[严格型和宽松型大肠杆菌不连续培养过程中质粒pBR 322的复制和表达]。

M Hecker, W Wiehle, A Schroeter, F Mach
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引用次数: 5

摘要

携带质粒pBR 322的大肠杆菌CP78和CP79在不连续培养中表现出相似的生长动力学。在氨基酸限制条件下,严格菌株CP78能够合成鸟苷-5′-二磷酸-3′-二磷酸(ppGpp)和鸟苷-5′-三磷酸-3′-二磷酸,而宽松菌株不能产生高度磷酸化的鸟苷核苷酸。在生长的对数阶段,两个菌株含有相似数量的质粒DNA。在氨基酸饥饿期间,质粒DNA仅在松弛菌株中扩增,而在严格菌株中,每个细胞的质粒含量保持不变。在CP78固定相细胞中检测到比CP79细胞更高的质粒编码β -内酰胺酶活性。此外,通过聚丙烯酰胺凝胶电泳分离得到的蛋白组成在两种菌株之间存在显著差异。我们的结果可能表明ppGpp在氨基酸饥饿期间对pBR 322 DNA复制有负性控制。
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[Replication and expression of plasmid pBR 322 during discontinuous culture of a stringent and relaxed Escherichia coli strain].

The E. coli strains CP78 and CP79 carrying the plasmid pBR 322 display similar growth kinetics in discontinuous culture. During limitation of amino acids the stringent strain CP78 is able to synthesize guanosine-5'diphosphate-3'-diphosphate (ppGpp) and guanosine-5'-triphosphate-3' diphosphate, but the relaxed strain can not produce highly phosphorylated guanosine nucleotides. During the logarithmic phase of growth both strains contain similar amounts of plasmid DNA. During amino acid starvation plasmid DNA is amplified in the relaxed strain only, whereas in the stringent strain the plasmid content per cell remains constant. In stationary phase cells of CP78 a higher activity of plasmid-coded beta-lactamase than in CP79 cells was detected. Furthermore, remarkable differences between both strains were observed in the composition of proteins derived from the periplasmic fraction and separated by polyacrylamide gel electrophoresis. Our results might indicate a negative control of pBR 322 DNA replication by ppGpp during amino acid starvation.

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