Strategies for cloning and manipulating natural and synthetic chromosomes

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• 作者：Bogumil J. Karas (1)
  Yo Suzuki (1)
  Philip D. Weyman (1)
  
  1. Synthetic Biology and Bioenergy Group ; J. Craig Venter Institute ; 4120 Capricorn Lane ; La Jolla ; CA ; 92037 ; USA
  
• 关键词：Saccharomyces cerevisiae ; Escherichia coli ; Bacillus subtilis ; Whole ; genome cloning ; Genome transplantation ; Synthetic genomics
• 刊名：Chromosome Research
• 出版年：2015
• 出版时间：February 2015
• 年：2015
• 卷：23
• 期：1
• 页码：57-68
• 全文大小：595 KB
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• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Cell Biology
  Human Genetics
  Animal Genetics and Genomics
  Plant Genetics and Genomics
  
• 出版者：Springer Netherlands
• ISSN：1573-6849

文摘

Advances in synthetic biology methods to assemble and edit DNA are enabling genome engineering at a previously impracticable scale and scope. The synthesis of the Mycoplasma mycoides genome followed by its transplantation to convert a related cell into M. mycoides has transformed strain engineering. This approach exemplifies the combination of newly emerging chromosome-scale genome editing strategies that can be defined in three main steps: (1) chromosome acquisition into a microbial engineering platform, (2) alteration and improvement of the acquired chromosome, and (3) installation of the modified chromosome into the original or alternative organism. In this review, we outline recent progress in methods for acquiring chromosomes and chromosome-scale DNA molecules in the workhorse organisms Bacillus subtilis, Escherichia coli, and Saccharomyces cerevisiae. We present overviews of important genetic strategies and tools for each of the three organisms, point out their respective strengths and weaknesses, and highlight how the host systems can be used in combination to facilitate chromosome assembly or engineering. Finally, we highlight efforts for the installation of the cloned/altered chromosomes or fragments into the target organism and present remaining challenges in expanding this powerful experimental approach to a wider range of target organisms.