Expression of Fusion Protein of hEGF and gp67 Signal Peptide in Larvae of Bombyx mori

Abstract:

Bombyx mori baculovirus expression vector system (BEVS) was adopted to express S-hEGF. The reconstructed plasmids of pBacPAKS-hEGF and an engineering modified BmNPV Bm-BacPAK6 DNA were used to co-transfect BmN cells in order to obtain recombinant virus. The recombinant virus (named vBacPAKS-hEGF) was then used to infect BmN cultured cells, the fifth instars larvae and pupa. The aimed 12 kD protein expressed in BmN cultured cells and the fifth instars larvae was identified with Western blot. ELISA results showed that the amount of expression protein reached 23 mg/ 106 in BmN cells and 82 mg/mL in larvae. The biological activity was determined by epidermal growth factor dependent Balb/c3T3 cell line. Both cellular extracts and haemolymph of silkworm larvae infected with the recombinant virus accelerated proliferation of Balb/c3T3 cells. In addition, animal experiment results revealed that S-hEGF fusion protein promoted newborn ICR mice weight increasing, incisor eruption and eyelid opening.