Expression,purification,and identification of chloramphenicol acetyltransferase fusion protein

Objective To express and purify chloramphenicol acetyltransferase (CAT) gene. Methods The gene sequence which codes natural CAT portion was amplified with polymerase chain reaction (PCR). The fragment was cloned into prokaryotic expression vector pGEX-2T. The recombinant protein expression was induced by IPTG and the products were purified by agarose gel affinity chromatography resin. The antigenicity of purified recombinant CAT was identified by western blot. Results The recombinant CAT with a molecular weight of 52.6 kDAa was obtained in GST expression system. SDS-PAGE assay showed that the purified and digested product has high purity. The purified protein had the ability to reactive specifically with anti-CAT antibody. Conclusion The CAT fusion protein is expressed and purified, which lay a basis for preparation of antiserum and antibody.