Shih A, Misra R, Rush M G
Department of Biochemistry, New York University School of Medicine, New York 10016.
J Virol. 1989 Jan;63(1):64-75. doi: 10.1128/JVI.63.1.64-75.1989.
A variety of chemically synthesized oligonucleotides designed on the basis of amino acid and/or nucleotide sequence data were used to detect a large number of novel reverse transcriptase coding sequences in human and mouse DNAs. Procedures involving Southern blotting, library screening, and the polymerase chain reaction were all used to detect such sequences; the polymerase chain reaction was the most rapid and productive approach. In the polymerase chain reaction, oligonucleotide mixtures based on consensus sequence homologies to reverse transcriptase coding sequences and unique oligonucleotides containing perfect homology to the coding sequences of human T-cell leukemia virus types I and II were both effective in amplifying reverse transcriptase-related DNA. It is shown that human DNA contains a wide spectrum of retrovirus-related reverse transcriptase coding sequences, including some that are clearly related to human T-cell leukemia virus types I and II, some that are related to the L-1 family of long interspersed nucleotide sequences, and others that are related to previously described human endogenous proviral DNAs. In addition, human T-cell leukemia virus type I-related sequences appear to be transcribed in both normal human T cells and in a cell line derived from a human teratocarcinoma.
基于氨基酸和/或核苷酸序列数据设计的多种化学合成寡核苷酸被用于检测人和小鼠DNA中大量新的逆转录酶编码序列。涉及Southern印迹、文库筛选和聚合酶链反应的程序都被用于检测此类序列;聚合酶链反应是最快速且高效的方法。在聚合酶链反应中,基于与逆转录酶编码序列的共有序列同源性的寡核苷酸混合物以及与人类I型和II型T细胞白血病病毒编码序列具有完全同源性的独特寡核苷酸,在扩增逆转录酶相关DNA方面均有效。结果表明,人类DNA包含广泛的逆转录病毒相关逆转录酶编码序列,包括一些与人类I型和II型T细胞白血病病毒明显相关的序列、一些与长散在核苷酸序列L-1家族相关的序列,以及其他与先前描述的人类内源性前病毒DNA相关的序列。此外,人类I型T细胞白血病病毒相关序列似乎在正常人类T细胞和源自人畸胎瘤的细胞系中均有转录。