A novel plasmid-mediated DNA restriction-modification system in E. coli

Plasmid ◽  
1980 ◽  
Vol 4 (3) ◽  
pp. 350-351 ◽  
Author(s):  
L.I. Glatman ◽  
A.F. Moroz ◽  
M.B. Yablokova ◽  
B.A. Rebentish ◽  
G.V. Kc̵holmina
Gene ◽  
1996 ◽  
Vol 183 (1-2) ◽  
pp. 215-218 ◽  
Author(s):  
Richard D. Morgan ◽  
Russell R. Camp ◽  
Geoffrey G. Wilson ◽  
Shuang-yong Xu

Archaea ◽  
2011 ◽  
Vol 2011 ◽  
pp. 1-4 ◽  
Author(s):  
Angel Angelov ◽  
Jörn Voss ◽  
Wolfgang Liebl

Picrophilus oshimaeandPicrophilus torridusare free-living, moderately thermophilic and acidophilic organisms from the lineage ofEuryarchaeota. With a pH optimum of growth at pH 0.7 and the ability to even withstand molar concentrations of sulphuric acid, these organisms represent the most extreme acidophiles known. So far, nothing is known about plasmid biology in these hyperacidophiles. Also, there are no genetic tools available for this genus. We have mobilized the 7.6 Kbp plasmid fromP. oshimaeinE. coliby introducing origin-containing transposons and described the plasmid in terms of its nucleotide sequence, copy number in the native host, mode of replication, and transcriptional start sites of the encoded ORFs. Plasmid pPO1 may encode a restriction/modification system in addition to its replication functions. The information gained from the pPO1 plasmid may prove useful in developing a cloning system for this group of extreme acidophiles.


2021 ◽  
Author(s):  
Stephanie L Brumwell ◽  
Katherine D Van Belois ◽  
Daniel J Giguere ◽  
David R Edgell ◽  
Bogumil J Karas

D. radiodurans has become an attractive microbial platform for the study of extremophile biology and industrial bioproduction. To improve the genomic manipulation and tractability of this species, the development of tools for whole genome engineering and design is necessary. Here, we report the development of a simple and robust conjugation-based transformation system from E. coli to D. radiodurans allowing for the introduction of stable, replicating plasmids expressing antibiotic resistance markers. Using this method with nonreplicating plasmids, we developed a protocol for creating sequential gene deletions in D. radiodurans by target-ing restriction-modification system genes. Importantly, we demonstrated a conjugation-based method for cloning the large (178 kb), high G+C content MP1 megaplasmid from D. radiodurans in E. coli. The conjugation-based tools described here will facili-tate the development of D. radiodurans strains with synthetic genomes for biological studies and industrial applications.


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