Gene Therapy: Lab Report Assignment

The LIEU gene Is a linear fragment that does not notation an Autonomous Replication Sequence, so it could not replicate on its own and needed to be integrated by homologous recombination. The TRIP gene was a circular plasmid that contained an EARS, which allowed for it to act as an extra chromosome In the gene. The objective was to insert a “wild gene” and replace the defective genes and then grow them on a medium that does not contain TRIP or LIEU to prove that the genes had been cured.Gene Therapy: Lab Report Assignment

To help determine if recombination took place in the LIEU gene, and to compliment negative data from the 431 LIEU drop out medium, the “cured” LIEU gene was compared to the “diseased” ELISE gene. The expectation was that the “cured” LIEU gene would be a different size from that of the “diseased,” which would be proven through a PC run of the two DNA strands after they were replicated under the same in vitro conditions. The purpose of the PC was to show what kind of mutation occurred in the mutant to cause It to lose Its LIEU function.