摘要 :目的 构建人白细胞介素18(IL-18)基因复制缺陷型腺病毒表达载体。 方法 以质粒pJWIL-18为模板，设计并合成上、下游引物，利用多聚酶链反应(PCR)扩增获得IL-18基因片段，酶切后克隆至pCA13质粒，构建重组质粒pCA13IL-18。鉴定正确后，将pCA13IL-18与含有腺病毒右臂的质粒pBHGE3共转染HEK293细胞，9～12天出现病毒空斑。提取重组腺病毒的DNA，进行PCR鉴定。大量扩增后，氯化铯梯度离心纯化腺病毒，测病毒滴度。 结果 酶切分析、序列测定、PCR验证表明，IL-18基因成功克隆到复制缺陷型腺病毒载体中。 结论 成功构建了表达人IL-18基因的复制缺陷性腺病毒载体。

    关键词 :白细胞介素18;腺病毒载体

    Construction and identification of replication-deficient recombinant adenovirus vector expressing IL-18gene

    MAO Li-jun，ZHENG Jun-nian，ZHENG Dian-bao，et al

    (Department of Urosurgery，Affiliated Hospital of Xuzhou Medical College，Xuzhou，Jiangsu221002，China)

    Abstract:Objective To construct the replication-deficient recombinant adenoviruses vector expressing IL-18gene.Methods To get human IL-18template DNA sequences，one pair of specific primers to human IL-18gene were designed and synthesized and used to amplify IL-18cDNA by polymerase chain reaction(PCR)using pJWIL-18as the template.The PCR product was digested with EcoRⅠand BamHⅠand inserted into plasmid pCA13to construct the recombinant plasmid pCA13IL-18 ......