Pritchett R F
J Virol. 1980 Oct;36(1):152-61. doi: 10.1128/JVI.36.1.152-161.1980.
The results of reciprocal DNA-DNA reassociation kinetics indicated that although the DNAs of human cytomegalovirus (CMV) strains Towne and AD169 shared approximately 90% of their nucleotide sequences, about 10% heterogeneity did exist. The implication was that, with respect to one another, the DNAs of CMV Towne and CMV AD169 contained unique nucleotide sequences. To obtain more direct evidence, 32P-labeled DNA of one virus strain was reassociated in the presence of excess unlabeled DNA of the heterologous virus strain. Those 32P-labeled DNA sequences remaining single stranded were separated from double-stranded DNA on hydroxyapatite columns and incubated with Southern blots containing XbaI restriction enzyme fragments of the homologous virus DNA. This approach not only enriched for nucleotide sequences unique to each strain of virus, but also provided for the identification of the restriction enzyme fragments in which the unique sequences were contained. The CMV Towne unique sequences were found in XbaI fragments A, C, G, L, N, and Q of CMV Towne DNA. The CMV AD169 unique sequences were found in XbaI fragments A, C, G, and J of CMV AD169 DNA. The possible significance of these data with respect to variation among other CMV isolates is discussed.
相互DNA-DNA复性动力学结果表明,尽管人巨细胞病毒(CMV)毒株汤氏株和AD169株的DNA约90%的核苷酸序列相同,但确实存在约10%的异质性。这意味着,CMV汤氏株和CMV AD169株的DNA相互之间含有独特的核苷酸序列。为获得更直接的证据,在异源病毒株的过量未标记DNA存在的情况下,使一种病毒株的32P标记DNA复性。那些保持单链的32P标记DNA序列在羟基磷灰石柱上与双链DNA分离,并与含有同源病毒DNA的XbaI限制性酶切片段的Southern印迹杂交孵育。这种方法不仅富集了每种病毒株特有的核苷酸序列,还能鉴定出含有独特序列的限制性酶切片段。CMV汤氏株的独特序列存在于CMV汤氏株DNA的XbaI片段A、C、G、L、N和Q中。CMV AD169株的独特序列存在于CMV AD169株DNA的XbaI片段A、C、G和J中。本文讨论了这些数据对于其他CMV分离株变异的可能意义。