1887

Abstract

Natural transformation is a mechanism for intra- and interspecific transfer of chromosomal DNA in . During this process a single strand derived from duplex DNA is transported into the cytoplasm and recombined with resident DNA. By electroporation, which introduces duplex DNA into cells, 100-fold lower transformation frequencies of JM300 were observed with shuttle vector or broad-host-range plasmid DNA when the plasmids had replicated in and not in JM300. Moreover, the natural transformation with cloned chromosomal JM300 DNA was reduced about 40-fold when the DNA had not been propagated in JM300 but in . Restriction was also active during natural transformation by single-stranded DNA. Restriction during natural transformation and electroporation was abolished in mutants isolated from mutagenized JM300 cells after applying a multiple plasmid electroporation strategy for the enrichment of restriction-defective strains. The mutants had retained the ability for DNA modification. The strain ATCC 17587 was found to have no restriction–modification system as seen in JM300. It is discussed whether restriction during natural transformation acts at presynaptic or postsynaptic stages of transforming DNA. Restriction as a barrier to transformation apparently contributes to sexual isolation and therefore may promote speciation in the highly diverse species .

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2003-04-01
2020-01-20
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