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在具有改变的抗生素抗性水平和/或接合能力的大肠杆菌菌株中R质粒脱氧核糖核酸的体内转录

In vivo transcription of R-plasmid deoxyribonucleic acid in Escherichia coli strains with altered antibiotic resistance levels and/or conjugal proficiency.

作者信息

Davis R, Vapnek D

出版信息

J Bacteriol. 1976 Mar;125(3):1148-55. doi: 10.1128/jb.125.3.1148-1155.1976.

Abstract

The amounts of plasmid deoxyribonucleic acid (DNA) and the levels of the in vivo transcription of the Escherichia coli plasmids R538-1 (repressed for conjugal transfer) and R538-1drd (derepressed for transfer) were determined by DNA-DNA hybridization and DNA-ribonucleic acid hybridization, respectively. The results demonstrate that the level of plasmid transcription is increased by two-fold in the strain carrying the derepressed plasmid, compared to an isogenic strain carrying the repressed plasmid, whereas the amount of plasmid DNA is approximately the same, suggesting that the transfer genes are under transcriptional control. Levels of plasmid DNA, plasmid DNA transcription, and chloramphenicol acetyltransferase activity were also compared in a mutant strain that carried the R538-1drd plasmid and was resistant to high levels of antibiotics. This strain produces about 13 copies of plasmid DNA per chromosome compared to five copies for the parent strain. The level of transcription of plasmid DNA was found to be twofold higher in the high-level resistant strain, whereas the level of chloramphenition, acetyltransferase activity was increased by 10-fold. In addition the levels of plasmid DNA transcription and chloramphenicol acetyltransferase activity in the high-level resistant strain were found to be further increased by the presence of high levels of chloramphenicol in the growth medium. The amount of plasmid DNA remained constant under these conditions, indicating that high levels of chloramphenicol can stimulate the expression of plasmid genes at the level of transcription in this strain.

摘要

分别通过DNA-DNA杂交和DNA-核糖核酸杂交测定了大肠杆菌质粒R538-1(接合转移受抑制)和R538-1drd(转移去抑制)的质粒脱氧核糖核酸(DNA)量及体内转录水平。结果表明,与携带受抑制质粒的同基因菌株相比,携带去抑制质粒的菌株中质粒转录水平提高了两倍,而质粒DNA量大致相同,这表明转移基因处于转录控制之下。还比较了携带R538-1drd质粒且对高水平抗生素具有抗性的突变菌株中的质粒DNA水平、质粒DNA转录水平和氯霉素乙酰转移酶活性。与亲本菌株的五个拷贝相比,该菌株每条染色体产生约13个质粒DNA拷贝。发现高水平抗性菌株中质粒DNA的转录水平高两倍,而氯霉素乙酰转移酶活性提高了10倍。此外,发现生长培养基中高水平氯霉素的存在会进一步提高高水平抗性菌株中的质粒DNA转录水平和氯霉素乙酰转移酶活性。在这些条件下,质粒DNA量保持恒定,表明高水平氯霉素可在该菌株的转录水平上刺激质粒基因的表达。

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