Prokaryotic Expression and Polyclonal Antibody for Ferric Uptake Regulator Prepared from Vibrio vulnificus FJ03-X2

Ferric uptake regulator (fur) gene from Vibrio vulnificus FJ03-X2 was cloned, and inserted into the prokaryotic expression vector, pET32a. Coding sequence of the gene contained 450 bps, encoding 149 amino acids. Its sequencing showed a greater than 98% homology with that of the fur gene from GenBank. Subsequently, the plasmid, pET32a-FUR, was transformed into E. coli BL21 to express the recombinant protein, rFUR. The soluble rFUR was then subject to the Ni-NTA His Binding affinity purification. The purified rFUR was injected into SD rats to produce polyclonal antibody. The obtained highly specific antibodies of FUR was confirmed by ELISA assay.