Prokaryotic expression and serum preparation of envelope protein VP53B of white spot syndrome virus (WSSV)

Abstract:

[Objective] To clone, express and purify envelope protein sVP53B of WSSV (white spot syndrome virus) for polyclonal serum preparation. [Methods] Based on the GenBank sequence of WSSV vp53B, primers were designed, and PCR was done to amplify functional sequence of Svp53B. Svp53B was inserted into prokaryotic expression vector, pET-16b. Recombinant plasmid was transformed into Escherichia coli Rosetta 2. The expression of sVP53B was detected by SDS-PAGE and Western blotting. The protein was purified with Ni-NTA agarose beads, then applied to immune rabbits for polyclonal serum detecting it’s titer by indirect ELISA (enzyme-linked immuno sorbent assay). [Results] The sVP53B was expressed most by 1 m mol/L IPTG (isopropyl-β-D-thiogalactoside) inducing and mainly expressed as inclusion bodies. Polyclonal serum, prepared by immunizing rabbit, revealed a titer as high as 1:150 000. [Conclusion] Recombinant protein sVP53B was highly expressed in E. coli and showed high purity after purification. The prepared polyclonal serum had high affinity and specificity against sVP53B. The results pave the way for functional study of VP53B during WSSV oral infection.