| 3 | Propagation of Recombinant Genes through Complex Microbiomes with Synthetic Mini-RP4 Plasmid Vectors显示文摘The promiscuous conjugation machinery of the Gram-negative plasmid RP4 has been reassembled in a minimized, highlytransmissible vector for propagating genetically encoded traits through diverse types of naturally occurring microbialcommunities. To this end, the whole of the RP4-encoded transfer determinants (tra, mob genes, and origin of transfer oriT)was excised from their natural context, minimized, and recreated in the form of a streamlined DNA segment borne by anautoselective replicon. The resulting constructs (the pMATING series) could be self-transferred through a variety ofprokaryotic and eukaryotic recipients employing such a rationally designed conjugal delivery device. Insertion of GFP reporterinto pMATING exposed the value of this genetic tool for delivering heterologous genes to both specific mating partners andcomplex consortia (e.g., plant/soil rhizosphere). The results accredited the effective and functional transfer of the recombinantplasmids to a diversity of hosts. Yet the inspection of factors that limit interspecies DNA transfer in such scenarios uncoveredtype VI secretion systems as one of the factual barriers that check otherwise high conjugal frequencies of tested RP4derivatives. We argue that the hereby presented programming of hyperpromiscuous gene transfer can become a phenomenalasset for the propagation of beneficial traits through various scales of the environmental microbiome. | Tomás Aparicio Jillian Silbert Sherezade Cepeda Víctor de Lorenzo | 2022 | BioDesign Research2022，，1： | 0 |
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