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通过一条不寻常的途径整合特殊转导噬菌体λcI857 St68 h80 dgnd his,促进缺失的形成并产生一个新的可转位元件。

Integration of specialized transducing bacteriophage lambda cI857 St68 h80 dgnd his by an unusual pathway promotes formation of deletions and generates a new translocatable element.

作者信息

Wolf R E

出版信息

J Bacteriol. 1980 May;142(2):588-602. doi: 10.1128/jb.142.2.588-602.1980.

Abstract

Molecular and genetic studies have revealed that several illegitimate recombinational events are associated with integration of the specialized transducing bacteriophage lambda cI57 St68 h80 dgnd his into either the Escherichia coli chromosome or into a plasmid. Most Gnd+ His+ transductants did not carry the prophage at att phi-80, and 10% were not immune to lambda, i.e., "nonlysogenic." Integration of the phage was independent of the phage Int and Red gene products and of the host's general recombination (Rec) system. In further studies, bacterial strains were selected which carried the phage integrated into an R-factor, pSC50. Restriction endonuclease analysis of plasmid deoxyribonucleic acid (DNA) purified from these strains showed that formation of the hybrid plasmids resulted from recombination between a single region of pSC50 and one of several sites within the lambda-phi 80 portion of the phage. Furthermore the his-gnd region of the phage, present in the chromosome of one nonlysogenic transductant, was shown to be able to translocate to pSC50. Concomitant deletion of phage DNA sequences or pSC50 DNA was frequently observed in conjunction with these integration or translocation events. In supplemental studies, a 22- to 24-megadalton segment of the his-gnd region of the chromosome of a prototrophic recA E. coli strain was shown to translocate to pSC50. One terminus of this translocatable segment was near gnd and was the same as a terminus of the his-gnd segment of the phage which translocated from the chromosome of the nonlysogenic transductant. These data suggest that integration of lambda cI857 St 68 h80 dgnd his may be directed by a recombinationally active sequence on another replicon and that the resulting cointegrate structure is subject to the formation of deletions which extend from the recombinationally active sequence. Translocation of the his-gnd portion of the phage probably requires prior replicon fusion, whereas the his-gnd region of the normal E. coli chromosome may comprise a discrete, transposable element.

摘要

分子和遗传学研究表明,几种异常重组事件与特殊转导噬菌体λcI57 St68 h80 dgnd his整合到大肠杆菌染色体或质粒中有关。大多数Gnd⁺His⁺转导子在att phi-80处不携带原噬菌体,10%对λ不具有免疫性,即“非溶原性”。噬菌体的整合独立于噬菌体Int和Red基因产物以及宿主的一般重组(Rec)系统。在进一步的研究中,选择了携带整合到R因子pSC50中的噬菌体的细菌菌株。对从这些菌株中纯化的质粒脱氧核糖核酸(DNA)进行限制性内切酶分析表明,杂交质粒的形成是由于pSC50的单个区域与噬菌体λ-phi 80部分内的几个位点之一之间的重组。此外,在一个非溶原性转导子的染色体中存在的噬菌体的his-gnd区域被证明能够转移到pSC50。在这些整合或转移事件中经常观察到噬菌体DNA序列或pSC50 DNA的伴随缺失。在补充研究中,一个原养型recA大肠杆菌菌株染色体的his-gnd区域的22至24兆道尔顿片段被证明转移到了pSC50。这个可转移片段的一个末端靠近gnd,与从非溶原性转导子染色体转移来的噬菌体his-gnd片段的一个末端相同。这些数据表明,λcI857 St 68 h80 dgnd his的整合可能由另一个复制子上的重组活性序列引导,并且由此产生的共整合结构容易形成从重组活性序列延伸的缺失。噬菌体his-gnd部分的转移可能需要事先的复制子融合,而正常大肠杆菌染色体的his-gnd区域可能包含一个离散可转座元件。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b67e/294029/d88c757f309c/jbacter00566-0224-a.jpg

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