Mutation near the polyoma DNA replication origin permits productive infection of F9 embryonal carcinoma cells
Mutation near the polyoma DNA replication origin permits productive infection of F9 embryonal carcinoma cells
复制标题
多瘤 DNA 复制起点附近的突变允许 F9 胚胎癌细胞的有效感染
DOI：
10.1016/0092-8674(81)90445-1
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发表时间：
1981
期刊：
影响因子：
64.5
通讯作者：
E. Linney
中科院分区：
文献类型：
--
作者：
F. Fujimura;P. Deininger;T. Friedmann;E. Linney
F9小鼠胚胎癌细胞对野生型多瘤病毒的生产性感染具有抗性。对最初感染野生型多瘤病毒的F9细胞继续传代，最终选择出能够对未分化的F9细胞产生感染的多瘤病毒突变体。三个突变体PyFll、PyFll 1和PyF441已被空斑纯化和检测。所有三个PyF突变dna都从Pvu II-4片段的野生型序列中改变，Pvu II-4片段在多瘤基因组上跨越67.6到70.2个图谱单位。PyF441在69.6个图谱单位上有a到G的单碱基变化；PyFl 01和PyFl 11 dna也在69.6个图谱单位上含有这种点突变。此外，PyFl 01和PyFl 11 dna包含点突变的序列分别具有54和31 bp的精确串联重复，并且串联重复的两个拷贝都具有点突变。除了这些变化外，野生型和PyF突变型DNA的核苷酸序列从Bcl I位点的65.6个图谱单位顺时针穿过病毒DNA复制的起源到Bgl I位点的72.2个图谱单位没有差异。双酶切产生的小片段和大片段与限制性内切酶Bcl I和Bgl I的异源组合连接形成的野生型突变型杂交DNA对F9细胞的DNA感染表明，上述DNA序列变化是PyF突变体感染F9细胞能力的原因。
F9 mouse embryonal carcinoma cells are resistant to productive infection by wild-type polyoma virus. Continued passage of F9 cells initially infected with wild-type polyoma virus eventually leads to the selection of polyoma virus mutants that are capable of productive infection of undifferentiated F9 cells. Three mutants, PyFlOl, PyFll 1 and PyF441, have been-plaque-purified and examined. All three PyF mutant DNAs are altered from the wild-type sequence in the Pvu II-4 fragment that spans 67.6 to 70.2 map units on the polyoma genome. PyF441 has a single base change of A to G at 69.6 map units: PyFl 01 and PyFl 11 DNAs also contain this point mutation at 69.6 map units. In addition, PyFl 01 and PyFl 11 DNAs have exact tandem duplications of 54 and 31 bp, respectively, of sequences encompassing the point mutation, and both copies of the tandem duplication have the point mutation. Other than these changes, no difference exists in the nucleotide sequences of wildtype and PyF mutant DNAs from the Bcl I site at 65.6 map units clockwise through the origin of viral DNA replication to the Bgl I site at 72.2 map units. DNA infections of F9 cells with wild-type-mutant hybrid DNAs formed by ligation of heterologous combinations of the small and large DNA fragments generated by double digestion with the restriction enzymes Bcl I and Bgl I show that the DNA sequence changes described above are responsible for the ability of the PyF mutants to infect F9 cells.