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Further Studies on Bacteriophage T4 DNA Synthesis in Sucrose-Plasmolyzed Cells

机译:进一步研究蔗糖 - 等离子体分解细胞中的噬菌体T4 DNA合成

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This paper describes several technical improvements in the sucrose-plasmolyzed cell system used in earlier experiments on DNA synthesis in situ with Escherichia coli infected by DNA-defective mutants of bacteriophage T4 (W. L. Collinsworth and C. K. Mathews, J. Virol. >13:908-915, 1974). Using this system, which is based primarily on that of M. G. Wovcha et al. (Proc. Natl. Acad. Sci. U.S.A. >70:2196-2200, 1973), we reinvestigated the properties of mutants bearing lesions in genes 1, 41, and 62, and we resolved some disagreements with data reported from that laboratory. We also asked whether the DNA-delay phenotype of T4 mutants is related to possible early leakage of DNA precursors from infected cells. Such cells display defective DNA synthesis in situ, even when ample DNA precursors are made available. Thus, the lesions associated with these mutations seem to manifest themselves at the level of macromolecular metabolism. Similarly, we examined an E. coli mutant defective in its ability to support T4 production, apparently because of a lesion affecting DNA synthesis (L. Simon et al., Nature [London] >252:451-455). In the plasmolyzed cell system, reduced nucleotide incorporation is seen, indicating also that the genetic defect does not involve DNA precursor synthesis. The plasmolyzed cell system incorporates deoxynucleotide 5′-monophosphates into DNA severalfold more rapidly than the corresponding 5′-triphosphates. This is consistent with the idea that DNA precursor-synthesizing enzymes are functionally organized to shuttle substrates to their sites of utilization.
机译:本文介绍了早期使用噬菌体T4的DNA缺陷突变体的DNA合成的DNA合成实验中使用的蔗糖 - 等离子体分解细胞系统的几种技术改进(WL Collinsworth和CK Mathews,J. virol。> 13 :908-915,1974)。使用该系统,主要基于M. G. Wovcha等人。 (Proc。Natl。Acad。SCI。美国 70 :2196-200,1973),我们加入了突变体患者病变的特性,在基因1,41和62中,我们解决了一些与数据分歧的分歧从该实验室报道。我们还询问T4突变体的DNA延迟表型是否与感染细胞的DNA前体的可能早期泄漏有关。这种细胞在原位上显示出缺陷的DNA合成,即使在提供充足的DNA前体是否可用。因此,与这些突变相关的病变似乎在大分子代谢的水平下表现出来。同样,我们检查了一个 e。 Coli 突变体在其支持T4生产的能力中有缺陷,显然是因为影响DNA合成的病变(L.Imon等,Nature [伦敦] > 252 :451-455)。在等离离子体解压缩的细胞体系中,可以看到降低的核苷酸掺入,表明遗传缺陷也不涉及DNA前体合成。等离子体解压缩的细胞系统将脱氧核苷酸5'-单磷酸盐掺入DNA几馏分的比对相应的5'-三磷酸相当。这与DNA前体合成酶在功能上组织成穿梭基板的观点一致,以与其利用率的梭藻。

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