DNA nucleotide sequence heterogeneity between the Towne and AD169 strains of cytomegalovirus
DNA nucleotide sequence heterogeneity between the Towne and AD169 strains of cytomegalovirus
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巨细胞病毒 Towne 株和 AD169 株之间的 DNA 核苷酸序列异质性
DOI：
10.1128/jvi.36.1.152-161.1980
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发表时间：
1980
影响因子：
5.4
通讯作者：
R. Pritchett
中科院分区：
文献类型：
--
作者：
R. Pritchett
DNA-DNA相互重排动力学结果表明，人巨细胞病毒（CMV）Towne株和AD 169株的DNA虽然有90%的核苷酸序列相同，但仍存在约10%的异质性。这意味着，相对于彼此，CMV Towne和CMV AD 169的DNA含有独特的核苷酸序列。为了获得更直接的证据，在存在过量的异源病毒株的未标记DNA的情况下，32 P标记的一个病毒株的DNA重新缔合。在羟基磷灰石柱上从双链DNA中分离那些保留单链的32 P标记的DNA序列，并与含有同源病毒DNA的XbaI限制性酶片段的Southern印迹一起孵育。这种方法不仅富集了每种病毒株特有的核苷酸序列，而且还提供了对其中含有独特序列的限制性内切酶片段的鉴定。在CMV Towne DNA的XbaI片段A、C、G、L、N和Q中发现CMV Towne独特序列。在CMV AD 169 DNA的XbaI片段A、C、G和J中发现CMV AD 169独特序列。这些数据与其他CMV分离株之间的变化可能的意义进行了讨论。
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.