Cloning and transformation of human multidrug resistance gene 1 (MDR1) / Hamidah Mokhtaruddin

Human MDR1 encoded for P-glycoprotein (P-gp) formation. P-gp is an ATP dependent efflux transporter which reduces the intracellular concentration of wide range of drugs including the chemotherapy drugs especially when the MDR1 gene being over expressed. Most of the anticancer agents are the substrates of P-gp thus make the tumour cells are able to survive upon exposure to various anticancer and this will lead to chemotherapy resistance. In order to study about the mechanism of P-gp, human MDR1 has been cloned and transformed into plasmid of E.coli as the reaction vector. Before cloning and transformation take place, amplification of MDR1 gene was done using Polymerase Chain Reaction (PCR) and followed by gel electrophoresis. The appeared band on gel with 158 bp showed present of desired gene, thus progressed on cloning and transformation methods. As colonies obtained on the selective agar, positive transformants analysis is followed by doing the second PCR and sequencing. These procedures are to confirm the MDR1 gene is cloned at the correct orientation.