Amplification of human pregnane x receptor gene by using polymerase chain reaction / Roziwaty Anzara Azman

Polymerase chain reaction (PCR) is a fast and inexpensive technique used to amplify, or make many copies of, small segments of DNA. In the mean time, cloning is another method to amplify a specific gene that involves transferring a DNA fragment of interest from one organism to a self-replicating genetic element such as a bacterial plasmid ( cloning vector). This followed by propagation of DNA of interest in a foreign host cell. In this study, the objective of this study is to amplify Human Pregnane X Receptor (hPXR) Gene using Polymerase Chain Reaction for use in cloning and expression. Initially, both forward and reverse primers for hP XR gene are designed followed by reconstitution of the primers to prepare working stock for the polymerase chain reaction (PCR). The composition for PCR experiment was determined and a suitable PCR condition was designed to amplify the primers. Gel electrophoresis then was performed and the bands appeared were visualized under UV transilluminator. The result showed that the product of hPXR gene was obtained by using 55 °c as annealing temperature. As a conclusion, in this study the hPXR gene was successfully amplified. This product could be used further for the expression of the protein.