MEGA (Multiple Essential Genes Assembling) deletion and replacement method for genome reduction in Escherichia coli.

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 Biol

journal_title

ACS synthetic biology

authors

Xue X,Wang T,Jiang P,Shao Y,Zhou M,Zhong L,Wu R,Zhou J,Xia H,Zhao G,Qin Z

doi

10.1021/sb500324p

subject

Has Abstract

pub_date

2015-06-19 00:00:00

pages

700-6

issue

6

issn

2161-5063

journal_volume

4

pub_type

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