Preparation of a specific CYP2C19 transcript for in vitro cloning / Shafinaz Ahmad

Cytochrome P450 2C 19 is a member of cytochrome P450 that is involve in the metabolism of several important groups of drugs including many proton pumps inhibitor and antiepileptic. Genetic polymorphisms of CYP2Cl9 are associated with variable ability to metabolize mephenytoin, known as the poor metabolizer and extensive metabolizer phenotypes. The aim of this study is to prepare human CYP2Cl9 cDNA for cloning in E. coli. Temperature gradient PCR method was used to amplify the CYP2C. A set of primers have been designed to produce the target sequence. Few parameters were frequently adjusted to optimize the PCR reaction for higher specificity and reproducibility. CYP2Cl9 had to be isolated from the other CYP2C genes by using restriction enzyme because of their high similarity in gene sequences. At the end of the study, the result was successful. For the conformation of the product that had been produced, the sequencing process should be done for further study.