantitated, then to obtain pure DNA fragment the band is excised from the gel (Lab 2.1). In Lab 2.2, the myo-3 fragment is ligated into the expression plasmid, which is equipped with sequences that direct transcription of the appropriate messenger (m)RNA. This mRNA is capable of being bound by the ribosome and decoded to make myo-3 linked to the protein ?-galactosidase, which comes connected to the fusion protein expression vector. In Lab 3.1, a DNA fragment is produced in large quantities from the...
