Transposon-Based and Plasmid-Based Genetic Tools for Editing Genomes of Gram-Negative Bacteria
Transposon-Based and Plasmid-Based Genetic Tools for Editing Genomes of Gram-Negative Bacteria
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DOI：
10.1007/978-1-61779-412-4_16
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发表时间：
2011-01-01
期刊：
影响因子：
--
通讯作者：
de Lorenzo, Victor
中科院分区：
文献类型：
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作者：
Martinez-Garcia, Esteban;de Lorenzo, Victor
当代合成生物学议程的很大一部分旨在重新编程微生物以增强现有功能和/或执行新任务。此外，只有在相应的动态系统中进入扰动并监测结果时，才能揭示复杂调节网络甚至单个基因的功能。这些努力依赖于遗传工具的可用性，以成功地按菜单修改目标细菌的染色体。实现这一目标的关键方面包括去除不需要的基因组片段，生产定向突变体和等位基因替代的系统，发现新功能的随机突变文库，以及将更大的遗传网络稳定地植入特定宿主基因组的方法。需要进行这种基因重构操作的革兰氏阴性物种越来越多。然而，除了大肠杆菌之外，可用的分子技术非常有限。在本章中，描述了为促进各种革兰氏阴性微生物的染色体工程程序而量身定制的新工具的使用（例如两种质粒系统：pBAM1和pgm）。
A good part of the contemporary synthetic biology agenda aims at reprogramming microorganisms to enhance existing functions and/or perform new tasks. Moreover, the functioning of complex regulatory networks, or even a single gene, is revealed only when perturbations are entered in the corresponding dynamic systems and the outcome monitored. These endeavors rely on the availability of genetic tools to successfully modify a la carte the chromosome of target bacteria. Key aspects to this end include the removal of undesired genomic segments, systems for the production of directed mutants and allelic replacements, random mutant libraries to discover new functions, and means to stably implant larger genetic networks into the genome of specific hosts. The list of gram-negative species that are appealing for such genetic refactoring operations is growingly expanding. However, the repertoire of available molecular techniques to do so is very limited beyond Escherichia coli. In this chapter, utilization of novel tools is described (exemplified in two plasmids systems: pBAM1 and pEMG) tailored for facilitating chromosomal engineering procedures in a wide variety of gram-negative microorganisms.