![]() ![]() ![]() ![]() Both managed and wild bee populations are experiencing population declines due to a number of stressors that include viral and bacterial pathogens 4– 6. Honey bees are critical pollinators in agriculture and are essential for global food security 1– 3. alvi genes and to engineer this symbiont for applications including protection of honey bee health. This workflow can be used to study the functions of S. Because it does not require expression of exogenous recombination machinery, SnODIFY is a straightforward, accurate, and lightweight method for genome editing in S. Finally, we confirmed that genome modification is dependent on S. We also used SnODIFY to insert heterologous sequences into the genome by integrating fluorescent protein-coding genes. Mutant phenotypes were validated through knockout of Type 4 pilus genes, which led to reduced biofilm formation. Nearly all transformants had the designed genome modifications, indicating that SnODIFY is highly accurate. We used SnODIFY to create seven single-gene knockout mutants and recovered mutants for all constructs tested. ![]() To solve this problem, we developed a one-step procedure that uses electroporation and homologous recombination, which we term SnODIFY ( Snodgrassella-specific One-step gene Deletion or Insertion to alter Functionalit Y). Despite this importance, there are not currently any easy-to-use methods for modifying the S. The bacterium Snodgrassella alvi is a key member of the bee gut microbiome and has a role in excluding pathogens. As in humans, the health of bees is influenced by their microbiome. Honey bees are economically relevant pollinators experiencing population declines due to a number of threats. ![]()
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