Expression of a functional recombinant oleosin-human hyaluronidase hPH-20 fusion in Arabidopsis thaliana

• The hyaluronidase PH20 protein was expressed in plants for the first time.
• The oleosin-fusion technology was first used to express hPH20 protein.
• The cell activity method of hPH20 protein expressed in plant oil body was established.

The use of plants as expression systems for the production of recombinant proteins has distinct advantages, such as safety, ease, low cost and high yields. A plant binary expression vector, pOTBar-hPH20, containing an oleosin-hPH20 fusion gene and a Basta selection marker gene was constructed and introduced into Arabidopsis thaliana via the floral dip method. Transformed A. thaliana seed lines were obtained and analyzed by PCR. The PCR results indicated that oleosin-hPH20 fusion gene was integrated into the A. thaliana genome. The oleosin-hPH20 fusion protein was detected by SDS–PAGE and Western blot analysis. The oleosin-hPH20 fusion protein was expressed and had good antigenicity in the transgenic A. thaliana seeds. An enzyme assay suggested that the recombinant oleosin-hPH20 fusion protein had hyaluronidase activity.