Abstract
The use of Bacillus subtilis 168 as the initial host for molecular cloning and subsequent delivery of the engineered DNA to other Bacillus hosts appears attractive, and would lead to an efficient DNA manipulation system. However, methods of delivery to other Bacillus species are limited due to their inability to develop natural competence. An alternative, unexplored conjugational transfer method drew our attention and a B. subtilis native plasmid, pLS30, isolated from B. subtilis (natto) strain IAM1168 was characterized for this aim. The nucleotide sequence (6,610 bp) contained the mob gene and its recognition sequence, oriT, that features pLS30 as a mobile plasmid between Bacillus species on conjugational transfer. Plasmid pLS3001, a chimera with a pBR322-based plasmid prepared in Escherichia coli to confer an antibiotic resistance marker, showed apparent mobilizing activity in the pLS20-mediated conjugational transfer system recently established. The rep gene and associated palT1-like sequence common to all other pLS plasmids previously sequenced indicated that pLS30 is a typical rolling circle replicating (RCR) type plasmid. Due to the significant stability of pLS30 in IAM1168, application of a mobile plasmid would allow quick propagation to Bacillus species.
| Original language | English |
|---|---|
| Pages (from-to) | 557-561 |
| Number of pages | 5 |
| Journal | Journal of Biochemistry |
| Volume | 139 |
| Issue number | 3 |
| DOIs | |
| Publication status | Published - 2006 Mar |
| Externally published | Yes |
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Keywords
- Bacillus natto
- Conjugational transfer
- Plasmid
- Rolling circle replication
- Transformation
ASJC Scopus subject areas
- Biochemistry
Cite this
Experimental basis for a stable plasmid, pLS30, to shuttle between Bacillus subtilis species by conjugational transfer. / Sakaya, Nagayoshi; Kaneko, Shinya; Matsunaga, Satoko; Itaya, Mitsuhiro.
In: Journal of Biochemistry, Vol. 139, No. 3, 03.2006, p. 557-561.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Experimental basis for a stable plasmid, pLS30, to shuttle between Bacillus subtilis species by conjugational transfer
AU - Sakaya, Nagayoshi
AU - Kaneko, Shinya
AU - Matsunaga, Satoko
AU - Itaya, Mitsuhiro
PY - 2006/3
Y1 - 2006/3
N2 - The use of Bacillus subtilis 168 as the initial host for molecular cloning and subsequent delivery of the engineered DNA to other Bacillus hosts appears attractive, and would lead to an efficient DNA manipulation system. However, methods of delivery to other Bacillus species are limited due to their inability to develop natural competence. An alternative, unexplored conjugational transfer method drew our attention and a B. subtilis native plasmid, pLS30, isolated from B. subtilis (natto) strain IAM1168 was characterized for this aim. The nucleotide sequence (6,610 bp) contained the mob gene and its recognition sequence, oriT, that features pLS30 as a mobile plasmid between Bacillus species on conjugational transfer. Plasmid pLS3001, a chimera with a pBR322-based plasmid prepared in Escherichia coli to confer an antibiotic resistance marker, showed apparent mobilizing activity in the pLS20-mediated conjugational transfer system recently established. The rep gene and associated palT1-like sequence common to all other pLS plasmids previously sequenced indicated that pLS30 is a typical rolling circle replicating (RCR) type plasmid. Due to the significant stability of pLS30 in IAM1168, application of a mobile plasmid would allow quick propagation to Bacillus species.
AB - The use of Bacillus subtilis 168 as the initial host for molecular cloning and subsequent delivery of the engineered DNA to other Bacillus hosts appears attractive, and would lead to an efficient DNA manipulation system. However, methods of delivery to other Bacillus species are limited due to their inability to develop natural competence. An alternative, unexplored conjugational transfer method drew our attention and a B. subtilis native plasmid, pLS30, isolated from B. subtilis (natto) strain IAM1168 was characterized for this aim. The nucleotide sequence (6,610 bp) contained the mob gene and its recognition sequence, oriT, that features pLS30 as a mobile plasmid between Bacillus species on conjugational transfer. Plasmid pLS3001, a chimera with a pBR322-based plasmid prepared in Escherichia coli to confer an antibiotic resistance marker, showed apparent mobilizing activity in the pLS20-mediated conjugational transfer system recently established. The rep gene and associated palT1-like sequence common to all other pLS plasmids previously sequenced indicated that pLS30 is a typical rolling circle replicating (RCR) type plasmid. Due to the significant stability of pLS30 in IAM1168, application of a mobile plasmid would allow quick propagation to Bacillus species.
KW - Bacillus natto
KW - Conjugational transfer
KW - Plasmid
KW - Rolling circle replication
KW - Transformation
UR - http://www.scopus.com/inward/record.url?scp=33646375207&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33646375207&partnerID=8YFLogxK
U2 - 10.1093/jb/mvj058
DO - 10.1093/jb/mvj058
M3 - Article
C2 - 16567421
AN - SCOPUS:33646375207
VL - 139
SP - 557
EP - 561
JO - Journal of Biochemistry
JF - Journal of Biochemistry
SN - 0021-924X
IS - 3
ER -