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Alignment of a restriction map with the genetic map of bacteriophage T4.

机译:限制性图谱与噬菌体T4的遗传图谱的比对。

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摘要

A restriction map of the bacteriophage T4 genome was aligned with the T4 genetic map. Included were the cleavage sites for BamHI, BglII, KpnI, PvuI, SalI, and XbaI. The alignment utilized the fact that the T4 genetic map had been oriented previously with respect to a T2/T4 heteroduplex map. DNA fragments from a BglII digestion of cytosine-containing DNA from a T4 dCTPase- denA denB(rIIH23B) alc mutant were hybridized with full-length chromosomal strands of bacteriophage T2, and the heteroduplexes were examined by electron microscopy. From their lengths and patterns of substitution and deletion loops, the heteroduplexes formed with 6 of the 13 BglII fragments could be unambiguously identified and positioned on the T2/T4 heteroduplex map. The ends of the T4 DNA strands in the heteroduplexes directly identified the location of 10 BglII cleavage sites. The remaining three BglII cleavage sites could be assigned to the T2/T4 heteroduplex map based on their relative locations on the restriction map. It was also possible to identify the source of the DNA strands (i.e., T2 or T4) in four previously unassigned deletion loops on the T2/T4 heteroduplex. Among the BglII fragments identified in heteroduplexes was the fragment containing the rIIH23B deletion; this deletion was used as the primary point of reference for alignment of the T4 restriction map with the T2/T4 heteroduplex map and, hence, with the T4 genetic map.
机译:将噬菌体T4基因组的限制性图谱与T4遗传图谱对齐。包括BamHI,BglII,KpnI,PvuI,SalI和XbaI的切割位点。比对利用以下事实:T4遗传图谱先前已相对于T2 / T4异源双链体图进行了定向。将来自T4 dCTPase-denA denB(rIIH23B)alc突变体的含胞嘧啶DNA的BglII消化的DNA片段与噬菌体T2的全长染色体链杂交,并通过电子显微镜检查异源双链体。从它们的长度以及取代和缺失环的模式,可以明确鉴定由13个BglII片段中的6个形成的异源双链体,并将其定位在T2 / T4异源双链体图上。异源双链体中T4 DNA链的末端直接确定了10个BglII切割位点的位置。剩余的三个BglII切割位点可以基于它们在限制图上的相对位置分配给T2 / T4异源双链图。还可以在T2 / T4异源双链体上四个先前未分配的缺失环中鉴定DNA链的来源(即T2或T4)。在异源双链体中鉴定出的BglII片段中有包含rIIH23B缺失的片段。该缺失被用作将T4限制性图谱与T2 / T4异源双链体图谱以及因此与T4遗传图谱进行比对的主要参考点。

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