Construction of pET-32 a (+) Vector for Protein Expression and Purification

The construction of expression vector is a basic tool for biotechnology and production of desired proteins, this article summarized the construction of pET-32 alpha (+) vector techniques which are generally used in research laboratories. The procedures include that acquisition of the exogenous DNA fragment for construction of the vector, subcloning the DNA fragment into pET-32 alpha (+) expression vector, protein expression in Escherichia coli BL21 (DE3) and protein purification under native conditions in E. coli lysates.