Open AccessEffect of B acillus subtilis BsuM restriction–modification on plasmid transfer by polyethylene glycol‐induced protoplast fusion
Author(s) -
Maehara Tomoko,
Itaya Mitsuhiro,
Ogura Mitsuo,
Tanaka Teruo
Publication year - 2011
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2011.02410.x
Subject(s) - bacillus subtilis , protoplast , polyethylene glycol , plasmid , chemistry , bacillales , bacillaceae , microbiology and biotechnology , biology , bacteria , dna , biochemistry , genetics
Polyethylene glycol ( PEG )‐induced cell fusion is a promising method to transfer larger DNA from one cell to another than conventional genetic DNA transfer systems. The laboratory strain B acillus subtilis 168 contains a restriction ( R ) and modification ( M ) system, BsuM , which recognizes the sequence 5′‐ CTCGAG ‐3′. To study whether the BsuM system affects DNA transfer by the PEG ‐induced cell fusion between R + M + and R − M − strains, we examined transfer of plasmids pHV 33 and pLS 32neo carrying no and eight BsuM sites, respectively. It was shown that although the transfer of pLS 32neo but not pHV 33 from the R − M − to R + M + cells was severely restricted, significant levels of transfer of both plasmids from the R + M + to R − M − cells were observed. The latter result shows that the chromosomal DNA in the R − M − cell used as the recipient partially survived restriction from the donor R + M + cell, indicating that the BsuM R − M − strain is useful as a host for accepting DNA from cells carrying a restriction system(s). Two such examples were manifested for plasmid transfer from B acillus circulans and B acillus stearothermophilus strains to a BsuM ‐deficient mutant, B . subtilis RM 125.