Prokaryotic Expression and Purification of Actin Depolymerizing Factor in Rubber Tree
In this study, the primer pair was designed to amplify the open reading fragment of HbADF according to the known HbADF sequences. The amplified fragment was inserted into pET28a vector to construct the recombinant vector of pET28a-HbADF. The recombinant vector was transformed into an Escherichia coli strain BL21(DE3), and the HbADF protein with molecular weight about 20 ku was successfully induced by adding 1mM isopropyl-β-D-thiogalactopyranoside(IPTG). The fusion HbADF protein was produced in the form of the solubility or inclusion body, purified by affinity chromatography, identified by Western blot with anti-HIS tagged monoclonal antibody. These results laid a foundation for elucidating characteristic and function of HbADF in rubber tree.