Recombineering: A powerful tool for modification of bacteriophage genomes.
Recombineering: A powerful tool for modification of bacteriophage genomes.
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DOI：
10.4161/bact.18778
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发表时间：
2012-01-01
期刊：
影响因子：
--
通讯作者：
Piuri M
中科院分区：
文献类型：
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作者：
Marinelli LJ;Hatfull GF;Piuri M
重组工程是最近开发的一种用于细菌高效遗传操作的技术，它由噬菌体衍生的重组蛋白促进，并且具有使用具有短同源区域的DNA底物的优势。该系统首先在E.大肠杆菌，但后来被改造用于其他细菌。目前已广泛应用于许多不同的系统中，用于各种目的，并已报道了染色体基因敲除、缺失、插入、点突变的构建，以及细菌人工染色体和噬菌体的体内克隆、诱变，以及基因组文库的构建。然而，这些方法也可以有效地应用于噬菌体基因组的遗传修饰，无论是在其原噬菌体和裂解生长状态。不断增长的完全测序的噬菌体集合提出了比它们回答的更多的问题，包括大量基因的未知功能，这些基因没有已知的同源物和未知的功能。噬菌体基因组的改造是解决这些问题的核心，使突变体的简单构建，确定基因的必要性，并阐明基因功能。反过来，我们对噬菌体基因组学的理解的进步应该提供类似的重组工具来解剖许多其他遗传上幼稚的细菌系统。
Recombineering, a recently developed technique for efficient genetic manipulation of bacteria, is facilitated by phage-derived recombination proteins and has the advantage of using DNA substrates with short regions of homology. This system was first developed in E. coli but has since been adapted for use in other bacteria. It is now widely used in a number of different systems for a variety of purposes, and the construction of chromosomal gene knockouts, deletions, insertions, point mutations, as well as in vivo cloning, mutagenesis of bacterial artificial chromosomes and phasmids, and the construction of genomic libraries has been reported. However, these methods also can be effectively applied to the genetic modification of bacteriophage genomes, in both their prophage and lytically growing states. The ever-growing collection of fully sequenced bacteriophages raises more questions than they answer, including the unknown functions of vast numbers of genes with no known homologs and of unknown function. Recombineering of phage genomes is central to addressing these questions, enabling the simple construction of mutants, determination of gene essentiality, and elucidation of gene function. In turn, advances in our understanding of phage genomics should present similar recombineering tools for dissecting a multitude of other genetically naïve bacterial systems.
DOI：
10.4161/bact.18778
发表时间：
2012-01-01
期刊：
Bacteriophage
影响因子：
--
作者：
Marinelli LJ;Hatfull GF;Piuri M
通讯作者：
Piuri M