Camacho E M, Casadesús J
Departamento de Genética, Universidad de Sevilla, Seville 41080, Spain.
Genetics. 2001 Feb;157(2):491-502. doi: 10.1093/genetics/157.2.491.
MudP and MudQ elements were used to induce duplications in Salmonella enterica by formation of a triple crossover between two transduced fragments and the host chromosome. The large size (36 kb) of MudP and MudQ is a favorable trait for duplication formation, probably because homology length is a limiting factor for the central crossover. Additional requirements are a multiplicity of infection of 2 or higher in the infecting phage suspensions (which reflects the need of two transduced fragments) and an exponentially growing recipient (which reflects the need of a chromosome replication fork). We describe a set of 11 strains of S. enterica, each carrying a chromosomal duplication with known endpoints. The collection covers all the Salmonella chromosome except the terminus. For mapping, a dominant marker (e.g., a transposon insertion in or near the locus to be mapped) is transduced into the 11-strain set. Several transductants from each cross are grown nonselectively, and haploid segregants are scored for the presence of the marker. If all the segregants contain the transduced marker, it maps outside the duplication interval. If the marker is found only in a fraction of the segregants, it maps within the duplicated region.
通过在两个转导片段与宿主染色体之间形成三交换,利用MudP和MudQ元件在肠炎沙门氏菌中诱导重复。MudP和MudQ的大尺寸(36 kb)是重复形成的有利特性,这可能是因为同源长度是中央交换的限制因素。其他要求包括感染噬菌体悬液中的感染复数为2或更高(这反映了两个转导片段的需求)以及指数生长的受体(这反映了染色体复制叉的需求)。我们描述了一组11株肠炎沙门氏菌,每株都携带一个具有已知端点的染色体重复。该集合覆盖了除末端之外的所有沙门氏菌染色体。为了进行定位,将一个显性标记(例如,在要定位的基因座内或附近的转座子插入)转导到这11株菌株中。每个杂交的几个转导子在非选择性条件下生长,并对单倍体分离株进行标记存在情况的评分。如果所有分离株都含有转导标记,则它定位于重复区间之外。如果仅在一部分分离株中发现该标记,则它定位于重复区域内。