The S1（Spike）protein gene without membrane localization signal of infectious bronchitis virus (dS1）was amplified from infectious bronchitis virus(M41) by PCR and sub-cloned into pBACsurf-1. The fusion gene containing S1 and gp64 was then inserted into baculovirus transfer vector pFastBac^TM Dual to construct the recombinant plasmid pFastBac-gp64-dS1.Then,the plasmid was transformed into Escherichia coli DH10Bac competent cells to obtain the recombinant shuttle vector Bacmid-gp64-dS1. Extracted Bacmid-gp64-dS1 was transfected into Sf9 cells to produce the recombinant baculovirus BV-dS1 by using Lipofectamine^TM 2000. Indirect immunofluorescent assay indicated that the recombinant baculovirus BV-dS1 could express S1 protein on infected Sf9 cell membrane.