Stable Extracellular DNA: A Novel Substrate for Genetic Engineering that Mimics Horizontal Gene Transfer in Nature

Abstract

AbstractMolecular biology technology generally requires naked DNA purified in a test tube. However, extracellular DNA in the natural environment can contribute to breeding and growth as nutrition or genetic information. Transfer and maintenance of DNA is called horizontal gene transfer (HGT), a mechanism that has certainly played a role during evolution. The naked DNA received by particular microbes through natural transformation processes would almost certainly be extracellular. However, this form of transformation has been rarely studied compared with the other two major mechanisms of HGT: transduction and conjugation.In this chapter, we present our recent discovery that plasmid DNA released from lysed Escherichia coli remains surprisingly stable in the environment. This extracellular DNA, of up to 100 kb and possibly larger, can undergo an HGT-like process into a recipient bacterium capable of natural transformation.This form of DNA transfer, mimicking HGT, could facilitate the delivery of engineered genes without the need for biochemical purification, and permit experimental research into the mechanisms of HGT.KeywordsBacterial Artificial ChromosomeHorizontal Gene TransferNatural TransformationBinary PlasmidSynechocystis PCC6803These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.