【摘要】 目的 获得7型腺病毒(Ad7)载体构建的基本元件，为构建Ad7载体及阐明其基因组一级结构打下基础。方法 用改良Hirts法提取Ad7 DNA，酶切后克隆于质粒载体，克隆了Xho Ⅰ A+D片段(17.0-76.5mu)、D片段(17.0-22.9mu)、E片段(10.8-14.1mu)、F+G片段(14.1-17.0mu)、G片段(15.8-17.0)，用EcoR Ⅰ+Xho Ⅰ双酶切，克隆了76.5-87mu，用Sam Ⅰ+EcoR Ⅰ双酶切，克隆了87.0-97.4mu。用腺病毒DNA做为模板测定了Ad7疫苗株右侧末端Sma Ⅰ H片段(97.4-100mu)核酸苷序列，根据所测序列设计引物，用聚合酶链反应(PCR)获得右侧末端，并对其再次测序证实。结果 建立了Ad7疫苗株部分基因文库(10.8-100mu)，完成Ad7疫苗株Sma Ⅰ H片段核苷酸序列。结论 这一结果为阐明Ad7基因组一级结构及构建Ad7载体奠定了基础。

Abstract Human adenoviruses have been a hot topic of study because of their potential utility for gene therapy and development of live viral vectored vaccines. Adenovirus types 4 and 7 have been given orally as vaccines for prevention of acute respiratory infections caused by these serotypes of adenovirus in U.S. army recruits for more than 30 years. Therefore, the advantage of using Ad4 and Ad7 as vectors in the development of oral live recombinant vaccine is obvious. For constructing Ad7 vector, 10.8-100mu DNA of Ad7 vaccine strain has been cloned. The Sma Ⅰ H fragment was sequenced using Ad7 DNA as a template and then the fragment was cloned by PCR. For verifying the sequence the fragment was sequenced again. The strategy we used to clone terminal end of adenovirus is reliable and practicable.

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