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用乳糖启动子替换λdv质粒的pRoR启动子对DNA合成的影响。

Effect of the replacement of pRoR promoter of lambda dv plasmid by lac promoter on the synthesis of DNA.

作者信息

Hase T, Kitamoto A, Masamune Y

出版信息

J Biochem. 1984 Oct;96(4):1205-15. doi: 10.1093/oxfordjournals.jbchem.a134938.

Abstract

Derivatives of lambda dv whose pRoR promoter was replaced by lactose operon (lac promoter) were constructed and cloned in pBR322 plasmid. They were named pLOP-1, 2, and 3, and their structures are shown in Fig. 1. These plasmids were introduced into Escherichia coli (E. coli) lac iq and the effect of the inducer of lac promoter on the synthesis of plasmid DNA was examined. The synthesis of pLOP-2 or 3 DNA was strongly stimulated. pLOP-1, however, responded poorly to the inducer. Plasmids pLOP-2 and 3 were not segregated evenly into daughter cells after the induction and most of the progeny cells did not receive the plasmid. The synthesis of plasmid DNA after induction depended on the function of O and P genes and was inhibited by the addition of rifampicin and chloramphenicol. Most of the plasmid DNA synthesized after the induction cosedimented with the folded host chromosomal complex, suggesting an unusual structure of the DNA. When the inducer was removed, normal segregation of the plasmid resumed and the copy number of plasmid DNA decreased to the original level.

摘要

构建了其pRoR启动子被乳糖操纵子(lac启动子)取代的λdv衍生物,并将其克隆到pBR322质粒中。它们被命名为pLOP-1、2和3,其结构如图1所示。将这些质粒导入大肠杆菌(E. coli)lac iq中,并检测lac启动子诱导剂对质粒DNA合成的影响。pLOP-2或3 DNA的合成受到强烈刺激。然而,pLOP-1对诱导剂反应不佳。诱导后,pLOP-2和3质粒没有均匀地分配到子细胞中,大多数后代细胞没有获得该质粒。诱导后质粒DNA的合成依赖于O和P基因的功能,并受到利福平和平氯霉素添加的抑制。诱导后合成的大多数质粒DNA与折叠的宿主染色体复合物共沉降,表明DNA具有异常结构。当去除诱导剂时,质粒恢复正常分离,质粒DNA的拷贝数降至原始水平。

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