DNA Molecular Glue Assisted Bacterial Conjugative Transfer

Bacterial conjugation, a commonly used method to horizontally transfer functional genes from donor to recipient strains, plays an important role in the genetic manipulation of bacteria for basic research and industrial production. Successful conjugation depends on the donor-recipient cell recognition and a tight mating junction formation. However, the efficiency of conjugative transfer is usually very low. In this work, we developed a new technique that employed DNA molecule "glue" to increase the match frequency and the interaction stability between the donor and recipient cells. We used two E. coli strains, ETZ and BL21, as a model system, and modified them with the complementary ssDNA oligonucleotides by click chemistry. The conjugation efficiency of the modified bacteria was improved more than 4 times from 10 %-46 %. This technique is simple and generalizable as it only relies on the active amino groups on the bacterial surface. It is expected to have broad applications in constructing engineered bacteria. Bacterial conjugation, a widely used method to transfer DNA from the donor to the recipient strains, shows a very low efficiency with the current method. A new technique that employed DNA molecule "glue" to connect the donor and recipient bacteria to address this issue has been realized for the first time here, mechanistically different from the reported method of using nanomaterials, exploring a new field for the applications of DNA nanotechnology. image