禽类血管活性肠肽与乙肝病毒核心抗原融合基因的构建

在制备以禽类血管活性肠肽（VIP）为基础的基因工程疫苗工作中，选择乙肝病毒核心抗原（HBcAg)作为载体来提高鹅VIP的免疫原性，首先将克隆于鹅VIP cDNA和HBc基因第1至435bp的序列片段先后插入到质粒pRSET A的BamH I/EcoR I和Nhe I/BamH I克隆位点之间，构建成VIP序列位于HBc序列之后的VIP融合基因的重组质粒pHBc-VIP,其次将HBc第1至225bp序列的扩增片段和HBc第244bp之间包括VIP的充列经扩增，EcoR I酶切，连接，再扩增的片段先后插入到质粒pBSKS /－的BamHPst和PstHindⅢ克隆位点，构建成VIP持入到HBc基因中间（HB cAg的第75和82位氨基酸之间）融合基因的重组质粒pVIP-HBc.

Construction of Recombinant Fusion Genes with Goose VIP and Hepatitis B Core Antigen cDNAs

In the approaches of constructing recombinant vaccines based on avian vasoactive intestinal peptide (VIP), human hepatitis B core antigen (HBcAg) cDNA was chosen as the carrier for the purpose of enhancing immunogenicity of goose VIP. The VIP cDNA sequence and the 1 st to 435 th bp sequence of HBc cDNA (coding for 1 st to 145 th amino acid residues of HBcAg) were respectively amplified and inserted into BamH I Pst I and Nhe I BamH I cloning sites of plasmid pRSET A to produce plasmid pHBc VIP containing the HBc VIP fusion gene, which would express a fusion protein with VIP positioned posterior to the 145 th amino acid residue of HBcAg. Then the 1 st to 225 th bp sequence of HBc cDNA amplified, and the sequence starting from 244 th bp of HBc cDNA to the end of VIP sequence of pHBc VIP was amplified, digested with EcoR I, ligated and then amplified were respectively inserted into the BamH I Pst I and the Pst I Hind III cloning sites of plasmid pBSKS +/- to construct plasmid pVIP HBc, which contains a fusion gene with VIP sequence inserted into the middle of HBc cDNA, and coding for a fusion protein with VIP sequence inserted into the 75 th and 83 rd amino acid residue positions of HBcAg.