Feb, 2019
Exonuclease III (XthA) enforces in vivo DNA cloning of Escherichia coli to create cohesive ends
Journal of Bacteriology
- ,
- Volume
- 201
- Number
- 5
- Language
- Publishing type
- Research paper (scientific journal)
- DOI
- 10.1128/JB.00660-18
- Publisher
- American Society for Microbiology
Cloning of a DNA fragment into a vector is one of the fundamental techniques in recombinant DNA technology. Recently, an in vitro recombination system for DNA cloning was shown to enable the joining of multiple DNA fragments at once. Interestingly, E. coli potentially assembles multiple linear DNA fragments that are introduced into the cell. Improved protocols for this in vivo cloning have realized a high level of usability, comparable to that by in vitro recombination reactions. However, the mechanism of in vivo cloning is highly controversial. Here, we clarified the fundamental mechanism underlying in vivo cloning and also constructed a strain that was optimized for in vivo cloning. Additionally, we streamlined the procedure of in vivo cloning by using a single microcentrifuge tube.
- Link information
- ID information
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- DOI : 10.1128/JB.00660-18
- ISSN : 0021-9193
- eISSN : 1098-5530