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噬菌体φX174和G4 DNA的异源双链体:与遗传图谱的定位及与核苷酸序列预测结果的比较

Heteroduplexes of phiX174 and G4 DNAs: orientation to genetic map and comparison with predictions from nucleotide sequences.

作者信息

Smiley B L, Warner R C

出版信息

Nucleic Acids Res. 1979;6(5):1979-91. doi: 10.1093/nar/6.5.1979.

Abstract

Heteroduplexes between the viral DNA of phiX174 and DNA from the replicative form (RF) of phage G4 were examined by electron microscopy. The single Eco RI site of G4-RF was utilized as a physical marker by preparing the heteroduplexes from the denatured, linear DNA obtained by restricting G4-RF with Eco RI endonuclease. Restriction fragments of phiX were used in a separate series of heteroduplexes to align the heteroduplex map and the G4 Eco RI site with the similar genetic maps of the two phages. The positions of the branch migrating junctions of recombinant phiX-G4 figure-8s, previously located only with respect to the G4-Eco RI site, have now been located with high proability within the gene A region of the two genomes. The degree of mismatch between the known nucleotide sequences of phi X and G4 accounts for positions of all of the regions of single-strandedness in the observed heteroduplexes, but unexplained discrepancies were also found.

摘要

通过电子显微镜检查了噬菌体φX174的病毒DNA与噬菌体G4复制型(RF)的DNA之间的异源双链体。通过用Eco RI核酸内切酶切割G4 - RF获得变性线性DNA来制备异源双链体,将G4 - RF的单一Eco RI位点用作物理标记。在另一系列异源双链体中使用φX的限制片段,将异源双链体图谱和G4 Eco RI位点与两个噬菌体的相似遗传图谱对齐。重组φX - G4“8”字形分支迁移连接点的位置,以前仅相对于G4 - Eco RI位点定位,现在已高概率地定位在两个基因组的基因A区域内。φX和G4已知核苷酸序列之间的错配程度解释了观察到的异源双链体中所有单链区域的位置,但也发现了无法解释的差异。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/29ad/327825/09ad73df0005/nar00446-0237-a.jpg

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