Sarnow P
Department of Biochemistry, Biophysics and Genetics, University of Colorado Health Sciences Center, Denver 80262.
J Virol. 1989 Jan;63(1):467-70. doi: 10.1128/JVI.63.1.467-470.1989.
It has been shown by van der Werf et al. (S. van der Werf, J. Bradley, E. Wimmer, F. W. Studier, and J. Dunn, Proc. Natl. Acad. Sci. USA 83:2330-2334, 1986) that in vitro synthesis of poliovirus RNA by T7 RNA polymerase gives rise to infectious RNA molecules; however, these molecules are only 5% as infectious as RNA isolated from virions. A plasmid, T7D-polio, was constructed that allows the in vitro synthesis of full-length RNA molecules with two additional guanine residues at the 5' end. However, T7D-polio differed from the construct of van der Werf et al. in that RNA transcribed from T7D-polio has an authentic 3' end, ending with only a polyadenine nucleotide sequence. Transfection of these RNA molecules into mammalian cells produced wild-type poliovirus with an efficiency similar to that of virion RNA. The use of this vector in the characterization of viral mutants in vivo and in vitro is discussed.
范德·韦夫等人（S. 范德·韦夫、J. 布拉德利、E. 维默、F. W. 斯图迪尔和J. 邓恩，《美国国家科学院院刊》83:2330 - 2334，1986年）已经证明，T7 RNA聚合酶在体外合成脊髓灰质炎病毒RNA会产生有感染性的RNA分子；然而，这些分子的感染性仅为从病毒粒子中分离出的RNA的5%。构建了一个质粒T7D - 脊髓灰质炎，它能在体外合成在5'端带有两个额外鸟嘌呤残基的全长RNA分子。然而，T7D - 脊髓灰质炎与范德·韦夫等人构建的质粒不同之处在于，从T7D - 脊髓灰质炎转录的RNA具有真实的3'端，仅以聚腺苷酸核苷酸序列结尾。将这些RNA分子转染到哺乳动物细胞中产生了野生型脊髓灰质炎病毒，其效率与病毒粒子RNA相似。本文讨论了该载体在体内和体外病毒突变体表征中的应用。
