ISSN: 0973-7510
E-ISSN: 2581-690X
Deoxycytidine kinase (DCK) is required for the phosphorylation of several deoxyribonucleosides and their nucleoside analogs. Deficiency of DCK is associated with resistance to antineoplastic chemotherapeutic agents. We developed a baculovirus expression system to express the DCK protein directly in silkworm. A recombinant dual expression vector containing his-tagged dck and polyhedrin (polh) was constructed, and baculovirus polyhedra were generated by transfecting the recombinant plasmid into BmN cells and used to orally inoculate the silkworm larvae. The results showed that the His-tagged DCK fusion protein was successfully expressed in the silkworm larvae and purified by Ni – NTA affinity chromatography, and ELISA quantative assay showed that recombinant DCK protein enzymatic activity up to 106.79U/L. These results proved that it is feasible and convent expression system to express DCK protein in silkworm through oral feeding.
DCK, pFastBacDual, His-tag, Recombinant baculovirus, Bac-to-Bac system
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