RRC ID |
70265
|
Author |
Itaya M, Nagasaku M, Shimada T, Ohtani N, Shiwa Y, Yoshikawa H, Kaneko S, Tomita M, Sato M.
|
Title |
Stable and efficient delivery of DNA to Bacillus subtilis (natto) using pLS20 conjugational transfer plasmids.
|
Journal |
FEMS Microbiol Lett
|
Abstract |
Bacillus subtilis (natto) is generally regarded as a safe bacterium and used as a host for the production of several materials. However, genetic engineering of B. subtilis (natto) is not well established because of poor DNA delivery methods and the lack of a standard strain for the aim. Here, we developed a genetic delivery tool in B. subtilis (natto) using the pLS20 conjugational plasmid (65 kbp). Transmission of pLS20 from B. subtilis 168 to wild-type B. subtilis (natto) did not occur via established mating protocols. We isolated B. subtilis (natto) mutants showing dramatically increased recipient activity. Whole-genome sequence analyses revealed three common alterations: mutations in the restriction endonuclease gene and in the methyl-accepting chemotaxis protein gene, and a 43-kbp deletion at the genome replication termination locus. A representative strain named NEST116 was generated as the first B. subtilis (natto) strain suitable for exploring pLS20-based genetic engineering.
|
Volume |
366(4)
|
Published |
2019-2-1
|
DOI |
10.1093/femsle/fnz032
|
PII |
5307882
|
PMID |
30726909
|
MeSH |
Bacillus subtilis / genetics*
Conjugation, Genetic
DNA, Bacterial / genetics*
Gene Transfer Techniques*
Genetic Engineering / methods*
Mutation
Plasmids / genetics*
|
IF |
1.987
|
Resource |
Prokaryotes B. subtilis |
MBS937
MBS938
MBS939
MBS940
MBS941
MBS942
MBS943 |