Cloning and expression of phosphoenolpyruvate carboxykinase from Escherichia coli

Abstract

In order to better study the structure and function of phosphoenolpyruvate carboxylase, the phosphoenolpyruvate carboxylase gene is cloned and expressed by PCR (polymerase chain reaction), double enzyme digestion and cell transformation. The results show that the new phosphoenolpyruvate carboxylase gene of Escherichia coli is successfully amplified and cloned into the prokaryotic expression vector pET-30a, and then introduced into Escherichia coli BL21 expression system. The phosphoenolpyruvate carboxylase of Escherichia coli is successfully induced by IPTG. The phosphoenolpyruvate carboxylase can be efficiently expressed by using the proposed method, which provides preparation for further scale expression, purification and application.

Keywords

• genetic engineering
• Escherichia coli
• phosphoenolpyruvate carboxykinase
• protein expression
• CO2