Abstract：Objective To clone and construct a BL21 expressing strain containing insulin related polypeptide 3 (IRP3) gene, and isolate, purifiy, identifiy and renaturat the expressed protein. Methods IRP3-pET30a positive strain BL21 was cloned by DNA recombination technique, the plasmid IRP3-PET30A was successfully constructed by agarose gel electrophoresis after double enzyme digestion, and IRP3/HIS purified protein was obtained by nickel affinity chromatography, SDS-PAGE and Western blot were further used to confirm the purification effect of IRP3/HIS fusion protein. Results Plasmid IRP3-PET30A was successfully cloned and BL21 expressing strain containing plasmid IRP3-pET30a was screened, purified IRP3/HIS fusion protein was obtained from the supernatant of inclusion body lysis and renaturated by dialysis. Conclusion The successful cloning of IRP3 gene and the expression and purification of IRP3/HIS fusion protein lay a foundation for the preparation of IRP3 protein antibody and the application of IRP3 protein as in vitro growth factor additive. What we have done will surely be useful to the antibody preparation for the IRP3 protein and the application of growth factor additive in vitro.
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