Shu L M, Horwitz M S, Engler J A
Department of Biochemistry, University of Alabama at Birmingham 35294.
Virology. 1987 Dec;161(2):520-6. doi: 10.1016/0042-6822(87)90146-2.
Replication of human adenovirus (Ad) DNA requires three virus-encoded proteins that are coordinately transcribed from a single promoter at early times after infection. The mRNAs for two of these proteins, the preterminal protein (pTP) and the Ad DNA polymerase (Ad Pol), share several exons, including one encoded near Ad genome coordinate 39. Plasmids containing the putative exons that encode Ad Pol mRNA were constructed to determine if enzymatically active Ad Pol protein could be synthesized. An Ad Pol of 140 kDa was detected by immunoprecipitation with specific antibody and its enzymatic activity was confirmed by complementation of Ad DNA replication in vitro. In addition to an Ad2 DNA fragment from 24.7 to 9.2 map units which contains an open reading frame for a protein of 120 kDa, the HindIII-J fragment that encodes the exon at genome coordinate 39 can be shown to be essential for production of full-length (140 kDa), enzymatically active Ad Pol.
人腺病毒（Ad）DNA的复制需要三种病毒编码蛋白，这些蛋白在感染后的早期从单个启动子协同转录。其中两种蛋白，即末端前蛋白（pTP）和腺病毒DNA聚合酶（Ad Pol）的mRNA共享几个外显子，包括一个在腺病毒基因组坐标39附近编码的外显子。构建了含有推定的编码Ad Pol mRNA外显子的质粒，以确定是否能合成具有酶活性的Ad Pol蛋白。用特异性抗体通过免疫沉淀检测到140 kDa的Ad Pol，其酶活性通过体外Ad DNA复制的互补作用得到证实。除了包含一个120 kDa蛋白开放阅读框的24.7至9.2图谱单位的Ad2 DNA片段外，编码基因组坐标39处外显子的HindIII-J片段对于全长（140 kDa）、具有酶活性的Ad Pol的产生至关重要。
