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大肠杆菌DNA复制蛋白参与噬菌体λ Red介导的同源重组

Involvement of Escherichia coli DNA Replication Proteins in Phage Lambda Red-Mediated Homologous Recombination.

作者信息

Poteete Anthony R

机构信息

Department of Microbiology and Physiological Systems, University of Massachusetts Medical School, Worcester, Massachusetts, United States of America.

出版信息

PLoS One. 2013 Jun 19;8(6):e67440. doi: 10.1371/journal.pone.0067440. Print 2013.

Abstract

The Red recombination system of bacteriophage lambda is widely used for genetic engineering because of its ability to promote recombination between bacterial chromosomes or plasmids and linear DNA species introduced by electroporation. The process is known to be intimately tied to replication, but the cellular functions which participate with Red in this process are largely unknown. Here two such functions are identified: the GrpE-DnaK-DnaJ chaperone system, and DNA polymerase I. Mutations in either function are found to decrease the efficiency of Red recombination. grpE and dnaJ mutations which greatly decrease Red recombination with electroporated DNA species have only small effects on Red-mediated transduction. This recombination event specificity suggests that the involvement of GrpE-DnaJ-DnaK is not simply an effect on Red structure or stability.

摘要

由于噬菌体λ的Red重组系统能够促进细菌染色体或质粒与通过电穿孔导入的线性DNA分子之间的重组,因此它在基因工程中得到了广泛应用。已知该过程与复制密切相关,但在此过程中与Red共同参与的细胞功能在很大程度上尚不清楚。在这里,我们鉴定出了两种这样的功能:GrpE-DnaK-DnaJ伴侣系统和DNA聚合酶I。发现任一功能的突变都会降低Red重组的效率。grpE和dnaJ突变极大地降低了与电穿孔DNA分子的Red重组效率,但对Red介导的转导只有很小的影响。这种重组事件特异性表明,GrpE-DnaJ-DnaK的参与并非仅仅是对Red结构或稳定性的影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bcbe/3686724/0315e6f0a7d1/pone.0067440.g001.jpg

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