High‐efficiency expression of rat protein kinase C‐γ in baculovirus‐infected insect cells

Abstract
We have expressed rat protein kinase C‐γ in insect cells using a baculovirus vector. The yield of expressed protein kinase C‐γ is about 4% of total protein. The recombinant protein shows a prominent band at about 80 kDa on SDS‐polyacrylamide gels, which can be identified as protein kinase C‐γ by Western blotting using monoclonal antibodies against protein kinase C‐γ. Upon incubation with [γ‐32P]ATP and in the presence of Ca2+ phosphatidylserine and diacylglycerol this protein autophosphorylates. Its enzyme activity shows the characteristic properties of mammalian protein kinase C.