Designed horizontal transfer of stable giant DNA released from Escherichia coli.
DNA in the environment is a source to mediate horizontal gene transfer (HGT). Present molecular cloning methods are based on this HGT principle. However, DNA in the extracellular environment, particularly with high molecular-weight, is thought to be prone to shearing or digestion by nucleases. Here we discovered that extracellular plasmid DNA released from lysed Escherichia coli remained intact and stable. Furthermore, it was demonstrated that plasmids up to 100 kb in size were taken up by co-present competent Bacillus subtilis cells. The detailed kinetics of the process together with sensitivity to added DNase I indicated that plasmid DNA released from lysed E. coli into the culture medium was stable enough for quantitative efficacy in the transformation of B. subtilis. Our results will be useful for the development of methods to transfer giant DNAs from general host E. coli without their biochemical purification.
Kaneko
Shinya
S
Mitsubishi Kagaku Institute of Life Sciences, 11 Minamiooya, Machida, Tokyo 194-8511, Japan.
Itaya
Mitsuhiro
M
D001412:Bacillus subtilis / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods*; D003470:Culture Media; D004269:DNA, Bacterial / Q000235:genetics; D003850:Deoxyribonuclease I / Q000494:pharmacology; D004926:Escherichia coli / Q000235:genetics*; D022761:Gene Transfer, Horizontal; D016680:Genome, Bacterial; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D014169:Transformation, Bacterial
D003470:Culture Media; D004269:DNA, Bacterial; D003850:Deoxyribonuclease I