Expression,purification and preservation of myofibrillogenesis regulator 1S recombinant protein and its antiserum preparation

Objective To obtain the myofibrillogenesis regulator 1S(MR-1S)-His recombinant protein through the expression of MR-1S gene and prepare anti-MR-1S antiserum after purification.To identify the stable system to preserve the recombinant protein,and to provid the reference for the investigation of MR-1S protein characteristics and biological function.Methods The recombinant plasmid pET21a-MR-1S was constructed with pronucleus expression vector pET 21a(+) and transferred into Escherichia coli BL21(DE3),and the expression of MR-1S recombinant protein was induced by isopropyl-beta-D-thiogalactopyranoside(IPTG).The MR-1S recombinant protein was purified by affinity chromatography and identified by Western blot.The anti-MR-1S polyclonal antibodies were prepared by the rabbit-immunized technique.With comparison of the protein activities of 4 stable systems under different temperatures,the most suitable stable preservation system and condition were picked out.Results Recombinant plasmid pET21a-MR-1S was established,and the MR-1S-His recombinant protein was expressed solubly in Escherichia coli BL21(DE3) by the identification of Western blot.The anti-MR-1S antisera were obtained by immunization rabbit,and the satabilization system and preservation condition were definited.Conclusions Under undenatured conditions,the soluble MR-1S recombinant protein is expressed and can be preserved stably,and it lays the foundation for MR-1S characteristices and biological function studies.