Enhanced shock wave-assisted transformation of Escherichia coli

Abstract

The objective of the study was to demonstrate that shock wave-induced transfer of DNA into bacteria can be increased by enhancing cavitation using dual-pulse (tandem) shock waves. Escherichia coli and plasmid were transferred to test vials. Competent cells were prepared at different concentrations of CaCl(2). Single pulses and tandem shock waves were compared as were three treatment temperatures: 0, 10 and 25 °C. Three delays (250, 500, 750 μs) between double pulses were tested. Characterization was achieved by using a plasmid that provided green fluorescent protein expression. At 0 °C double pulses generated at a delay of 750 μs significantly increased the number of fluorescent colonies compared with single pulses. In general, the lowest temperature enhanced the mean number of transformants compared with the two higher temperatures. A strong influence of the CaCl(2) concentration on the transformation efficiency was also found. The main conclusion is that gene transfer to target cells may be increased up to 50 times at 0 °C by enhancing cavitation using pairs of shock waves.