Site directed mutagenesis of V2 vasopressin receptor and its cloning using PGEM3Z vector

Abstract

The present study was conducted to create mutations in this motif for further investigation of its function. By using nested PCR two mutations were produced, which would translate to DRY and DKH. The PCR products as well as the PGEM3Z vector were digested using NooI and EcoRI restriction enzymes and were ligated and transformed to E. coli HB101 cells. The obtained colonies were analyzed for the presence of the inserts using suitable restriction enzymes. The obtained plasmids have the advantage of having restriction sites, which would not interfere with further cloning and expression of this receptor using mammalian expression vectors. © 2005 Asian Network for Scientific Information.