Construction of pET-32 α (+) vector for protein expression and purification
Ping-Chang Yang, Zhi-Qiang Liu
Abstract
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Ping-Chang Yang, Zhi-Qiang Liu
Abstract
Open-access reader
The construction of expression vector is a basic tool for biotechnology and production of desired proteins, this article summarized the construction of pET-32 α (+) 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 α (+) expression vector, protein expression in Escherichia coli BL21 (DE3) and protein purification under native conditions in E. coli lysates.
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The construction of expression vector is a basic tool for biotechnology and production of desired proteins, this article summarized the construction of pET-32 α (+) 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 α (+) expression vector, protein expression in Escherichia coli BL21 (DE3) and protein purification under native conditions in E. coli lysates.
Key concepts: Subcloning, Vector (molecular biology), Protein expression, Expression vector, Escherichia coli, Computational biology, Molecular biology, Target protein