Cloning,Expression and Polyclonal Antibody Preparation of Murine Serum Amyliod p Component

Objective To prepare mSAP polyclonal antibody and make it use widely in mSAP detection.Methods The DNA fragment of coding region of the mouse SAP gene was amplied by PCR,then cloned it in to the prokaryotic expressed vector,PET30a(+).The recombinant plasmid was transferred into E.coli BL21.Fusion protein His/mSAP was expressed after IPTG was induced and isolated and purified from inclusion bodies.New Zealand rabbits were immunized with the purify fusion protein.The antiserum was detected by ELISA and Western blot.Results the prokaryotic expressed PET30a(+)/mSAP vector was successfully constructed,fusion protein His/mSAP was expressed efficiently.The polyclonal antibody was raised in the rabbits.Conclusion The mSAP antiserum reveal high titer and specificity and maybe applicable in SAP function research.