Cloning, Expression and Purification of Human DNA Polymerase δ Interaction Protein PDIP38

[Objective] Clone, express and purify wild type and 3 deletion mutants of human PDIP38 glutathion S transferase (GST)fusion protein. [Methods] Polymerase chain reaction (PCR) amplification was used for cDNAs of wild type and deletion mutants of PDIP38. 4 cDNAs were inserted into the pGEX-4T-l GST fusion vector and transformed to E coli. DH5α cell strain. Restriction enzyme BamH I and Xho I were used for double digest to check the inserted DNA. Glutathione-sepharose 4B affinity column was used to purify PDIP38 recombinant proteins. [Results] GST fusion wild type and deletion mutants of PDIP38 were highly expressed in E coli. DH5α. The purities of the recombinant proteins achieve 85% on glutathione-sepharose 4B affinity column. [Conclusions] pGEX-4T-l GST fusion protein vector can highly express human PDIP38 proteins. Using Glutathione-sepharose 4B affinity column can simply and rapidly purify the proteins.