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首页> 外文期刊>Molecular and Cellular Biology >Isolation and characterization of human DNA fragments with nucleotide sequence homologies with the simian virus 40 regulatory region.
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Isolation and characterization of human DNA fragments with nucleotide sequence homologies with the simian virus 40 regulatory region.

机译：具有与猿猴病毒40调控区的核苷酸序列同源性的人DNA片段的分离和表征。

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A recombinant library of human DNA sequences was screened with a segment of simian virus 40 (SV40) DNA that spans the viral origin of replication. One hundred and fifty phage were isolated that hybridized to this probe. Restriction enzyme and hybridization analyses indicated that these sequences were partially homologous to one another. Direct DNA sequencing of two such SV40-hybridizing segments indicated that this was not a highly conserved family of sequences, but rather a set of DNA fragments that contained repetitive regions of high guanine plus cytosine content. These sequences were not members of the previously described Alu family of repeats and hybridized to SV40 DNA more strongly than do Alu family members. Computer analyses showed that the human DNA segments contained multiple homologies with sequences throughout the SV40 origin region, although sequences on the late side of the viral origin contained the strongest cross-hybridizing sequences. Because of the number and complexity of the matches detected, we could not determine unambiguously which of the many possible heteroduplexes between these DNAs was thermodynamically most favored. No hybridization of these human DNA sequences to any other segment of the SV40 genome was detected. In contrast, the human DNA segments isolated cross-hybridized with many sequences within the human genome. We tested for the presence of several functional domains on two of these human DNA fragments. One SV40-hybridizing fragment, SVCR29, contained a sequence which enhanced the efficiency of thymidine kinase transformation in human cells by approximately 20-fold. This effect was seen in an orientation-independent manner when the sequence was present at the 3' end of the chicken thymidine kinase gene. We propose that this segment of DNA contains a sequence analogous to the 72-base-pair repeats of SV40. The existence of such an "activator" element in cellular DNA raises the possibility that families of these sequences may exist in the mammalian genome.
机译：用跨越病毒复制起点的猿猴病毒40（SV40）DNA片段筛选人类DNA序列的重组文库。分离出一百五十个与该探针杂交的噬菌体。限制性酶和杂交分析表明，这些序列彼此部分同源。对两个这样的SV40杂交片段进行直接DNA测序表明，这不是一个高度保守的序列家族，而是一组包含高鸟嘌呤和胞嘧啶含量的重复区域的DNA片段。这些序列不是先前描述的Alu家族重复序列的成员，并且比Alu家族成员更牢固地与SV40 DNA杂交。计算机分析表明，尽管病毒起源后期的序列包含最强的交叉杂交序列，但人类DNA片段在整个SV40起源区域均具有多种同源性，且具有序列。由于检测到的匹配的数量和复杂性，我们无法明确确定在热力学上最喜欢这些DNA之间的许多可能的异源双链体中的哪一个。没有检测到这些人类DNA序列与SV40基因组的任何其他片段的杂交。相反，分离的人类DNA片段与人类基因组中的许多序列交叉杂交。我们测试了其中两个人类DNA片段上几个功能域的存在。一个SV40杂交片段SVCR29包含一个序列，该序列可将人细胞中胸苷激酶的转化效率提高约20倍。当该序列存在于鸡胸苷激酶基因的3'末端时，以定向无关的方式观察到了这种作用。我们建议DNA的此段包含类似于SV40的72个碱基对重复的序列。细胞DNA中这种“活化剂”元件的存在增加了这些序列的家族可能存在于哺乳动物基因组中的可能性。

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