Abstract:
:Top-down reduction of the bacterial genome to construct desired chassis cells is important for synthetic biology. However, the current progress in the field of genome reduction is greatly hindered by indispensable life-essential genes that are interspersed throughout the chromosomal loci. Here, we described a new method designated as "MEGA (Multiple Essential Genes Assembling) deletion and replacement" that functions by assembling multiple essential genes in an E. coli-S. cerevisiae shuttle vector, removing targeted chromosomal regions containing essential and nonessential genes using a one-round deletion, and then integrating the cloned essential genes into the in situ chromosomal loci via I-SceI endonuclease cleavage. As a proof of concept, we separately generated three large deletions (80-205 kbp) in the E. coli MDS42 chromosome. We believe that the MEGA deletion and replacement method has potential to become widely used in large-scale genome reductions in other sequenced organisms in addition to E. coli.
journal_name
ACS Synth Bioljournal_title
ACS synthetic biologyauthors
Xue X,Wang T,Jiang P,Shao Y,Zhou M,Zhong L,Wu R,Zhou J,Xia H,Zhao G,Qin Zdoi
10.1021/sb500324psubject
Has Abstractpub_date
2015-06-19 00:00:00pages
700-6issue
6issn
2161-5063journal_volume
4pub_type
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journal_title:ACS synthetic biology
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pub_type: 信件
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