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Alignment of genetic and restriction maps of the photosynthesis region of the Rhodopseudomonas capsulata chromosome by a conjugation-mediated marker rescue technique.

机译:通过缀合介导的标记拯救技术比对荚膜红假单胞菌染色体的光合作用区域的遗传和限制性图谱进行比对。

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摘要

The restriction map of a 46-kilobase fragment of the Rhodopseudomonas capsulata chromosome was aligned with the genetic map of the photosynthesis region of that chromosome by a marker rescue technique. Marker rescue was effected by mobilization of vectors bearing fragments of R. capsulata DNA from Escherichia coli to a set of R. capsulata mutants. Plasmids pDPT51 and pDPT55 were constructed to mediate the intergeneric mobilization of pBR322 derivatives, and a mutant of R. capsulata with improved intergeneric recipient activity was isolated. Four previously unmapped genes affecting bacteriochlorophyll synthesis and two genes affecting photochemical reaction center synthesis have been located by marker rescue. Some of the fragments of R. capsulata DNA are capable of vector-independent complementation, implying that promoters are located on these fragments. Other fragments complement only in one orientation of insertion in the vector, implying transcription from promotors on the vectors and thereby fixing the direction of transcription for those fragments. Still other fragments of DNA show rescue only via recombination between homologous plasmid-borne DNA fragments and chromosomal mutations. The physical dimensions of the genetic map are 3.0 megadaltons per map unit, which agrees with previous estimates based on the size of the R. capsulata gene transfer agent.
机译:通过标记拯救技术,将荚膜红假单胞菌染色体的46个碱基的片段的限制性图谱与该染色体的光合作用区域的遗传图谱对齐。通过将携带来自大肠杆菌的荚膜衣原体DNA片段的载体转移至一组荚膜衣原体突变体来实现标志物拯救。构建质粒pDPT51和pDPT55以介导pBR322衍生物的种间动员,并分离了具有改善的种间受体活性的荚膜红衣菌突变体。通过标记物拯救定位了影响细菌叶绿素合成的四个先前未映射的基因和影响光化学反应中心合成的两个基因。荚膜红球菌DNA的某些片段能够进行不依赖载体的互补作用,这意味着启动子位于这些片段上。其他片段仅在载体插入的一个方向上互补,这意味着从载体上的启动子转录,从而固定了这些片段的转录方向。 DNA的其他片段仅通过同源质粒携带的DNA片段和染色体突变之间的重组而显示出拯救。遗传图谱的物理尺寸为每图谱单位3.0兆道尔顿,这与基于荚膜红衣菌基因转移剂大小的先前估计相符。

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