Physical mapping of temperature-sensitive mutations of adenoviruses.
Physical mapping of temperature-sensitive mutations of adenoviruses.
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DOI：
10.1016/s0022-2836(75)80046-5
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发表时间：
1975-09
期刊：
影响因子：
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通讯作者：
Joseph Sambrook;Joseph Sambrook;Jim Williams;Jim Williams;Phillip A. Sharp;Phillip A. Sharp;T. Grodzicker;T. Grodzicker
中科院分区：
文献类型：
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作者：
Joseph Sambrook;Joseph Sambrook;Jim Williams;Jim Williams;Phillip A. Sharp;Phillip A. Sharp;T. Grodzicker;T. Grodzicker
分离腺病毒2-猿猴病毒40杂交体的温度敏感性突变体Ad 2 + ND 1 α，并与腺病毒5型（Ad 5）的温度敏感性突变体杂交。筛选出45个野生型重组子，分别用EcoRI、HpaI、BamI、HindIII和SmaI五种限制性内切酶酶切。有15个位点被这些酶切割，这些酶对一个亲本DNA或另一个亲本DNA是独特的。通过凝胶电泳分离通过消化每个重组DNA获得的片段，并与从亲本基因组获得的片段进行比较。发现重组DNA由来自Ad 5和Ad 2 + ND 1的序列组成。知道了5种限制性内切酶切割亲本血清型基因组的位置，就有可能确定每个重组DNA的哪些区域由Ad 5组成，哪些区域由Ad 2 + ND 1序列组成。以这种方式，产生重组体的交换事件被定位，并且突变在亲本基因组中的物理位置被确定。
Temperature-sensitive mutants of the adenovirus 2-simian virus 40 hybrid, Ad2+ND1‡, were isolated and crossed with temperature-sensitive mutants of adenovirus type 5 (Ad5). Forty-five wild type recombinants were selected and their DNAs were separately digested with five restriction endonucleases,EcoRI,HpaI,BamI,HindIII andSmaI. There are 15 sites cleaved by these enzymes that are unique to one parental DNA or the other. The fragments obtained by digestion of each of the recombinant DNAs were separated by gel electrophoresis and compared with those obtained from the parental genomes. The recombinant DNAs were found to consist of sequences derived both from Ad5 and from Ad2+ND1. Knowing the positions at which the five restriction enzymes cleave the genomes of the parental serotypes, it was possible to decide which regions of each recombinant DNA are composed of Ad5 and which of Ad2+ND1 sequences. In this way the cross-over events that generated the recombinants were mapped and the physical locations of thetsmutations in the parental genomes were determined.