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[白喉棒状杆菌噬菌体DNA的比较分析及决定白喉毒素合成基因的克隆]

[Comparative analysis of the DNA of Corynebacterium diphtheriae phages and the cloning of the gene determining diphtheria toxin synthesis].

作者信息

Kovgan A A, Zhdanov V M

出版信息

Zh Mikrobiol Epidemiol Immunobiol. 1988 Aug(8):23-31.

PMID:3143205
Abstract

The analysis of the DNA of one nontoxigenic C. diphtheriae phage and two toxigenic ones has revealed that phage phi 984tox+ belongs to omega-like tox+ phages, phage phi 9tox+ is a representative of a new group of phages and phage B (Freeman) tox is a deletion mutant of phage beta. The location of this deletion on the physical map of this phage has been established. To obtain the physical map of phage phi 984tox+, the complete library of internal DNA fragments has been constructed in vector pBR 322. The gene of native diphtheria toxin has been cloned in vectors pBR 322 and pUR 250. Plasmids pUR 250 with the inserts of the toxin gene have been shown to be unstable if tox and lac promoters are located in tandem before the body of the toxin gene. The prolonged cultivation of clones having such structure leads to the formation of a spontaneous mutation located in the region coding the C-end part of the A-fragment of the toxin.

摘要

对一株非产毒型白喉棒状杆菌噬菌体和两株产毒型噬菌体的DNA分析表明,噬菌体φ984tox+属于ω样产毒型噬菌体,噬菌体φ9tox+是一组新噬菌体的代表,而噬菌体B(弗里曼)产毒型是噬菌体β的缺失突变体。已确定该缺失在该噬菌体物理图谱上的位置。为了获得噬菌体φ984tox+的物理图谱,已在载体pBR 322中构建了内部DNA片段的完整文库。天然白喉毒素基因已克隆到载体pBR 322和pUR 250中。如果tox和lac启动子在毒素基因主体之前串联定位,带有毒素基因插入片段的质粒pUR 250已显示不稳定。对具有这种结构的克隆进行长时间培养会导致在编码毒素A片段C末端部分的区域形成自发突变。

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