禽类血管活性肠肽与乙肝病毒核心抗原融合基因的构建  

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

作　　者：施振旦[1] 黄运茂[1] 曹永长[1] 毕英佐[1] 

机构地区：[1]华南农业大学动物科学系,广东广州510642

出　　处：《华南农业大学学报》2001年第3期60-63,共4页Journal of South China Agricultural University

基　　金：国家自然科学基金资助项目 (3970 0 10 2 )

摘　　要：: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VIPcDNA和HBc基因第 1至 435bp的序列片段先后插入到质粒pRSETA的BamHI\EcoRI和NheI\BamHI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 2 2 5bp序列的扩增片段和HBc第 2 44bp之后包括VIP的序列经扩增、EcoRI酶切、连接、再扩增的片段先后插入到质粒pBSKS + / -的BamHI\PstI和PstI\HindIII克隆位点 ,构建成VIP插入到HBc基因中间 (HBcAg的第 75和 82位氨基酸之间 )融合基因的重组质粒pVIP HBc .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.

关 键 词：血管活性肠肽 乙肝病毒核心抗原 融合基因 禽类 

分 类 号：S858.364[农业科学—临床兽医学] S859.794[农业科学—兽医学]