Geneious logo

Virtual Cloning Tutorial: making a GFP fusion protein

This tutorial was developed by:
Dr Mike Bunce (Murdoch University, Perth, Western Australia) and the Biomatters team

In 2008 Osamu Shimomura, Martin Chalfie and Roger Y. Tsien, won a Nobel Prize in Chemistry for the discovery of Green Fluorescent Protein, GFP. Recombinant DNA technology (cloning) allows researchers to "attach" genes to this fluorescent marker and enables visualisation of where proteins localise within a cell.

In this tutorial, we will make a functional GFP fusion protein. The process of cloning involves an insert and a vector. Our insert is the Human Proinsulin gene and our vector is a plasmid named pEF-F3-EGFP which encodes an enhanced copy of the GFP gene (EGFP). Using the cloning tools in Geneious, we can conduct an in-silico experiment to investigate how best to make a plasmid with a functional fusion protein.

If you want to open the tutorial in a different window, you can click the new window button .


Introduction: Introduction to Green Fluorescent protein
Exercise 1: Cloning Vector
Exercise 2: Human Proinsulin
Exercise 3: Cloning Primer Design
Exercise 4: Introduce Cut Site using PCR
Exercise 5: Insert into Vector