Preparation of feline herpesvirus type 1 VLPs in yeast and assembled in vitro
To develop virus-like particles(VLPs)vaccines against feline herpesvirus type 1(FHV-1),FHV-1 gD gene was fused with self-assembling protein 1301 gene and then cloned into pPink-HC vector.The recombinant gD protein was expressed in Pichia pastoris.The effects of different pH on protein expres-sion and VLPs assembly were then observed.The results showed that P.pastoris expressed intact recombi-nant gD protein at pH3.0,which was identified by SDS-PAGE and Western-blot,while the molecular weight of protein expressed under pH6.0 condition was smaller than expected.In addition,the assembly effect of VLPs at pH8.0 was better than that of pH6.0 and 7.0.These results showed that acidic environment could effectively inhibit the hydrolysis of recombinant gD protein by proteases in P.pastoris.Recom-binant gD protein can be assembled into VLPs in vi tro by using self-assembled protein 1301 in a weak al-kaline environment,which lays a foundation for preparation of FHV-1 VLPs vaccine.
feline herpesvirus type 1virus-like particlesyeast expression systemgD gene
国家科技期刊平台
NSTL
万方数据
