Determination of the chromosomal position effects for plug-and-play application in the Myxococcus xanthus chassis cells

The chromosomal position effect can significantly affect the transgene expression,which may provide an efficient strategy for the inauguration of alien genes in new hosts,but has been less explored rationally.The bacterium Myxococcus xanthus harbors a large circular high-GC genome,and the position effect in this chassis may result in a thousand-fold expression variation of alien natural products.In this study,we conducted transposon insertion at TA sites on the M.xanthus genome,and used enrichment and dilution indexes to respectively appraise high and low expression potentials of alien genes at insertion sites.The enrichment sites are characteristically distributed along the genome,and the dilution sites are overlapped well with the horizontal transfer genes.We experimentally demonstrated the enrichment sites as high expression integration sites(HEISs),and the dilution sites unsuitable for gene integration expression.This work highlights that HEISs are the plug-and-play sites for efficient expression of integrated genes.

Genetic manipulation and tools in myxobacteria for the exploitation of secondary metabolism

Myxobacteria are famous for their capacity for social behavior and natural product biosynthesis.The unique sociality of myxobacteria is not only an intriguing scientific topic but also the main limiting factor for their ma-nipulation.After more than half a century of research,a series of genetic techniques for myxobacteria have been developed,rendering these mysterious bacteria manipulable.Here,we review the advances in genetic manipu-lation of myxobacteria,with a particular focus on the exploitation of secondary metabolism.We emphasize the necessity and urgency of constructing the myxobacterial chassis for synthetic biology research and the exploita-tion of untapped secondary metabolism.