Abstract
Identifying genes that interact with essential genes offers profound theoretical insights into the basic cellular processes of life and practical benefits for fields such as synthetic biology and the fight against drug resistance. Despite its importance, this task is challenging due to the difficulties in creating essential-gene deletion mutants and linking essential gene functions with their interacting partners. In our study, we changed the transformation system to create viable mutants with deletions in genes previously identified as essential in Streptococcus sanguinis . Fortunately, we obtained dozens of essential gene knockout mutants for the first time. These genes, involved in various pathways, are conserved as core components of the essential genome across many bacterial species. Furthermore, through the experimental evolution of 243 independent populations with deletions in these genes, we identified over 1,000 spontaneous suppressor mutations that affect gene essentiality. This collection of spontaneous suppressor mutations, combined with our essential gene deletion mutants, provides a crucial foundation for elucidating gene essentiality and the interactions of essential genes within cellular networks.
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