【摘要】 应用基因重组技术。从载体ＣＤＺ２酶切获得１．７３ｋｂＤＮＡ片段（含１６ｄ９３ｂｐ的ＨＣＶ结构区ｃＤＮＡ，其特异性已为ｓｏｕｔｈｅｒｎｂｌｏｔ证实）。将ＨＣＶｃＵＮＡ片段插入载体ｐｃＤＮＡ３，构建ＨＣＶ重组体。经在大肠杆菌中表达，筛选、酶切分析，获得重组ＨＣＶ（ＰｃＤＮＡ３－ＨＣＶ）。以重组质粒转染Ｈ９细胞（ＣＤ淋巴细胞系），用Ｇ４１８筛选出抗性细胞。经ＲＴ－ＰＣＲ、ｄｏｔ－ＥＬＩＳＡ验证表明，ｐｃＤＮＡ３－ＨＣＶ能在Ｈ９细胞中进行复制、转录和表达。这为丙型肝炎发病机理的研究提供了一个有用模型。更多还原

【Abstract】 By inserting 1693bp HCV cDNA (total structure region genes) from CDZ2 into pcCNA3 vector, the recombinant HCV expressive vector pcDNA3 - HCV was constructed. To identify the recombinant HCV , it was intoduced into JM109 E.coli strain , then restriction enzyme - reaction was carried out respectively. H9 cell line(CD T cell) was transfected by the recombinant HCV expressive vector pcDNA3 - HCV and G418 resistant cell clones were identified. RT - PCR, dot ELISA showed that the pcDNA3 - HCV was replicated. transcribed and expressed in H9 cells. The construction of the H9 cell model transfixed by pcD) -- HCV was successful.更多还原