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Rec-Mediated Recombinational Hot Spot Activity in Bacteriophage Lambda. I. Hot Spot Activity Associated with Spi- Deletions and bio Substitutions

机译:噬菌体λ中的Rec-介导的重组热点活动。 I.与Spi缺失和生物替代有关的热点活动

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摘要

In order to survey the distribution along the bacteriophage λ chromosome of Rec-mediated recombination events, crosses are performed using conditions which block essentially all DNA synthesis. One parent is density-labeled and carries a genetic marker in the left terminal λ gene (A), while the other parent is unlabeled and carries a genetic marker in the right terminal λ gene (R). Both parents are deleted for the λ recombination genes int and red, together with other recombination-associated genes, by virtue of either (1) a pure deletion or (2) a bio insertion-deletion. The distribution in a cesium density gradient of the resulting A+R+ recombinant phage reflects the chromosomal distribution of the recombination events which gave rise to those phage.Crosses employing either of two different pure deletion phage strains exhibit recombinational hot spot activity located near the right end of the λ chromosome, between the cI and R genes. This hot spot activity persists when unlimited DNA synthesis is allowed. Crosses employing bio1-substituted phage strains exhibit recombinational hot spot activity located to the right of the middle of the chromosome and to the left of the cI gene. Crosses employing either bio1 or bio69-substituted phage strains indicate that the bio-associated hot spot activity occurs in the presence of DNA synthesis, but is dependent on a functional host recB gene.
机译:为了调查沿着Rec介导的重组事件的噬菌体λ染色体的分布,使用基本上阻断所有DNA合成的条件进行杂交。一个亲本被密度标记并在左末端λ基因(A)中带有遗传标记,而另一亲本未被标记并在右末端λ基因(R)中带有遗传标记。借助(1)纯缺失或(2)生物插入缺失,两个λ重组基因int和red的亲本以及其他与重组相关的基因都被删除。所产生的A + R +重组噬菌体在铯密度梯度中的分布反映了重组事件的染色体分布,从而产生了这些噬菌体。使用两种不同纯缺失噬菌体菌株中的任一个的杂交均显示位于右端附近的重组热点活性位于cI和R基因之间的λ染色体。当无限量的DNA合成被允许时,这种热点活动仍然存在。使用bio1取代的噬菌体菌株的杂交显示出重组热点活性,位于染色体中间右侧和cI基因左侧。使用bio1或bio69取代的噬菌体菌株进行的杂交表明,与生物相关的热点活性在存在DNA合成的情况下发生,但取决于功能性宿主recB基因。

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