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噬菌体P22染色体erf区域的遗传分析。

Genetic analysis of the erf region of the bacteriophage P22 chromosome.

作者信息

Fenton A C, Poteete A R

出版信息

Virology. 1984 Apr 15;134(1):148-60. doi: 10.1016/0042-6822(84)90280-0.

Abstract

Derivatives of P22 with deletions of DNA sequences around and including the erf gene were obtained by crossing phages with plasmids containing fragments of the P22 chromosome. In some cases, the parent phages carried a large insertion in sequences not borne by the plasmids. In these cases, deletion of DNA from the phage chromosome to restore terminal repetition (a selectable trait) could be accomplished by recombination between phage and plasmid DNA in chosen sequences flanking the insertion on both sides and borne by the plasmid. In other cases, the parent phages had deletions of a selectable gene, which could be acquired from the plasmid parents only by acquisition of an overlapping deletion. Deletion-bearing P22 strains were tested for growth and homologous genetic recombination in wild-type, recA-, and rec(B or C)- hosts. This analysis indicated the existence of a gene, mapping to the left of erf, that is helpful (but not completely essential) for growth of P22 in a wild-type host. Because P22 lacking this gene grows as well as wild-type P22 on a recBC- host, it has been designated abc (anti-recBC). The abc gene does not appear to be essential for homologous genetic recombination in any host. A plasmid bearing a 1900 base pair fragment of P22 DNA, that expresses erf and abc under the control of the E. coli lac promoter, was constructed. It supports growth and recombination in a recA- host by a phage that lacks all of the genes known to lie between 24 and 9.

摘要

通过将噬菌体与含有P22染色体片段的质粒杂交,获得了P22的衍生物,这些衍生物缺失了包括erf基因及其周围的DNA序列。在某些情况下,亲本噬菌体在质粒未携带的序列中存在大的插入片段。在这些情况下,通过噬菌体与质粒DNA在插入片段两侧侧翼且由质粒携带的特定序列之间的重组,可以实现从噬菌体染色体中删除DNA以恢复末端重复(一种可选择的性状)。在其他情况下,亲本噬菌体缺失了一个可选择的基因,只有通过获得重叠缺失才能从质粒亲本中获得该基因。对携带缺失的P22菌株在野生型、recA - 和rec(B或C) - 宿主中进行生长和同源基因重组测试。该分析表明存在一个基因,定位于erf左侧,它对P22在野生型宿主中的生长有帮助(但不是完全必需)。由于缺乏该基因的P22在recBC - 宿主上的生长与野生型P22一样好,因此将其命名为abc(抗recBC)。abc基因似乎对任何宿主中的同源基因重组都不是必需的。构建了一个携带P22 DNA 1900碱基对片段的质粒,该片段在大肠杆菌lac启动子的控制下表达erf和abc。它支持一个缺乏已知位于24和9之间所有基因的噬菌体在recA - 宿主中的生长和重组。

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