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Separation of abnormal cell wall composition from penicillin resistance through genetic transformation of Streptococcus pneumoniae.

机译:通过肺炎链球菌的遗传转化从青霉素抗性中分离异常细胞壁成分。

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摘要

Compared with most penicillin-susceptible isolates of Streptococcus pneumoniae, penicillin-resistant clinical isolate Hun 663 contains mosaic penicillin-binding protein (PBP) genes encoding PBPs with reduced penicillin affinities, anomalous molecular sizes, and also cell walls of unusual chemical composition. Chromosomal DNA prepared from Hun 663 was used to transform susceptible recipient cells to donor level penicillin resistance, and a resistant transformant was used next as the source of DNA in the construction of a second round of penicillin-resistant transformants. The greatly reduced penicillin affinity of the high-molecular-weight PBPs was retained in all transformants through both genetic crosses. On the other hand, PBP pattern and abnormal cell wall composition, both of which are stable, clone-specific properties of strain Hun 663, were changed: individual transformants showed a variety of new, abnormal PBP patterns. Furthermore, while the composition of cell walls resembled that of the DNA donor in the first-round transformants, it became virtually identical to that of susceptible pneumococci in the second-round transformants. The findings indicate that genetic elements encoding the low affinity of PBPs and the penicillin resistance of the bacteria are separable from determinants that are responsible for the abnormal cell wall composition that often accompanies penicillin resistance in clinical strains of pneumococci.
机译:与大多数易感肺炎链球菌的分离株相比,抗青霉素的临床分离株Hun 663含有镶嵌青霉素结合蛋白(PBP)基因,编码的PBP具有降低的青霉素亲和力,异常分子大小以及异常化学组成的细胞壁。由Hun 663制备的染色体DNA用于将易感受体细胞转化为供体水平的青霉素抗性,然后在第二轮青霉素抗性转化子的构建中,将抗性转化子用作DNA的来源。通过两个遗传杂交,在所有转化体中都保留了大分子量PBPs大大降低的青霉素亲和力。另一方面,PBP模式和异常细胞壁组成均稳定,菌株Hun663的克隆特有特性发生了变化:单个转化体显示了各种新的异常PBP模式。此外,尽管第一轮转化子中细胞壁的组成类似于DNA供体的组成,但实际上与第二轮转化子中易感肺炎球菌的细胞壁组成相同。这些发现表明,编码PBPs低亲和力和细菌的青霉素抗性的遗传元件可与导致肺炎链球菌临床菌株中经常伴有青霉素抗性的异常细胞壁组成的决定因素分离。

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