The DNA fragments coding for preS2(120 146) and preS1(21 47) amplified by PCR were fused to both 5′ and 3′ ends of S gene at the position of amino acid 223. The fusion gene was placed downstream of the promoter P7.5...The DNA fragments coding for preS2(120 146) and preS1(21 47) amplified by PCR were fused to both 5′ and 3′ ends of S gene at the position of amino acid 223. The fusion gene was placed downstream of the promoter P7.5 of the universal vaccinia viral vector pGJP 5 and the recombinant vaccinia virus vS2SS1 was then selected by \%in vivo\% homogeneous recombination. Fusion protein S2SS1 could be expressed in the mammalian cells infected with vS2SS1. The investigation of expression, secretion, antigenicity and particle assembly of the S2SS1 protein demonstrated that S2SS1 protein could be assembled into particles which presented preS1, preS2 and S antigenicity and be efficiently secreted from the cells. It also showed that the level of its expression and secretion approached to that of the S protein expressed by the recombinant vaccinia virus.展开更多
文摘The DNA fragments coding for preS2(120 146) and preS1(21 47) amplified by PCR were fused to both 5′ and 3′ ends of S gene at the position of amino acid 223. The fusion gene was placed downstream of the promoter P7.5 of the universal vaccinia viral vector pGJP 5 and the recombinant vaccinia virus vS2SS1 was then selected by \%in vivo\% homogeneous recombination. Fusion protein S2SS1 could be expressed in the mammalian cells infected with vS2SS1. The investigation of expression, secretion, antigenicity and particle assembly of the S2SS1 protein demonstrated that S2SS1 protein could be assembled into particles which presented preS1, preS2 and S antigenicity and be efficiently secreted from the cells. It also showed that the level of its expression and secretion approached to that of the S protein expressed by the recombinant vaccinia virus.
