What is it about?
Lactobacilli are promising candidates for developing genetically altered strains with therapeutic and diagnostic capabilities. Creating such strains employs recombinant plasmids that can be generated rapidly for faster screening. Our protocol makes this possible by using the widely known Gibson Assembly method and maintenance of the plasmid directly in the Lactobacillus instead of incompatible intermediate hosts.
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Why is it important?
Our work is unique because it allows the user the flexibility to create recombinant plasmids in the host strain rather than passing it through intermediate hosts, which are highly responsible for generating unwanted mutations in heterologous genes.
Perspectives
This protocol provides additional scope to develop hard-to-develop recombinant plasmids directly in the host strain without significantly losing time and resources.
Sourik Dey
Leibniz Institute for New Materials
Read the Original
This page is a summary of: In vitro assembly of plasmid DNA for direct cloning in Lactiplantibacillus plantarum WCSF1, PLoS ONE, February 2023, PLOS,
DOI: 10.1371/journal.pone.0281625.
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