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Overview
Authored
(1)
CL
Cammie F Lesser
Department of Medicine, Division of Infectious Diseases, Massachusetts General Hospital, USA
Research fields
Microbiology
Peer-reviewed
Preprint
Transfer of Large Contiguous DNA Fragments onto a Low Copy Plasmid or into the Bacterial Chromosome
Authors:
Analise Z Reeves
and
Cammie F Lesser
,
date:
11/20/2016,
view:
10391,
Q&A:
0
Bacterial pathogenicity islands and other contiguous operons can be difficult to clone using conventional methods due to their large size. Here we describe a robust 3-step method to transfer large defined fragments of DNA from virulence plasmids or cosmids onto smaller autonomously replicating plasmids or directly into defined sites in the bacterial chromosome that incorporates endogenous yeast and λ Red homologous recombination systems. This methodology has been successfully used to isolate and integrate at least 31 kb of contiguous DNA and can be readily adapted for the recombineering of
E. coli
and its close relatives.
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