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Background  Pigeon circovirus (PiCV) is considered to be a viral agent central to the development of young pigeon disease syndrome (YPDS). The Cap protein, a structural protein encoded by the  cap  (or C1) gene of PiCV, has been shown to be responsible for not only capsid assembly, but also has been used as antigen for detecting antibody when the host is infected with PiCV. The antigenic characteristics of the Cap protein potentially may allow the development of a detection kit that could be applied to control PiCV infection. However, poor expression and poor protein solubility have hampered the production of recombinant Cap protein in the bacteria. This study was undertaken to develop the optimal expression of recombinant full-length Cap protein of PiCV using an  E. coli  expression system.    Results  The PiCV  cap  gene was cloned and fused with different fusion partners including a His-tag, a GST-tag (glutathioine-S-transferase tag) and a Trx-His-tag (thioredoxin-His tag). The resulting constructs were then expressed after transformation into a number of different  E. coli  strains; these then had their protein expression evaluated. The expression of the recombinant Cap protein in  E. coli  was significantly increased when Cap protein was fused with either a GST-tag or a Trx-His tag rather than a His-tag. After various rare amino acid codons presented in the Cap protein were optimized to give the sequence rCap opt , the expression level of the GST - rCap opt  in  E. coli  BL21(DE3) was further increased to a significant degree. The highest protein expression level of GST - rCap opt  obtained was 394.27 ± 26.1 mg/L per liter using the  E. coli  strain BL21(DE3)-pLysS. Moreover, approximately 74.5% of the expressed GST - rCap opt  was in soluble form, which is higher than the soluble Trx-His-rCap opt  expressed using the BL21(DE3)-pLysS strain. After purification using a GST affinity column combined with ion-exchange chromatography, the purified recombinant GST - rCap opt  protein was found to have good antigenic activity when tested against PiCV-infected pigeon sera.    Conclusions  These findings shows that the  E. coli -expressed full-length PiCV Cap protein has great potential in terms of large-scaled production and this should allow in the future the development of a serodiagnostic kit that is able to clinically detect PiCV infection in pigeons.

High yield production of pigeon circovirus capsid protein in the E. coliby evaluating the key parameters needed for protein expression