Objective Two - Isolate and Purify Reflectin in vitro

Purification of recombinant reflectin from bacteria was necessary in order to perform all of the subsequent in vitro studies of the protein. An over-exressing vector construct was used to maximise yield, and a his-tag was used for purification purposes.

Tagging the reflectin gene

The reflectin genes were his-tagged by incorporating the his sequence into the primers used to clone the gene. His tagging was chosen because...

Overexpression

We chose to use a high copy plasmid with a pBAD promotor in order to overexpress reflectin. This produces inclusion bodies, which will create unfolded reflectin, but this is not a concern because the solvents

Protein extraction and purification