Cloning and expression of recombinant Vespa affinis hyaluronidase (rVesA2) were successfully expressed in Escherichia coli system. The VesA2 gene was cloned into pET-17b and pET-32a cloning vectors with a molecular weight of 41.71 and 59.0 kDa, respectively. The recombinant plasmid of pET-17b was composed of 1.08 kDa his-tag at the N-terminal. The 17.14 kDa of fusion tag, thioredoxin tag, histidine tag, and S-tag, was found in pET-32a. The verified expression conditions of rVesA2 induced under the conditions of 0.1 mM IPTG at 37◦C for 4 hrs gave the highest quantity of protein expression. The colony PCR and sequencing analysis were used to verify the rVesA2. The positive clones were detected the hyaluronidase activity by a zymographic gel. Recombinant proteins from both cloning vectors were insoluble. However, the recombinant form pET-32a showed higher solubility than pET-17b after dissolving in a 4 M urea solution. This result suggests that the fusion tag increases protein solubility.

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