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奇异变形杆菌中由杂交质粒介导的染色体转移

Chromosome transfer in Proteus mirabilis mediated by hybrid plasmid.

作者信息

Coetzee J N

出版信息

J Gen Microbiol. 1975 Jan;86(1):133-46. doi: 10.1099/00221287-86-1-133.

Abstract

A previously-described fused plasmid, P-lacRIdrd19, was found to mediate chromosomal transfer between cells of Proteus mirabilis strain PM5006; PM5006-(P-lacRIdrd19) was usually the donor and various auxotrophs of PM5006 resistant to nalidixic acid and/or streptomycin were recipients. The donor was usually counterselected with nalidixic acid and/or high concentrations of streptomycin. Recombination experiments with single markers indicated a 40-fold variation in recombination frequencies for different markers. Mapping double-auxotrophic markers by their gradient of transmission confirmed this variation and placed each of two independent isolates of eight markers in a linkage group his-ser-ura-pyrB-trp-cys-ade-ilv. Some donor markers did not register. Despite low recombination frequencies, interrupted mating experiments showed a polarity of early marker transfer. The segregation of unselected markers confirmed the order of some markers and showed that genetic material passed from the presumptive donor to the recipient. Recipients with two auxotrophic markers which could not be cotransduced by phage 5006M were converted to prototrophy by conjugation. The plasmid transferred to recipients at high frequency and all recombinants carried it. Recombinants could act as donors in further matings. Recombinants were fully susceptible to phage 5006M, unlike transductants of PM5006 by this phage. Direct involvement of the plasmid was indicated by drastically diminished recombination frequencies in crosses with recipients carrying P-lac as resident. P-lac had previously been shown to reduce the frequency of transfer of the hybrid plasmid to cells harbouring it. The histidine region was the first to register in recipients and recombined at the highest frequency of 5 times 10-6/donor cell. Some temporary association of plasmid and perhaps only the histidine region of the chromosome is favoured as the mechanism of chromosomal transfer. This could explain why not all donor markers could be mapped. Transduction and transformation were excluded as the cause of results.

摘要

一种先前描述的融合质粒P-lacRIdrd19被发现可介导奇异变形杆菌PM5006菌株细胞间的染色体转移;PM5006-(P-lacRIdrd19)通常作为供体,而对萘啶酸和/或链霉素耐药的PM5006的各种营养缺陷型作为受体。供体通常用萘啶酸和/或高浓度链霉素进行反选择。单标记的重组实验表明,不同标记的重组频率有40倍的差异。通过双营养缺陷型标记的传递梯度进行定位证实了这种差异,并将八个标记中的两个独立分离株分别置于一个连锁群his-ser-ura-pyrB-trp-cys-ade-ilv中。一些供体标记未被记录。尽管重组频率较低,但中断杂交实验显示了早期标记转移的极性。未选择标记的分离证实了一些标记的顺序,并表明遗传物质从假定的供体传递到了受体。具有两个不能被噬菌体5006M共转导的营养缺陷型标记的受体通过接合转化为原养型。质粒以高频转移到受体中,所有重组体都携带该质粒。重组体在进一步杂交中可作为供体。与通过该噬菌体转导的PM5006转导子不同,重组体对噬菌体5006M完全敏感。与携带P-lac作为常驻质粒的受体杂交时,重组频率大幅降低,表明质粒直接参与其中。P-lac先前已被证明可降低杂交质粒向携带它的细胞的转移频率。组氨酸区域是第一个在受体中被记录的,并且以最高频率5×10⁻⁶/供体细胞进行重组。质粒与可能仅与染色体的组氨酸区域的某种暂时关联被认为是染色体转移的机制。这可以解释为什么并非所有供体标记都能被定位。转导和转化被排除为结果的原因。

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