中文摘要：

目的构建含丙型肝炎病毒（HCV）核糖体插入位点（IRES）序列的基因克隆,为以后的亚克隆和抑制肝炎病毒作用的研究提供实验材料。方法以含HCV全长基因的质粒为模板,用PCR技术扩增出HCV的IRES序列,将扩增产物IRES基因插入到PMD18T载体后转化DH5α,筛选阳性克隆,抽提重组质粒并进行PCR及酶切鉴定,再行序列分析。结果经PCR获得355bp含限制性内切酶位点的阳性产物,T载体克隆、PCR及酶切鉴定和序列分析后证实,克隆片段与GeneBank中该基因的序列同源性为99%。结论该实验成功构建了含HCV IRES基因序列的T载体克隆,提示该克隆是用作亚克隆和抑制肝炎病毒研究的理想克隆。

英文摘要：

Objective To construct the PMD18T vector containing the internal ribosomes of entry site （IRES） gene of the hepatitis C virus （HCV） and provide experimental material for subelone and the study on the inhibition of hepatitis virus. Methods The IRES gene was amplified by PCR from the plasmid containing the complete genome of HCV. The amplified DNA fragments were ligated into PMD18T vector,and then transformed into DH5α. The positive clones were screened out and the recombinant plasmids were isolated from such clones. The recombinants were identified by PCR, restriction endonueleases analysis and sequencing. Results PCR assay showed that a DNA fragment of 355bp could amplified from the recombinant plasmid. Restriction endonucleases digestion confirmed the target fragment had been inserted into PMDI8T successfully. The sequence results verified that the sequence of the target fragment had high homology （ 99% ） with the IRES gene sequence of HCV in GeneBank. Conclusion The T vector clone with IRES gene of HCV has been constructed successfully and it is effective for the study on subcloning and the inhibition of hepatitis virus.