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[培养基成分对重组大肠杆菌菌株培养液中细胞外杆菌核糖核酸酶积累的影响]

[Effect of culture medium components on the accumulation of extracellular bacillary ribonucleases in culture fluid of recombinant strains of Escherichia coli].

作者信息

Gabdrakhmanova L A, Znamenskaia L V, Krasnov S I, Chernokal'skaia E B, Leshchinskaia I B

出版信息

Mikrobiologiia. 1996 Sep-Oct;65(5):599-606.

PMID:9102546
Abstract

The effect of the concentrations of peptone, yeast extract, and inorganic phosphate on the expression of genes of extracellular ribonucleases from Bacillus intermedius 7P (binase) and Bacillus amyloliquefaciens H2 (barnase) was studied in Escherichia coli cells transformed with plasmids containing the structural genes of binase or barnase under the control of their own or synthetic regulatory region or the structural binase gene under the control of the regulatory regions of the genes of barnase or Bacillus pumilus RNase. Inorganic phosphate inhibited the expression of the binase gene under the control of its own regulatory region or the regulatory region of the RNase Bp gene. In all other cases, inorganic phosphate produced no effect on the synthesis of RNases by E. coli cells. This difference in the effects of phosphate may be due to the presence of a nucleotide sequence similar to the E. coli Pho box in the promoters of the binase and RNase Bp genes and the absence of this sequence in the barnase gene promoter. It was shown that high peptone and yeast extract concentrations in the cultivation medium are required for good growth of the recombinant E. coli strains and the biosynthesis of RNases.

摘要

研究了蛋白胨、酵母提取物和无机磷酸盐浓度对中间芽孢杆菌7P(binase)和解淀粉芽孢杆菌H2(barnase)细胞外核糖核酸酶基因在大肠杆菌细胞中表达的影响。这些大肠杆菌细胞用含有binase或barnase结构基因的质粒转化,这些基因受其自身或合成调控区控制,或者binase结构基因受barnase或短小芽孢杆菌核糖核酸酶基因调控区控制。无机磷酸盐抑制binase基因在其自身调控区或核糖核酸酶Bp基因调控区控制下的表达。在所有其他情况下,无机磷酸盐对大肠杆菌细胞核糖核酸酶的合成没有影响。磷酸盐作用的这种差异可能是由于binase和核糖核酸酶Bp基因启动子中存在与大肠杆菌Pho盒相似的核苷酸序列,而barnase基因启动子中不存在该序列。结果表明,培养基中高浓度的蛋白胨和酵母提取物是重组大肠杆菌菌株良好生长和核糖核酸酶生物合成所必需的。

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