Purification of CT3 Fragment of Cav1.2 with Dithiothreitol / 中国医科大学学报

ABSTRACT

Objective To explore whether dithiothreitol(DTT)is helpful for PreScission Protease to cut off the GST from GST?CT3 protein. Meth?ods The pGEX?6P?3/CT3 recombinant plasmid was transfected into Escherichia coli BL21,and the GST?CT3 fusion protein was purified by B?PER method. PreScission Protease was applied with 10 mmol/L DTT to cut off the GST,then the SDS?PAGE was performed for identification of the CT3 protein. Results Without DTT,it was very difficult for PreScission Protease to cut off the GST from GST?CT3 protein. However,in the pres?ence of 10 mmol/L DTT,PreScission Protease could cut off the GST easily as identified by SDS?PAGE. Conclusion 10 mmol/L DTT can help Pre?Scission Protease to cut GST from GST?CT3 protein,so as to achieve high concentration of CT3.